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Department of Pharmaceutical Botany, “Iuliu Hațieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania
Department of Pharmacy, “G. d’Annunzio” University of Chieti-Pescara, 66100 Chieti, Italy
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Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods

Abstract submission deadline
closed (31 May 2023)
Manuscript submission deadline
closed (31 July 2023)
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208525

Topic Information

Dear Colleagues,

In recent years, the scientific community has shown great interest in the field of plant antioxidants, and a great number of studies have been published documenting their health-promoting effects as well as their involvement as pharmacological agents in different molecular signaling pathways. A variety of plant materials are known to be natural sources of antioxidants, such as herbs, spices, seeds, fruits, and vegetables. The interest in these natural components is driven not only by their biological value, but also by their economic and ecological impact, as most of them may be extracted from food byproducts, under-exploited plant species, or traditionally used medicinal and edible plants. The current Topic gathers five well-respected journals from MDPI aiming to offer the possibility for researchers to publish their results concerning plant antioxidants in the most suitable journal, featuring great visibility for their research. The Topic welcomes manuscripts regarding any aspects of plant antioxidants in relation to their use in nutrition, medicinal chemistry, pharmacology and pharmacy, and food chemistry and technology.

Dr. Andrei Mocan
Prof. Dr. Simone Carradori 
Topic Editors

 

Keywords

  • natural antioxidants
  • semi-synthetic antioxidant derivatives from natural sources
  • antioxidants in nutrition
  • antioxidant assays
  • radical scavenging agents
  • oxygen reactive species
  • food components
  • food antioxidants
  • thermal-induced modifications in food antioxidants

Participating Journals

Journal Name Impact Factor CiteScore Launched Year First Decision (median) APC
Antioxidants
antioxidants 		6.0 10.6 2012 15.5 Days CHF 2900
Foods
foods 		4.7 7.4 2012 14.3 Days CHF 2900
Molecules
molecules 		4.2 7.4 1996 15.1 Days CHF 2700
Oxygen
oxygen 		- - 2021 23.1 Days CHF 1000
Plants
plants 		4.0 6.5 2012 18.2 Days CHF 2700

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Published Papers (78 papers)

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14 pages, 2397 KiB  
Article
Phenolic Profile and Bioactivity Changes of Lotus Seedpod and Litchi Pericarp Procyanidins: Effect of Probiotic Bacteria Biotransformation
by Junren Wen, Yong Sui, Shuyi Li, Jianbin Shi, Sha Cai, Tian Xiong, Fang Cai, Lei Zhou, Shengnan Zhao and Xin Mei
Antioxidants 2023, 12(11), 1974; https://doi.org/10.3390/antiox12111974 - 7 Nov 2023
Viewed by 1639
Abstract
Theoretically, lactic acid bacteria (LABs) could degrade polyphenols into small molecular compounds. In this study, the biotransformation of lotus seedpod and litchi pericarp procyanidins by Lactobacillus plantarum 90 (Lp90), Streptococcus thermophilus 81 (ST81), Lactobacillus rhamnosus HN001 (HN001), [...] Read more.
Theoretically, lactic acid bacteria (LABs) could degrade polyphenols into small molecular compounds. In this study, the biotransformation of lotus seedpod and litchi pericarp procyanidins by Lactobacillus plantarum 90 (Lp90), Streptococcus thermophilus 81 (ST81), Lactobacillus rhamnosus HN001 (HN001), and Pediococcus pentosus 06 (PP06) were analysed. The growth curve results indicated that procyanidins did not significantly inhibit the proliferation of LABs. Ultra-high-performance liquid chromatography high-resolution mass spectrometry (UPLC–HRMS) revealed that procyanidin B2 and procyanidin B3 in lotus seedpod decreased by 62.85% and 25.45%, respectively, with ST81 metabolised, while kaempferol and syringetin 3-O-glucoside content increased. Although bioconversion did not increase the inhibitory function of procyanidins against glycosylation end-products in vitro, the 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) free radical scavenging capacity and ferric reducing antioxidant power of litchi pericarp procyanidins increased by 157.34% and 6.8%, respectively, after ST81 biotransformation. These findings may inspire further studies of biological metabolism of other polyphenols and their effects on biological activity. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Effects of different concentrations of LSPC and LPPC on LAB growth. (<b>A</b>–<b>D</b>) Kinetics growth of <span class="html-italic">Lp90</span> (<b>A</b>), <span class="html-italic">ST81</span> (<b>B</b>), <span class="html-italic">HN001</span> (<b>C</b>), and <span class="html-italic">PP06</span> (<b>D</b>) in MRS broth with LSPC and LPPC at 37 °C.</p> Full article ">Figure 2
<p>The original procyanidin and LAB metabolite compositions. Representative chromatograms of the identified phenolic compounds in LSPC (<b>A</b>) and LPPC (<b>B</b>). Phenolic compounds identified in LSPC: (S1) procyanidin B3; (S2) (+)-catechin; (S3) procyanidin B2; (S4) rutin; (S5) kaempferol 3-O-glucoside; (S6) myricetin 3-O-glucoside; and (S7) syringetin 3-O-glucoside. Phenolic compounds identified in LPPC: (P1) (+)-catechin; (P2) (−)-epicatechin; (P3) A-type procyanidin trimer; and (P4) procyanidin A2. Content of individual phenolic compounds in LSPC (<b>C</b>) and LPPC (<b>D</b>) at different sampling points (mg), and different lowercase letters indicate significant differences between sampling times by the Tukey test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Inhibition rates of LSPC and LPPC metabolite on fluorescent advanced AGEs. Data shown are mean ± SEM, and different lowercase letters indicate significant differences between metabolites of various LABs by the Tukey test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Antioxidant activity of metabolites. The FRAP, ABTS, and ORAC activity of LSPC (<b>A</b>–<b>C</b>) and LPPC (<b>D</b>–<b>F</b>) metabolites after gastrointestinal digestion in vitro. Data shown are mean ± SEM, and different uppercase letters indicate significant differences between gastric and intestinal chyme, while the lowercase letters indicate significant differences between metabolites of various LABs by the Turkey test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Correlation analysis of LSPC (<b>A</b>) and LPPC (<b>B</b>) metabolites with antioxidant activities determined by diverse assays in vitro.</p> Full article ">
16 pages, 3287 KiB  
Article
Andrographolide Induces ROS-Mediated Cytotoxicity, Lipid Peroxidation, and Compromised Cell Integrity in Saccharomyces cerevisiae
by Tanaporn Phetruen, Bloem van Dam and Sittinan Chanarat
Antioxidants 2023, 12(9), 1765; https://doi.org/10.3390/antiox12091765 - 14 Sep 2023
Cited by 1 | Viewed by 1724
Abstract
Andrographolide, a bioactive compound found in Andrographis paniculata, has gained significant attention for its potential therapeutic properties. Despite its promising benefits, the understanding of its side effects and underlying mechanisms remains limited. Here, we investigated the impact of andrographolide in Saccharomyces cerevisiae [...] Read more.
Andrographolide, a bioactive compound found in Andrographis paniculata, has gained significant attention for its potential therapeutic properties. Despite its promising benefits, the understanding of its side effects and underlying mechanisms remains limited. Here, we investigated the impact of andrographolide in Saccharomyces cerevisiae and observed that andrographolide induced cytotoxicity, particularly when oxidative phosphorylation was active. Furthermore, andrographolide affected various cellular processes, including vacuole fragmentation, endoplasmic reticulum stress, lipid droplet accumulation, reactive oxygen species levels, and compromised cell integrity. Moreover, we unexpectedly observed that andrographolide induced the precipitation of biomolecules secreted from yeast cells, adding an additional source of stress. Overall, this study provides insights into the cellular effects and potential mechanisms of andrographolide in yeast, shedding light on its side effects and underlying cytotoxicity pathways. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Andrographolide exhibits cytotoxicity in <span class="html-italic">S. cerevisiae</span> with ROS synergy. (<b>A</b>) Chemical structure of andrographolide. (<b>B</b>) Growth curves of wild-type (WT) yeast cells in synthetic minimal media supplemented with 2% glycerol in the presence or absence of andrographolide (AD). DMSO serves as control. Means ± SD (error bars) are shown (<span class="html-italic">n</span> = 3). (<b>C</b>,<b>D</b>) Similar to (<b>B</b>), but <span class="html-italic">ccs1Δ</span> and <span class="html-italic">sod2Δ</span> yeast mutant cells were used, respectively.</p> Full article ">Figure 2
<p>Andrographolide induces vacuole fragmentation. (<b>A</b>) Representatives of three vacuole morphologies observed in FM4-64 staining. Scale bar, 5 µm. (<b>B</b>) Quantification of different vacuole morphologies in the presence or absence of andrographolide (AD). DMSO serves as control. Means ± SD (error bars) from two replicates with more than 2000 cells per replicate are shown. Statistical analysis was carried out using a two-way ANOVA with Sidak’s comparison between AD and DMSO serving as control conditions (<span class="html-italic">p</span> = 0.0009 for sole spherical, <span class="html-italic">p</span> = 0.003 for fragmented spherical). (<b>C</b>,<b>D</b>) Similar to (<b>B</b>), but <span class="html-italic">vma4Δ</span> and <span class="html-italic">vma9Δ</span> yeast mutant cells were used, respectively.</p> Full article ">Figure 3
<p>Andrographolide induces ER stress and accumulation of lipid droplets. (<b>A</b>) Splicing status of <span class="html-italic">HAC1</span> mRNA from yeast cells treated with andrographolide for 1 h and assessed by RT-PCR. DTT and DMSO serve as positive and negative controls, respectively. (<b>B</b>) Lipid droplets in cells treated with or without andrographolide for the indicated time period were stained with BODIPY 493/503. Scale bar, 5 µm. (<b>C</b>) Quantification of (<b>B</b>). Each dot represents the fluorescence intensity per cell area measured by ImageJ 1.53t. Statistical analysis was carried out by two-tailed unpaired t-test with a Mann–Whitney test (****, <span class="html-italic">p</span> &lt; 0.0001; ns, not significant).</p> Full article ">Figure 4
<p>Andrographolide induces lipid peroxidation and ROS elevation. (<b>A</b>) Reactive oxidative species (ROS) in cells exposed to andrographolide for the indicated time period were assessed by 2′,7′-dichlorofluorescin diacetate (DCFDA) staining. Each dot represents the fluorescence intensity per 1 mg/mL protein. Error bars indicate the SD of three independent biological replicates. Statistical significance was calculated using a two-way ANOVA with Sidak’s multiple comparisons (***, <span class="html-italic">p</span> &lt; 0.001; ns, not significant). (<b>B</b>) Similar to (<b>A</b>), but ROS-mediated lipid oxidation was assessed by TBARS assay via spectrophotometric detection of its end product malondialdehyde at 535 nm. Each dot represents the amount of calculated MDA in µmol/mg protein unit. Error bars indicate the SD of three independent biological replicates. Statistical significance was calculated using a two-way ANOVA with Sidak’s multiple comparisons (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ns, not significant).</p> Full article ">Figure 5
<p>Andrographolide compromises cell integrity and affects ergosterol-dependent membrane stability. (<b>A</b>) Zymolyase sensitivity of yeast cells treated with andrographolide. Cells were treated with 0.4 U of Zymolyase and OD600 was observed at the indicated time point. DMSO serves as control. (<b>B</b>) Propidium iodide staining of yeast cells treated with andrographolide. DMSO serves as control. Scale bar, 10 µm. (<b>C</b>) Quantification of (<b>B</b>). Means ± SD (error bars) from three replicates (<span class="html-italic">n</span> &gt; 100 per replicate) are shown. Statistical analysis was carried out using a two-way ANOVA with Sidak’s comparison between AD and DMSO serving as control conditions (<span class="html-italic">p</span> = 0.017 for PI positive). (<b>D</b>) Similar to <a href="#antioxidants-12-01765-f001" class="html-fig">Figure 1</a>B, but <span class="html-italic">erg6Δ</span> mutant cells were used.</p> Full article ">Figure 6
<p>Andrographolide induces sheet-like structures, impacting yeast cells. (<b>A</b>) Scanning electron micrographs of yeast cells treated with or without andrographolide. Two representative micrographs are shown. The red arrows indicate sheet-like structures that cover yeast cells. (<b>B</b>) A diagram depicting an andrographolide-induced precipitation assay. Either fresh or cultured media were mixed with andrographolide to the final concentration of 1 mg/mL, centrifuged, and washed with sterile distilled water. (<b>C</b>) The precipitation in SG media from the experiment represented in (<b>B</b>). (<b>D</b>) Similar to (<b>A</b>), but the pellets formed after mixing andrographolide with cultured media were analyzed. (<b>E</b>) The pellets were resuspended in 2× SDS-PAGE loading buffer and subjected to SDS gel electrophoresis followed by silver staining. (<b>F</b>) Quantification of fluorescent intensity from BODIPY 493/503 staining. Each dot represents fluorescence intensity per cell area measured by ImageJ (version 1.53t). Statistical analysis was carried out via a two-tailed unpaired <span class="html-italic">t</span>-test with a Mann–Whitney test (*, <span class="html-italic">p</span> &lt; 0.05; ****, <span class="html-italic">p</span> &lt; 0.0001).</p> Full article ">Figure 7
<p>Model of andrographolide’s mechanism of action in budding yeast <span class="html-italic">S. cerevisiae</span>. (The figure was created at <a href="http://BioRender.com" target="_blank">BioRender.com</a>).</p> Full article ">
22 pages, 5998 KiB  
Article
Optimization of Enzyme−Assisted Aqueous Extraction of Polysaccharide from Acanthopanax senticosus and Comparison of Physicochemical Properties and Bioactivities of Polysaccharides with Different Molecular Weights
by Xueyan Wang, Yuanyuan Su, Jianqing Su, Jiaojiao Xue, Rui Zhang, Xiaoli Li, Ying Li, Yi Ding and Xiuling Chu
Molecules 2023, 28(18), 6585; https://doi.org/10.3390/molecules28186585 - 12 Sep 2023
Cited by 9 | Viewed by 1845
Abstract
To obtain the optimal process for the enzyme−assisted aqueous extraction of polysaccharides from Acanthopanax senticosus, and study the physicochemical properties of polysaccharides of different molecular weights, the extraction of Acanthopanax polysaccharides was optimized using the BBD response surface test. The polysaccharides with [...] Read more.
To obtain the optimal process for the enzyme−assisted aqueous extraction of polysaccharides from Acanthopanax senticosus, and study the physicochemical properties of polysaccharides of different molecular weights, the extraction of Acanthopanax polysaccharides was optimized using the BBD response surface test. The polysaccharides with different molecular weights were obtained by ethanol−graded precipitation at 40%, 60%, and 80%, which were presented as ASPS40, ASPS60, and ASPS80. The polysaccharides were analyzed by HPGPC, ion chromatography, FT−IR, UV, SEM, TGA, XRD, Congo red, and I2−KI tests. The antioxidant assay was used to evaluate their antioxidant properties in vitro. The findings demonstrated that the recovery rate of Acanthopanax polysaccharide was 10.53 ± 0.682%, which is about 2.5 times greater compared to the conventional method of hot water extraction. Based on FT−IR, TGA, polysaccharides with different molecular weights did not differ in their structure or thermal stability. The XRD suggests that the internal structure of ASPSs is amorphous. Congo red and I2−KI showed that all three polysaccharides had triple helix structures with longer branched chains and more side chains. Furthermore, the antioxidant results showed the antioxidant activity of polysaccharides is not only related to the molecular weight size but also can be related to its composition and structure. These studies developed a green, and scalable method to produce polysaccharides from Acanthopanax senticosus and evaluated the properties of Acanthopanax polysaccharides of different molecular weights. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>The effect of each enzyme on the yield of polysaccharides from <span class="html-italic">A. senticosus</span>: (<b>A</b>) the amount of cellulase; (<b>B</b>) the amount of pectinase; (<b>C</b>) the amount of hemicellulase. Note: a, b, and c: represent the significance of the difference within the graph. Containing the same letter means the difference is insignificant (<span class="html-italic">p</span> &gt; 0.05).</p> Full article ">Figure 2
<p>The effect of enzymatic conditions on the extraction rate of polysaccharides from <span class="html-italic">A. senticosus</span>: (<b>A</b>) the amount of enzymes; (<b>B</b>) the pH; (<b>C</b>) the extraction temperature; (<b>D</b>) the extraction time; (<b>E</b>) the solid–liquid ratio. Note: a, b, c and d: represent the significance of the difference within the graph. Containing the same letter means the difference is not significant (<span class="html-italic">p</span> &gt; 0.05).</p> Full article ">Figure 3
<p>The 3D response surface plots and contour plots (<b>A</b>–<b>F</b>) of the influence of interactions of various factors on the extraction rate of ASPS. Note: SLR is the solid–liquid ratio.</p> Full article ">Figure 4
<p>FT−IR images of ASPS−4, ASPS−60, ASPS−80.</p> Full article ">Figure 5
<p>UV spectra of ASPS−40, ASPS−60, ASPS−80.</p> Full article ">Figure 6
<p>TGA (<b>a</b>) and DTG (<b>b</b>) analysis images of ASPS−40, ASPS−60, and ASPS−80.</p> Full article ">Figure 7
<p>SEM images of ASPS−40, ASPS−60, and ASPS−80 (5000×).</p> Full article ">Figure 8
<p>XRD spectra of ASPSs.</p> Full article ">Figure 9
<p>Structural characterization of ASPS−40, ASPS−60, and ASPS−80. (<b>a</b>) The λmax of Congo red + polysaccharides at various NaOH concentrations; (<b>b</b>) UV−vis spectrum in the presence of I<sub>2</sub>−KI.</p> Full article ">Figure 10
<p>Antioxidant test of ASPS−40, ASPS−60, and ASPS−80. (<b>a</b>) ABTS radical scavenging activities; (<b>b</b>) DPPH radical scavenging activities; (<b>c</b>) H<sub>2</sub>O<sub>2</sub> radical scavenging activities; (<b>d</b>) Fe<sup>2+</sup> chelating activities.</p> Full article ">
23 pages, 2588 KiB  
Article
Effect of Wild Strawberry Tree and Hawthorn Extracts Fortification on Functional, Physicochemical, Microbiological, and Sensory Properties of Yogurt
by Teresa Herrera, Maite Iriondo-DeHond, Ana Ramos Sanz, Ana Isabel Bautista and Eugenio Miguel
Foods 2023, 12(18), 3332; https://doi.org/10.3390/foods12183332 - 5 Sep 2023
Cited by 5 | Viewed by 2136
Abstract
The composition analyses and health-promoting properties (antioxidant capacity, antidiabetic, and antihypertensive properties) of wild fruit extracts and the effect of the incorporation of strawberry tree (STE) and hawthorn (HTE) extracts on the physicochemical, instrumental textural, microbiological, and sensory parameters of yogurts were evaluated. [...] Read more.
The composition analyses and health-promoting properties (antioxidant capacity, antidiabetic, and antihypertensive properties) of wild fruit extracts and the effect of the incorporation of strawberry tree (STE) and hawthorn (HTE) extracts on the physicochemical, instrumental textural, microbiological, and sensory parameters of yogurts were evaluated. The incorporation of wild fruit extracts in yogurt increased antioxidant and antidiabetic properties (inhibition of digestive α-amylase, α-glucosidase, and lipase enzymatic activities) compared to the control, without decreasing their sensory quality or acceptance by consumers. The hawthorn yogurt (YHTE) showed the highest total phenolic content (TPC) and antioxidant capacity (ABTS and ORAC methods). Yogurts containing wild fruit extracts and dietary fiber achieved high overall acceptance scores (6.16–7.04) and showed stable physicochemical, textural, and microbiological properties. Therefore, the use of wild fruit extracts and inulin-type fructans as ingredients in yogurt manufacture stands as a first step towards the development of non-added sugar dairy foods for sustainable health. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Instrumental firmness (N) and consistency (Ns) of a control yogurt (C), yogurt with strawberry tree extract (YSTE-8 and YSTE-12), and yogurt with hawthorn extract (YHE-8 and YHE-12). Different letters denote significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Spider-web diagram, which shows mean scores (<span class="html-italic">n</span> = 104) of a 1–10 scale sensorial analysis of yogurts: (<b>a</b>) Smell; (<b>b</b>) Taste; (<b>c</b>) Texture; (<b>d</b>) Overall acceptance. Different letters denote significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
13 pages, 2547 KiB  
Article
Analysis of the Flavonoidome Reveals the Different Health-Promoting Flavonoid Characteristics in Fruit
by Chi Zhang, Yao Zhao, Han Tao, Linying Li, Yuqing He, Xueying Zhang, Ying Zhu and Gaojie Hong
Antioxidants 2023, 12(9), 1665; https://doi.org/10.3390/antiox12091665 - 24 Aug 2023
Cited by 4 | Viewed by 2064
Abstract
Flavonoids are one of the important metabolites of plants, and many flavonoids have functions of antioxidant or antimicrobial, which can help plants resist environmental stress. On the other hand, flavonoids also have a health-promoting effect for humans, such as antioxidant and anti-aging, and [...] Read more.
Flavonoids are one of the important metabolites of plants, and many flavonoids have functions of antioxidant or antimicrobial, which can help plants resist environmental stress. On the other hand, flavonoids also have a health-promoting effect for humans, such as antioxidant and anti-aging, and some flavonoids can assist in disease treatment. Fruit is one of the main sources of plant food and flavonoids intake for humans. Understanding the flavonoidome of various fruits is helpful to choose fruit combinations according to different demands. In this study, we explored the composition and relative content of flavonoids in 22 fruits and analyzed some health-promoting flavonoids in fruits. In addition, we selected several fruits and measured their antioxidant capacity through experiments. Our study initially established a database of fruit flavonoidome, and can provide reference for nutrition research, fruit breeding and industrial development. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Principal component analysis (PCA) of fruit flavonoids. (<b>A</b>) Group of 22 fruits according to their ripening process. (<b>B</b>) PCA of the total flavonoids in 22 fruits.</p> Full article ">Figure 2
<p>Classification and statistics of flavonoids in fruit. (<b>A</b>) Classification of the total detected flavonoids. (<b>B</b>) Statistics and examples of flavonoids. (<b>C</b>) Structural formula of flavonoids in (<b>B</b>).</p> Full article ">Figure 3
<p>Distribution of flavonoid species in each fruit. (<b>A</b>) Cluster heatmap of the total flavonoids. (<b>B</b>) Cluster heatmap of the total flavonoids. (<b>C</b>) Cluster heatmap of the total flavonols. (<b>D</b>) Cluster heatmap of the total anthocyanins. (<b>E</b>) Cluster heatmap of the total isoflavones. (<b>F</b>) The total number of flavonoids in each fruit.</p> Full article ">Figure 4
<p>Some common health-promoting flavonoids in fruit.</p> Full article ">Figure 5
<p>The activity of the antioxidant enzymes in the different fruits. (<b>A</b>) Scavenging rate of the hydroxyl radical in the fruits. (<b>B</b>) CAT activity in the fruits. (<b>C</b>) SOD activity in the fruits. (<b>D</b>) POD activity in the fruits. Error bars represent the SE. Letters indicate significant difference assessed by one-way ANOVA, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">
19 pages, 4985 KiB  
Article
Salt Stress-Induced Modulation of Porphyrin Biosynthesis, Photoprotection, and Antioxidant Properties in Rice Plants (Oryza sativa)
by Anh Trung Nguyen, Lien Hong Tran and Sunyo Jung
Antioxidants 2023, 12(8), 1618; https://doi.org/10.3390/antiox12081618 - 15 Aug 2023
Cited by 2 | Viewed by 1553
Abstract
Salt stress disrupts cellular ion homeostasis and adversely impacts plant growth and productivity. We examined the regulatory mechanisms of porphyrin biosynthesis, photoprotection, and antioxidant properties in salt-stressed rice seedlings. In response to 150 mM NaCl, the rice seedlings exhibited dehydration, reduced relative water [...] Read more.
Salt stress disrupts cellular ion homeostasis and adversely impacts plant growth and productivity. We examined the regulatory mechanisms of porphyrin biosynthesis, photoprotection, and antioxidant properties in salt-stressed rice seedlings. In response to 150 mM NaCl, the rice seedlings exhibited dehydration, reduced relative water content, and increased levels of conductivity, malondialdehyde, and H2O2. The expression levels of the salt-stress-responsive genes NHX1, SOS1, and MYB drastically increased after NaCl treatment. The seedlings grown under NaCl stress displayed declines in Fv/Fm, ΦPSII, rETRmax, and photochemical quenching but increases in nonphotochemical quenching (NPQ) and the expression of genes involved in zeaxanthin formation, BCH, and VDE. Under salt stress conditions, levels of chlorophyll precursors significantly decreased compared to controls, matching the downregulation of CHLD, CHLH, CHLI, and PORB. By contrast, NaCl treatment led to increased heme content at 24 h of treatment and significant upregulations of FC2, HO1, and HO2 compared to controls. Salt-stressed seedlings also increased their expression of CATs (catalases) and APXs (ascorbate peroxidases) as well as the activities of superoxide dismutase, CAT, APX, and peroxidase. Our results indicate that chlorophyll and heme biosynthesis involve the protective strategies for salt stress alleviation through photoprotection by the scavenging of chlorophyll precursors and NPQ as well as activating antioxidant enzymes. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Oxidative metabolism in leaves of rice seedlings exposed to salt stress. (<b>A</b>) Phenotypes associated with NaCl-stress-related symptoms. (<b>B</b>) RWC of leaves. (<b>C</b>) Conductivity change. (<b>D</b>) MDA levels. (<b>E</b>) H<sub>2</sub>O<sub>2</sub>-DAB-staining in leaves. Brown spots indicate H<sub>2</sub>O<sub>2</sub> localization. The 14-day-old rice seedlings were transferred to half-strength Hoagland solution without or with 150 mM NaCl for 3 days. Control; half-strength Hoagland solution. 0, 24, 48, and 72; 0, 24, 48, and 72 h after NaCl treatment, respectively. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test. DW: dry weight.</p> Full article ">Figure 2
<p>Expression of stress-responsive genes induced by salt stress in leaves of rice seedlings. (<b>A</b>) Salt-stress-responsive genes. (<b>B</b>) Photosynthetic genes. (<b>C</b>) Genes involved in zeaxanthin formation. <span class="html-italic">Actin</span> was used as an internal control. The control sample was used as a calibrator, with its expression level set to 1. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">Figure 3
<p>Effects of salt stress on structure and function of photosystems and chlorophyll fluorescence quenching parameters in rice seedlings. (<b>A</b>) RLCs of the rETR. (<b>B</b>) Immunoblot analysis of the light-harvesting chlorophyll-binding proteins of PSII. Alpha-tubulin was used as a loading control. The relative expression levels of the LHCB proteins were quantified using ImageJ and normalized to Tubulin. (<b>C</b>) <span class="html-italic">F</span><sub>v</sub>/<span class="html-italic">F</span><sub>m</sub>. (<b>D</b>) Φ<sub>PSII</sub>. (<b>E</b>,<b>F</b>) Quenching parameters, qP and NPQ. Data are means ± SE of nine replicates obtained from three independent experiments (except six replicates from three independent experiments for immunoblot analysis). Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">Figure 4
<p>Effects of salt stress on metabolic intermediates of the common and chlorophyll branches in the porphyrin biosynthetic pathway in leaves of rice seedlings. (<b>A</b>) Chlorophyll. (<b>B</b>) ALA-synthesizing capacity. (<b>C</b>) Proto IX and Mg-porphyrin intermediates. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">Figure 5
<p>Salt-stress-induced changes in the expression of genes encoding enzymes of the chlorophyll biosynthetic pathway in leaves of rice seedlings. (<b>A</b>) Common branch. (<b>B</b>) Chlorophyll branch. <span class="html-italic">Actin</span> was used as an internal control. The control sample was used as a calibrator, with its expression level set to 1. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">Figure 6
<p>Effects of salt stress on the expression of genes encoding enzymes of the heme branch and heme content in leaves of rice seedlings. (<b>A</b>) Expression of genes in the heme branch. (<b>B</b>) Heme content. <span class="html-italic">Actin</span> was used as an internal control. The control sample was used as a calibrator, with its expression level set to 1. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">Figure 7
<p>Isozyme profiles of antioxidant enzymes in rice seedlings grown under salt stress (<b>A</b>) SOD. (<b>B</b>) CAT. (<b>C</b>) APX. (<b>D</b>) POD. The plants were subjected to the same treatments as in <a href="#antioxidants-12-01618-f001" class="html-fig">Figure 1</a>. The numbers show each isozyme of antioxidant enzymes in order of bands from the top.</p> Full article ">Figure 8
<p>Effects of salt stress on the expression of genes encoding the H<sub>2</sub>O<sub>2</sub>-scavenging enzymes and redox state of ascorbate. (<b>A</b>) <span class="html-italic">CATs</span>. (<b>B</b>) <span class="html-italic">APXs</span>. (<b>C</b>) Redox state of ascorbate: estimated as Asc × 100/Asc<sub>t</sub>, where Asc<sub>t</sub> = DHA + Asc. <span class="html-italic">Actin</span> was used as an internal control. The control sample was used as a calibrator, with its expression level set to 1. Data are means ± SE of nine replicates obtained from three independent experiments. Mean values followed by different lowercase letters are significantly different at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test.</p> Full article ">
18 pages, 2313 KiB  
Article
On the Role of ROS and Glutathione in the Mode of Action Underlying Nrf2 Activation by the Hydroxyanthraquinone Purpurin
by Qiuhui Ren, Wouter Bakker, Sebastiaan Wesseling, Hans Bouwmeester and Ivonne M. C. M. Rietjens
Antioxidants 2023, 12(8), 1544; https://doi.org/10.3390/antiox12081544 - 2 Aug 2023
Cited by 3 | Viewed by 1530
Abstract
Purpurin is a major anthraquinone present in the roots of Rubia cordifolia (madder). Purpurin is known to activate Nrf2 (Nuclear transcription factor erythroid 2-related factor 2) EpRE (electrophile responsive element) mediated gene expression as a potential beneficial effect. This study aimed to elucidate [...] Read more.
Purpurin is a major anthraquinone present in the roots of Rubia cordifolia (madder). Purpurin is known to activate Nrf2 (Nuclear transcription factor erythroid 2-related factor 2) EpRE (electrophile responsive element) mediated gene expression as a potential beneficial effect. This study aimed to elucidate the balance between the electrophilicity or pro-oxidant activity of purpurin underlying the Nrf2 induction. For this, Nrf2 activation with modified intracellular glutathione (GSH) levels was measured in an Nrf2 CALUX reporter gene assay. In addition, both cell-free and intracellular ROS formation of purpurin with modified (intracellular) GSH levels at different pH were quantified using the DCF-DA assay. GSH adduct formation was evaluated by UPLC and LC-TOF-MS analysis. GSH and GSSG levels following purpurin incubations were quantified by LC-MS/MS. We show that Nrf2 induction by purpurin was significantly increased in cells with buthionine sulfoximine depleted GSH levels, while Nrf2 induction was decreased upon incubation of the cells with N-acetylcysteine being a precursor of GSH. In cell-free incubations, ROS formation increased with increasing pH pointing at a role for the deprotonated form of purpurin. Upon incubations of purpurin with GSH at physiological pH, GSH adduct formation appeared negligible (<1.5% of the added purpurin). The addition of GSH resulted in conversion of GSH to GSSG and significantly reduced the ROS formation. Together these results demonstrate that Nrf2 induction by purpurin originates from intracellular ROS formation and not from its electrophilicity, which becomes especially relevant when intracellular GSH levels can no longer scavenge the ROS. The present study demonstrated that the efficiency of intracellular Nrf2 activation by purpurin and related anthraquinones will depend on (i) their pKa and level of deprotonation at the intracellular pH, (ii) the oxidation potential of their deprotonated form and (iii) the intracellular GSH levels. Thus, the Nrf2 induction by purpurin depends on its pro-oxidant activity and not on its electrophilicity. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Potential modes of action underlying purpurin-induced induction of Nrf2-mediated gene expression via either electrophilicity or redox cycling and ROS formation.</p> Full article ">Figure 2
<p>Concentration-dependent effect of purpurin on (<b>A</b>) Nrf2-mediated gene expression in Nrf2 CALUX cells treated with purpurin at different concentrations for 24 h and (<b>B</b>) intracellular ROS formation in Nrf2 CALUX cells treated with purpurin at different concentrations for 4 h. The ROS formation in the solvent control was set as 1 and indicated by the dashed line. Data are shown as mean fold induction or fold increase compared to solvent control ± SEM of 3 replicates. (***, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">Figure 3
<p>(<b>A</b>) Nrf2 activation and (<b>B</b>) viability of Nrf2 CALUX cells exposed to 1% DMSO (solvent control) and different concentrations of purpurin in the absence (black bars) or presence of 30 µM of BSO (Red bars), or with 5 mM of NAC (blue bars). The Luciferase induction was expressed as induction factor (IF) compared to the solvent control. The data are presented as mean ± SEM of three independent replicates and Asterisk indicates a response significantly different from treatment of solvent control (1% DMSO) or treatment of purpurin at the respective concentration (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">Figure 4
<p>pKa values of purpurin predicted by MolGpka obtained through the web server. (<a href="https://xundrug.cn/molgpka" target="_blank">https://xundrug.cn/molgpka</a>, accessed on 17 August 2022).</p> Full article ">Figure 5
<p>Representative part of the HPLC-UV chromatograms of incubations of purpurin without (<b>A</b>) with (<b>B</b>) GSH at pH 8. The peaks labeled in a, b, c exist in both incubation samples without or with GSH at pH8 and originate from purpurin, representing purpurin itself (peak labelled a) and impurities present in the commercial purpurin preparation already present at the start of the incubation (peaks b and c). These peaks were not identified to a further extent since the aim of the experiment was to show the (limited) formation of the GS-purpurin adduct. The metabolite eluting at 4.25 min was shown by LC-TOF-MS analysis to have an <span class="html-italic">m</span>/<span class="html-italic">z</span> of 560.1 representing a glutathione adduct of purpurin.</p> Full article ">Figure 6
<p>Time-dependent changes in GSH levels, GSSG levels and recovery in incubations of purpurin (200 µM) without or with GSH (200 µM) at pH 5 (<b>A</b>), 6 (<b>B</b>), 7 (<b>C</b>) and 8 (<b>D</b>). The data presented as mean ± SEM of at least three independent replicates. GSH_SC: GSH solvent control, GSSG_SC: GSSG solvent control. * in black indicates a response of GSH depletion significantly different from incubations in the absence of purpurin; and # in red indicates a response of GSSG formation significantly different from incubations in the absence of purpurin (* and #, <span class="html-italic">p</span> &lt; 0.05; ** and ##, <span class="html-italic">p</span> &lt; 0.01; *** and ###, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">Figure 7
<p>ROS formation in cell-free incubations of purpurin (200 µM) at different pH values. Data are shown as mean fold increase compared to control ± SEM of 3 replicates and Asterisk indicates a response significantly different from treatment of solvent control (1% DMSO) (**, <span class="html-italic">p</span> &lt; 0.01; ***, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">Figure 8
<p>Reactive oxygen species (ROS) measured in a cell-free system with 200 µM purpurin incubated in the absence (Red bars) and presence of 5 mM GSH (Orange bars) and 10 mM GSH (Yellow bars). Data are shown as mean fold increase relative to control ± SEM of 3 replicates and Asterisk indicates a response significantly different from treatment of solvent control (1% DMSO) (black bars) or treatment of 200 µM purpurin alone at different pH (***, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">Figure 9
<p>200 µM purpurin induced formation of reactive oxygen species (ROS) in a cell-free system in the presence of GSH at different concentrations (0.1, 0.5, 1, 5 and 10 mM) at pH 5, 6, 7 and 8. Data are shown as mean fold increase relative to control ± SEM of 3 replicates and Asterisk indicates a response significantly different from treatment of solvent control (1% DMSO) or treatment of purpurin alone (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01;***, <span class="html-italic">p</span> &lt; 0.001; one-way ANOVA analysis with post-hoc Tukey test).</p> Full article ">
16 pages, 800 KiB  
Article
Stability of Phenols, Antioxidant Capacity and Grain Yield of Six Rice Genotypes
by Juthathip Kunnam, Wanwipa Pinta, Ruttanachira Ruttanaprasert, Darika Bunphan, Thanasin Thabthimtho and Chorkaew Aninbon
Plants 2023, 12(15), 2787; https://doi.org/10.3390/plants12152787 - 27 Jul 2023
Cited by 5 | Viewed by 1212
Abstract
The environment is the main factor affecting variations in phytochemicals and antioxidant activity in rice. The objective of this study was to evaluate the stability of grain yield, phytochemicals and antioxidant capacity of six rice genotypes. Six rice genotypes were evaluated in a [...] Read more.
The environment is the main factor affecting variations in phytochemicals and antioxidant activity in rice. The objective of this study was to evaluate the stability of grain yield, phytochemicals and antioxidant capacity of six rice genotypes. Six rice genotypes were evaluated in a randomized complete block design with three replicates at three locations in Trat, Bangkok and Sakon Nakhon provinces in July–October 2019. Data on grain yield, yield components, total phenolic content, ferulic acid and antioxidant capacity were recorded. Grain yield was highest for crops grown in Bangkok, whereas antioxidant activity was highest for crops grown in Bangkok and Sakon Nakhon. Hom Nang Nual 1 and Mali Nil Boran had the highest grain yield. Riceberry had the highest grain yield in Trat; it also had high levels of total phenolic compounds, ferulic acid and antioxidant activity. Mali Nil Boran, Mali Nil Surin and Riceberry had the most stable total phenolic content, ferulic acid and antioxidant activity, respectively. Information on the levels and variability of phytochemicals in rice enables the selection of genotypes with high and stabile phytochemicals for production and rice breeding. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Maximum temperature (Tmax; °C), minimum temperature (Tmin; °C), relative humidity (%) and rainfall (mm) in Bangkok (<b>a</b>,<b>d</b>), Trat (<b>b</b>,<b>e</b>) and Sakon Nakhon (<b>c</b>,<b>f</b>) provinces during the growing season.</p> Full article ">
15 pages, 2652 KiB  
Article
Effect of the Addition of Freeze-Dried Grape Pomace on Fresh Tagliatelle Gluten Network and Relationship to Sensory and Chemical Quality
by Barbara la Gatta, Mariacinzia Rutigliano, Maria Teresa Liberatore, Flavia Dilucia, Maurizio Palmitessa, Aldo Di Luccia and Carmela Lamacchia
Foods 2023, 12(14), 2699; https://doi.org/10.3390/foods12142699 - 13 Jul 2023
Cited by 3 | Viewed by 1681
Abstract
The incorporation of 5 and 10% freeze-dried grape pomace powder (GPP) in fresh tagliatelle pasta preparation was evaluated for its effect on chemical composition, gluten protein structure, and sensory properties. The addition of the freeze-dried GPP led to a significant increase (p [...] Read more.
The incorporation of 5 and 10% freeze-dried grape pomace powder (GPP) in fresh tagliatelle pasta preparation was evaluated for its effect on chemical composition, gluten protein structure, and sensory properties. The addition of the freeze-dried GPP led to a significant increase (p < 0.05) in polyphenol content in the raw and cooked fortified pasta samples with respect to 100% semolina pasta, although the phenolic content decreased after the cooking process. The opposite phenomenon was observed with the antioxidant activity, which increased significantly (p < 0.05) when switching from raw to cooked pasta samples fortified with GPP. The formation of a proper gluten structure was found in the fortified raw pasta, even if a change in the protein arrangement was shown in the fortified cooked samples, confirmed by a significant reduction (p < 0.05) in both the unextractable polymeric protein percentage (% UPP) and disulfide bond (S-S) formation. These results suggest a possible interaction between the protein sulfhydryl groups (-Cys) and polyphenols of grape pomace during cooking through non-disulfide covalent bonds, which was confirmed by the significant (p < 0.05) decrease in the -SH groups when comparing 100% semolina pasta with fortified pasta sample. Finally, a sensory analysis showed that the highest significant score (p < 0.05) was achieved by the 5% GP-fresh pasta sample. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Fresh pasta samples. (<b>A</b>) 100% Durum wheat semolina fresh pasta; (<b>B</b>) 5% Grape pomace powder fresh pasta; (<b>C</b>) 10% Grape pomace powder fresh pasta.</p> Full article ">Figure 2
<p>SE-HPLC profiles of SDS-extractable proteins of samples. (<b>A</b>): semolina flour; (<b>B</b>): freeze-dried GPP; (<b>C</b>): raw fresh pasta samples; (<b>D</b>): cooked fresh pasta samples.</p> Full article ">Figure 3
<p>Unextractable polymeric proteins (%UPP) in raw and cooked pasta samples.</p> Full article ">Figure 4
<p>SDS-PAGE of fresh pasta samples. (<b>A</b>): Raw samples under reducing conditions; (<b>B</b>): Cooked samples under reducing conditions; (<b>C</b>): Raw samples under not reducing conditions; (<b>D</b>): Cooked samples under not reducing conditions. M: marker; SEM: semolina flour, 1: 100% Semolina fresh pasta; 2: 5% GP-fresh pasta; 3: 10% GP-fresh pasta.</p> Full article ">Figure 5
<p>Spider plot with mean values for technical attributes evaluated for fresh pasta sample.</p> Full article ">
17 pages, 1469 KiB  
Review
Mechanisms of Action of Fruit and Vegetable Phytochemicals in Colorectal Cancer Prevention
by Teresita Alzate-Yepes, Lorena Pérez-Palacio, Estefanía Martínez and Marlon Osorio
Molecules 2023, 28(11), 4322; https://doi.org/10.3390/molecules28114322 - 24 May 2023
Cited by 17 | Viewed by 4100
Abstract
Colorectal cancer (CRC) is the third most common cancer worldwide and its incidence is expected to increase by almost 80% by 2030. CRC apparition is related to poor diet, mainly due to low consumption of phytochemicals present in fruits and vegetables. Hence, this [...] Read more.
Colorectal cancer (CRC) is the third most common cancer worldwide and its incidence is expected to increase by almost 80% by 2030. CRC apparition is related to poor diet, mainly due to low consumption of phytochemicals present in fruits and vegetables. Hence, this paper reviews the most promising phytochemicals in the literature, presenting scientific evidence regarding potential CRC chemopreventive effects. Moreover, this paper reveals the structure and action of CRC mechanisms that these phytochemicals are involved in. The review reveals that vegetables rich in phytochemicals such as carrots and green leafy vegetables, as well as some fruits such as pineapple, citrus fruits, papaya, mango, and Cape gooseberry, that have antioxidant, anti-inflammatory, and chemopreventive properties can promote a healthy colonic environment. Fruits and vegetables in the daily diet promote antitumor mechanisms by regulating cell signaling and/or proliferation pathways. Hence, daily consumption of these plant products is recommended to reduce the risk of CRC. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Basic structures of phenolic acids, polyphenols and flavonoids, and their derivatives implicated as chemopreventive compounds.</p> Full article ">Figure 2
<p>Mechanisms of anticancer action of phytochemicals for colorectal cancer chemoprevention. TNFα: tumor necrosis factor, TNF-TNFR: tumor necrosis factor-tumor necrosis factor receptor complex, TRADD: tumor necrosis factor receptor type 1, TRAF2: TNF receptor-associated factor 2, FADD: FAS-associated death domain protein, RIP: ribosome-inactivating protein, IkBa: inhibitor of nuclear factor kappa-B kinase subunit alpha, p65 and p50: Transcription factors 65 and 50, respectively, ROS: reactive oxygen species MAPK: mitogen-activated protein kinase, RAS: guanosine-nucleotide-binding protein, MEKs: mitogen-activated protein kinase kinase, RAF: serine/threonine-specific protein kinases, p38: mitogen-activated protein kinases, JNK: c-Jun N-terminal kinase, ERK: extracellular signal-regulated kinase, P21: cyclin-dependent kinase inhibitor, CDK2: cell division protein kinase 2.</p> Full article ">Figure 3
<p>Chemical structures of non-polyphenolic compounds.</p> Full article ">
19 pages, 4634 KiB  
Article
Modification of the Structural and Functional Characteristics of Mung Bean Globin Polyphenol Complexes: Exploration under Heat Treatment Conditions
by Yantao Ma, Shu Zhang, Yuchao Feng, Haoyu Wang, Yuhang Liu and Changyuan Wang
Foods 2023, 12(11), 2091; https://doi.org/10.3390/foods12112091 - 23 May 2023
Cited by 7 | Viewed by 1835
Abstract
During the storage and processing of mung beans, proteins and polyphenols are highly susceptible to interactions with each other. Using globulin extracted from mung beans as the raw material, the study combined it with ferulic acid (FA; phenolic acid) and vitexin (flavonoid). Physical [...] Read more.
During the storage and processing of mung beans, proteins and polyphenols are highly susceptible to interactions with each other. Using globulin extracted from mung beans as the raw material, the study combined it with ferulic acid (FA; phenolic acid) and vitexin (flavonoid). Physical and chemical indicators were combined with spectroscopy and kinetic methods, relying on SPSS and peak fit data to statistically analyze the conformational and antioxidant activity changes of mung bean globulin and two polyphenol complexes before and after heat treatment and clarify the differences and the interaction mechanism between globulin and the two polyphenols. The results showed that, with the increase in polyphenol concentration, the antioxidant activity of the two compounds increased significantly. In addition, the antioxidant activity of the mung bean globulin–FA complex was stronger. However, after heat treatment, the antioxidant activity of the two compounds decreased significantly. The interaction mechanism of the mung bean globulin–FA/vitexin complex was static quenching, and heat treatment accelerated the occurrence of the quenching phenomenon. Mung bean globulin and two polyphenols were combined through a hydrophobic interaction. However, after heat treatment, the binding mode with vitexin changed to an electrostatic interaction. The infrared characteristic absorption peaks of the two compounds shifted to different degrees, and new peaks appeared in the areas of 827 cm−1, 1332 cm−1, and 812 cm−1. Following the interaction between mung bean globulin and FA/vitexin, the particle size decreased, the absolute value of zeta potential increased, and the surface hydrophobicity decreased. After heat treatment, the particle size and zeta potential of the two composites decreased significantly, and the surface hydrophobicity and stability increased significantly. The antioxidation and thermal stability of the mung bean globulin–FA were better than those of the mung bean globulin–vitexin complex. This study aimed to provide a theoretical reference for the protein–polyphenol interaction mechanism and a theoretical basis for the research and development of mung bean functional foods. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>(<b>A</b>) The binding rate and mass ratio of mung bean globulin to vitexin (20, 40, 60, 80, and 100 μmol/g pro) and ferulic acid (FA) (40, 60, 80, 100, and 120 μmol/g pro) at different ratios; (<b>B</b>) Binding rate and mass ratio of mung bean globulin and FA at of 25, 70, 85, and 100 °C; (<b>C</b>) Binding rate and mass ratio of mung bean globulin and vitexin at 25, 70, 85, and 100 °C. Note: Different letters indicate differences. There were significant differences between the groups (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>(<b>A</b>) solubility of the complexes at different ratios; (<b>B</b>) solubility of the complexes at different temperatures. The upper case letters in Figure (<b>A</b>) show the significance between the pv groups and the lower case letters show the difference between the PFA groups; the lower case letters in Figure (<b>B</b>) show the significance between the pv groups and the upper case letters show the difference between the PFA groups.</p> Full article ">Figure 3
<p>(<b>A</b>) ABTS+ and DPPH of the mung bean globulin–ferulic acid complex in different proportions; (<b>B</b>) ABTS+ and DPPH of the mung bean globulin–vitexin complex at different proportions. Different letters within a column indicate significant differences for each parameter (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Two-dimensional fluorescence spectrum: (<b>A</b>) Mung bean globulin–ferulic acid complex in different proportions; (<b>B</b>) Mung bean globulin–vitexin complex in different proportions; (<b>C</b>) Mung bean globulin–ferulic acid complex at different temperatures; (<b>D</b>) Mung bean globulin–vitexin complex at different temperatures.</p> Full article ">Figure 5
<p>Three-dimensional fluorescence spectrum: (<b>A</b>) Mung bean globulin–ferulic acid complex in different proportions; (<b>B</b>) Mung bean globulin–vitexin complex in different proportions; (<b>C</b>) Effect of temperature on the mung bean globulin–polyphenol complex; 25–100 represent mung bean globulin–ferulic acid complexes, 25′–100′ represent mung bean globulin–vitexin complexes.</p> Full article ">Figure 6
<p>Stern–Volmer equation and double logarithmic diagram of the interaction between mung bean globulin and polyphenols at 298 K (25 °C), 343 K (70 °C), 353 K (80 °C), 363 K (90 °C), and 373 K (100 °C): (<b>A</b>) PFA; (<b>B</b>) PV.</p> Full article ">Figure 7
<p>Fourier infrared spectroscopy. (<b>A</b>,<b>B</b>) Mung bean globulin–ferulic acid complex and mung bean globulin–vitexin complex at different ratios; (<b>C</b>,<b>D</b>) Mung bean globulin–Ferulic acid complex and mung bean globulin–vitexin complex at different temperatures.</p> Full article ">Figure 8
<p>SDS–PAGE electrophoretogram: (<b>A</b>) Mung bean globulin–ferulic acid complex in different proportions; (<b>B</b>) Mung bean globulin–vitexin complex in different proportions; (<b>C</b>) Mung bean globulin–ferulic acid complex at different temperatures; (<b>D</b>) Mung bean globulin–vitexin complex at different temperatures.</p> Full article ">Figure 9
<p>Particle size and ζ potential plots: (<b>A</b>) Mung bean globulin–ferulic acid complex in different proportions; (<b>B</b>) Mung bean globulin–vitexin complex in different proportions; (<b>C</b>) Mung bean globulin–ferulic acid complex at different temperatures; (<b>D</b>) Mung bean globulin–vitexin complex at different temperatures. Different letters within a picture indicate significant differences for each parameter (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 10
<p>(<b>A</b>) Surface hydrophobicity of mung bean globulin after reaction with different concentrations of polyphenols; (<b>B</b>) Effect of heat treatment on surface hydrophobicity of the mung bean globulin–polyphenol complex. Note: Different letters indicate differences, and there were significant differences between the groups (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
19 pages, 6399 KiB  
Article
Comprehensive Assessment of the Antioxidant and Anticancer Potential of Selected Ethnobotanical Plants
by Monoj Kumar Das, Neelu Singh and Paulraj Rajamani
Oxygen 2023, 3(2), 203-221; https://doi.org/10.3390/oxygen3020015 - 22 May 2023
Viewed by 1921
Abstract
Globally, among different types of cancers, breast cancer is identified as the chief cause of mortality among females, and it is a challenge to find new effective treatment strategies with minimal side effects and increased efficacy. Plants are an integral part of the [...] Read more.
Globally, among different types of cancers, breast cancer is identified as the chief cause of mortality among females, and it is a challenge to find new effective treatment strategies with minimal side effects and increased efficacy. Plants are an integral part of the traditional indigenous healthcare system and are becoming the concrete source of new drug discovery. Thus, there is a need to obtain a scientific basis for applying traditionally used plants in cancer treatments that may harbour novel phytochemicals. Therefore, this study aims to investigate the antioxidant and anticancer potential of selected plants of ethnobotanical importance. Five plants of ethnobotanical importance were selected and screened to determine their antioxidant potential through various in vitro free radical scavenging assays (such as DPPH, ABTS, hydroxyl, and superoxide radical scavenging), ferric chelation, and total antioxidant potential, and the total phenolic and flavonoid content was estimated for the selected plants. In contrast, the anticancer potential of crude plant extracts was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) against different breast cancer (MCF-7, MDA-MB-231, and MDA-MB-435S) and hepatic cancer cell lines (HepG2), and human PBMCs (peripheral blood mononuclear cells) were used for toxicity studies. The MTT results showed that among all of the crude plant extracts (CAN = Etlingera linguiformis, SES = Sesbania grandiflora, LEX = Smilax ovalifolia, DES = Desmodium triflorum, and CA = Chenopodium album), it was CAN and LEX that showed the best cytotoxic potential on exposed breast cancer cell lines in contrast to SES, DES, and CA. In addition, at the selected dosages that were exposed to breast cancer cells, none of the extracts from any of the five plants showed any cytotoxicity against human PBMCs. Thus, the crude extracts can be explored further for chemopreventive and anticancer activity on murine models to understand their underlying mechanism for effective cancer management. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Antioxidative and free radical scavenging activity of CAN, LEX, SES, DES, and CA for (<bold>a</bold>) ABTS, and (<bold>b</bold>) DPPH were evaluated using in vitro cell-free chemical-based reactions. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values were significant (<italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05) when compared to the control. BHA was taken as a positive control for both ABTS and DPPH radical activity.</p> Full article ">Figure 1 Cont.
<p>Antioxidative and free radical scavenging activity of CAN, LEX, SES, DES, and CA for (<bold>a</bold>) ABTS, and (<bold>b</bold>) DPPH were evaluated using in vitro cell-free chemical-based reactions. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values were significant (<italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05) when compared to the control. BHA was taken as a positive control for both ABTS and DPPH radical activity.</p> Full article ">Figure 2
<p>The antioxidative and free radical scavenging activity of CAN, LEX, SES, DES, and CA against (<bold>a</bold>) superoxide and (<bold>b</bold>) hydroxyl radicals were evaluated using in vitro cell-free chemical-based reactions. Values were expressed as mean ± SEM; <italic>n</italic> = 3; values are significant <sup>a</sup> <italic>p</italic> ≤ 0.001, <sup>b</sup> <italic>p</italic> ≤ 0.01, and <sup>c</sup> <italic>p</italic> ≤ 0.05 when compared to the control. Ascorbic acid and gallic acid were used as a positive control for both superoxide and hydroxyl radical activity.</p> Full article ">Figure 3
<p>The Fe<sup>2+</sup> chelation inhibitory activities of CAN, LEX, SES, DES, and CA were evaluated using in vitro cell-free chemical-based reactions. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values were significant <italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05 when compared to the control. EDTA was used as a positive control.</p> Full article ">Figure 4
<p>Fe<sup>3+</sup> reducing ability of CAN, LEX, SES, DES, and CA: The reductive ability of CAN, LEX, SES, DES, CA, and ascorbic acid was evaluated using in vitro cell-free chemical-based reactions. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values are significant when <italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05 compared to the control. Ascorbic acid was taken as the positive control.</p> Full article ">Figure 5
<p>The effect of cell viability of CAN, LEX, SES, DES, and CA in HepG2 cells and breast cancer cell lines (MCF-7, MDA-MB-231, and MDA-MB-435S) at 48 h, measured using an MTT-based method. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values are significant when <italic><sup>a</sup> p</italic> ≤ 0.0001, <italic><sup>b</sup> p</italic> ≤ 0.001, <italic><sup>c</sup> p</italic> ≤ 0.01 and <italic><sup>d</sup> p</italic> ≤ 0.05 compared to the control.</p> Full article ">Figure 6
<p>The effect on cell viability of CAN, LEX, SES, DES, and CA in PBMC cells at 12, 18, and 36 h, measured using an MTT-based method. Values are expressed as mean ± SEM; <italic>n</italic> = 3; values were significant when <italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05 compared to the control.</p> Full article ">Figure 7
<p>The effect on cell viability of CAN, LEX, DES, SES, and CA in HepG2 cells, measured at 48 h using an MTT-based method. Values are expressed as the mean ± SEM; <italic>n</italic> = 3; All of the values were compared with both negative (0.1% DMSO) and positive (0.2% H<sub>2</sub>O<sub>2</sub>) controls. Values are significant (<italic><sup>a</sup> p</italic> ≤ 0.001, <italic><sup>b</sup> p</italic> ≤ 0.01, and <italic><sup>c</sup> p</italic> ≤ 0.05) for the negative control.</p> Full article ">Figure 8
<p>The effect of reactive oxygen species (ROS) on CAN, LEX, DES, SES, and CA in HepG2 cells at 48 h, measured using a fluorometric-based method. The relative fluorescence unit is calculated as a fold change relative to the negative control and is expressed as the mean ± SD, <italic>n</italic> = 3. All of the values were compared to both negative (0.1% DMSO) and positive (0.2% H<sub>2</sub>O<sub>2</sub>) controls. Values were significant (<italic><sup>a</sup> p</italic> ≤ 0.001) for the negative control and (<italic><sup>x</sup> p</italic> ≤ 0.001) for the positive control.</p> Full article ">
14 pages, 4857 KiB  
Article
Toxicity Effects of Polystyrene Nanoplastics with Different Sizes on Freshwater Microalgae Chlorella vulgaris
by Qingqing Xiang, Ying Zhou and Chengxia Tan
Molecules 2023, 28(9), 3958; https://doi.org/10.3390/molecules28093958 - 8 May 2023
Cited by 10 | Viewed by 2659
Abstract
The ubiquitous nature of plastics, particularly nanoplastics, raises concern about their potential effects on primary producer microalgae. Currently, the impacts and potential mechanisms of nanoplastics on microalgae are not fully understood. In this study, the effects of two plain commercial polystyrene nanoplastics (PS-NPs) [...] Read more.
The ubiquitous nature of plastics, particularly nanoplastics, raises concern about their potential effects on primary producer microalgae. Currently, the impacts and potential mechanisms of nanoplastics on microalgae are not fully understood. In this study, the effects of two plain commercial polystyrene nanoplastics (PS-NPs) with different sizes (50 nm and 70 nm) on C. vulgaris were assessed in a concentration range of 0–50 mg/L during 72 h exposure periods. Results revealed that both PS-NPs have dose-dependent toxicity effects on C. vulgaris, as confirmed by the decrease of growth rates, chlorophyll a and esterase activities, and the increase of ROS, MDA, and membrane damage. The membrane damage was caused by the agglomeration of PS-NPs on microalgae and may be the key reason for the toxicity. Compared with 70 nm PS-NPs (72 h EC50 >50 mg/L), 50 nm PS-NPs posed greater adverse effects on algae, with an EC50–72h of 19.89 mg/L. FTIR results also proved the stronger variation of macromolecules in the 50 nm PS-NPs treatment group. This phenomenon might be related to the properties of PS-NPs in exposure medium. The lower absolute zeta potential value of 50 nm PS-NPs induced the stronger interaction between PS-NPs and algae as compared to 70 nm PS-NPs, leading to severe membrane damage and the loss of esterase activity as well as settlement. These findings emphasized the importance of considering the impacts of commercial PS-NPs properties in toxicity evaluation. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Full article ">Figure 1
<p>Intensity-based size distributions by DLS analysis of 50 nm (<b>a</b>) and 70 nm (<b>b</b>) PS-NPs (10 mg/L).</p> Full article ">Figure 2
<p>Effects of 50 nm (<b>a</b>,<b>c</b>) and 70 nm (<b>b</b>,<b>d</b>) PS-NPs on <span class="html-italic">C. vulgaris</span> growth during 72 h exposure durations. Significant differences between the control and treatments are marked with an asterisk (*: <span class="html-italic">p</span>&lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01).</p> Full article ">Figure 3
<p>Effects of 50 nm (<b>a</b>) and 70 nm (<b>b</b>) PS-NPs on chlorophyll a during 72 h exposure durations. Significant differences between the control and treatments are marked with an asterisk (*: <span class="html-italic">p</span> &lt; 0.05, **: <span class="html-italic">p</span> &lt; 0.01).</p> Full article ">Figure 4
<p>SEM images of <span class="html-italic">C. vulgaris</span> after 72 h exposure to 50 nm (<b>above</b>) and 70 nm (<b>below</b>) PS-NPs.</p> Full article ">Figure 5
<p>Variations of ROS relative content (<b>a</b>), CAT relative content (<b>b</b>), and MDA content (<b>c</b>) after treating with different concentrations of PS-NPs (* and ** denote significant (<span class="html-italic">p</span> &lt; 0.05) and very significant (<span class="html-italic">p</span> &lt; 0.01) differences from the control, respectively).</p> Full article ">Figure 6
<p>Alternation of esterase activity(FL1 fluorescence) of <span class="html-italic">C. vulgaris</span> after 72 h exposure to PS-NPs by flow cytometry: (<b>a</b>) control, (<b>b</b>) 10 mg/L, (<b>c</b>) 20 mg/L, (<b>d</b>) 50 mg/L.</p> Full article ">Figure 7
<p>FTIR spectrums of <span class="html-italic">C. vulgaris</span> after treating with 20 mg/L PS-NPs (black line: control, red line: 50 nm PS-NPs, blue line: 70 nm PS-NPs).</p> Full article ">Figure 8
<p>Microscope images of <span class="html-italic">C. vulgaris</span> after 72 h exposure to 50 nm (<b>lower</b>) and 70 nm (<b>upper</b>) PS-NPs (40× magnification). The fluorescent images (black part) were obtained under the excitation mode of green light source. Arrows show the aggregation of <span class="html-italic">C. vulgaris</span> and appear orange fluorescent.</p> Full article ">
18 pages, 2972 KiB  
Article
Pomegranate Fruit Cracking during Maturation: From Waste to Valuable Fruits
by Caterina Fraschetti, Enkelejda Goci, Alexandru Nicolescu, Francesco Cairone, Simone Carradori, Antonello Filippi, Vincenzo Palmieri, Andrei Mocan and Stefania Cesa
Foods 2023, 12(9), 1908; https://doi.org/10.3390/foods12091908 - 6 May 2023
Cited by 4 | Viewed by 3297
Abstract
The pomegranate is an emerging functional food which is nowadays becoming more and more commercially attractive. Each part of this fruit (peels, arils and seeds) has a specific phytocomplex, rich in anti-oxidant and anti-radical compounds. Among these, punicalagin and ellagic acid continue to [...] Read more.
The pomegranate is an emerging functional food which is nowadays becoming more and more commercially attractive. Each part of this fruit (peels, arils and seeds) has a specific phytocomplex, rich in anti-oxidant and anti-radical compounds. Among these, punicalagin and ellagic acid continue to be widely studied for their numerous beneficial effects on human health (anti-inflammatory effects, anti-diabetes activity, cardio-protection, cancer prevention). Despite their exceptionally valuable composition and high adaptability to different climatic conditions, pomegranate fruits are highly susceptible to splitting during different stages of ripening, so much so that an estimated 65% of the production may be lost. A “zero-kilometer” approach should therefore be adopted to utilize such a valuable product otherwise destined to be downgraded or even incinerated, with a very high environmental impact. The aim of this work is to highlight and compare the compositional differences between whole and split pomegranates belonging to the cultivar Dente di Cavallo, grown in Apulia (Italy), to assess a valuable role for this split fruit usually considered as waste. The arils and peels are subjected to extraction procedures and the extracts analyzed by CIEL*a*b*, HPLC-DAD and HS-SPME/GC-MS. Moreover, an assessment of the inhibitory activity against α-glucosidase, acetylcholinesterase and tyrosinase enzymes has also been applied. The data show a better chemical profile in split fruits (namely 60% more anthocyanin content than intact fruit) with very interesting results in terms of α-glucosidase inhibition. The juices obtained by squeezing are also compared to commercial juices (“Salus Melagrana” and “La Marianna”) processed from the same cultivar and subjected to the same protocol analysis. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Flow chart.</p> Full article ">Figure 2
<p>Palette of analyzed samples (<b>A</b>); reflectance curves related to W and J (intact and split) samples (<b>B</b>); reflectance curves related to obtained hydroalcoholic extracts (<b>C</b>,<b>D</b>).</p> Full article ">Figure 3
<p>Shelf-life study on Salus Melagrana (J<sub>S</sub>) (<b>A</b>) and La Marianna (J<sub>L</sub>) juices (<b>B</b>).</p> Full article ">Figure 4
<p>Example of HA<sub>W split</sub> chromatograms at 280 nm (1. Gallic acid) and at 360 nm (2. α-punicalagin, 3. β-punicalagin, 4. ellagic acid), and HA<sub>WA split</sub> chromatograms at 520 nm for the identification of anthocyanins.</p> Full article ">Figure 5
<p>Effect of harvesting date on pomegranate phytocomplex.</p> Full article ">Figure 6
<p>Shelf life of “Salus Melagrana” and “La Marianna” juices.</p> Full article ">Figure 7
<p>Comparison in the distribution of the prevailing classes of compounds in split and intact HA<sub>J</sub> and HA<sub>W</sub>.</p> Full article ">Figure 8
<p>Distribution of the prevailing classes of compounds in HA<sub>P split</sub>.</p> Full article ">Figure 9
<p>Logarithmic inhibitory curves for the pomegranate samples, as well as for acarbose as positive control.</p> Full article ">
21 pages, 6834 KiB  
Article
The Hydrophobic Extract of Sorghum bicolor (L. Moench) Enriched in Apigenin-Protected Rats against Aflatoxin B1-Associated Hepatorenal Derangement
by Solomon E. Owumi, Blessing Ajakaiye, Adenike O. Akinwunmi, Sarah O. Nwozo and Adegboyega K. Oyelere
Molecules 2023, 28(7), 3013; https://doi.org/10.3390/molecules28073013 - 28 Mar 2023
Cited by 3 | Viewed by 1947
Abstract
Aflatoxin B1 (AFB1) is a recalcitrant metabolite produced by fungi species, and due to its intoxications in animals and humans, it has been classified as a Group 1 carcinogen in humans. Preserving food products with Sorghum bicolor sheath can minimise the [...] Read more.
Aflatoxin B1 (AFB1) is a recalcitrant metabolite produced by fungi species, and due to its intoxications in animals and humans, it has been classified as a Group 1 carcinogen in humans. Preserving food products with Sorghum bicolor sheath can minimise the contamination of agricultural products and avert ill health occasioned by exposure to AFB1. The current study investigated the ameliorating effect of Sorghum bicolor sheath hydrophobic extract (SBE-HP) enriched in Apigenin (API) on the hepatorenal tissues of rats exposed to AFB1. The SBE-HP was characterised using TLC and LC-MS and was found to be enriched in Apigenin and its methylated analogues. The study used adult male rats divided into four experimental cohorts co-treated with AFB1 (50 µg/kg) and SBE-HP (5 and 10 mg/kg) for 28 days. Biochemical, enzyme-linked immunosorbent assays (ELISA) and histological staining were used to examine biomarkers of hepatorenal function, oxidative status, inflammation and apoptosis, and hepatorenal tissue histo-architectural alterations. Data were analysed using GraphPad Prism 8.3.0, an independent t-test, and a one-way analysis of variance. Co-treatment with SBE-HP ameliorated an upsurge in the biomarkers of hepatorenal functionality in the sera of rats, reduced the alterations in redox balance, resolved inflammation, inhibited apoptosis, and preserved the histological features of the liver and kidney of rats exposed to AFB1. SBE-HP-containing API is an excellent antioxidant regiment. It can amply prevent the induction of oxidative stress, inflammation, and apoptosis in the hepatorenal system of rats. Therefore, supplementing animal feeds and human foods with SBE-HP enriched in Apigenin may reduce the burden of AFB1 intoxication in developing countries with a shortage of effective antifungal agents. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Metabolism of AFB1. Following chronic exposure to AFB1, AFB1 is rapidly absorbed into the enterocytes by passive diffusion and is absorbed into the liver for metabolism by arrays of CYP isoforms. Specifically, NADPH-dependent reductase converts AFB1 into aflatoxicol (AFL), CYP3A4 and CYP1A2 convert AFB1 into aflatoxin Q1 (AFQ1), CYP1A1 and CYP1A2 convert AFB1 into aflatoxin M1 (AFM1), or aflatoxin P1 (AFP1) through an O-demethylation reaction. These intermediates are clinically irrelevant as they are not heavily implicated in mutagenicity, carcinogenicity, and teratogenicity. The hepatic expression of CYP1A1, CYP1A2, CYP2E6, CPY3A4, and CYP3A5 is associated with AFBO formation, a clinically relevant intermediate, implicated in AFB<sub>1</sub> toxicity and carcinogenicity. The mechanisms of AFBO toxicity are through the formation of protein adducts, DNA adducts, and lipid peroxidation. Specifically, epoxide hydrolase at PH 7.4 hydrolyses AFBO to AFB<sub>1</sub> 8,9-dihydrodiol, which can form a Schiff’s base with primary amino groups in lysine residues, forming a protein adduct with lysine. This adduct is known to inhibit protein synthesis. In addition, AFBO binds to the guanine of DNA and forms a DNA adduct known as 8,9-dihydro-8-(N<sup>7</sup>-guanyl)-9-hydroxy-AFB1, leading to double-strand break, point mutation, activation of pro-oncogenes, and suppression of tumour suppressor genes. There is a safe pathway (blue arrow) for degrading AFBO, mediated by GST, an enzyme that mediates the conjugation of AFBO and GSH to form the AFB1-<span class="html-italic">S</span>-G complex. MRP binds to this complex and removes them from the hepatocytes for excretion in the bile and urine. <span class="html-italic">Created by ChemDraw</span>.</p> Full article ">Figure 2
<p>Characterisation of the key components in SBE-HP using TLC and LC-MS analyses. (<b>A</b>) TLC analysis on a normal phase silica plate, eluted with CH<sub>2</sub>Cl<sub>2</sub>/MeOH 12:1, revealed that SBE-HP separated into three clusters with <span class="html-italic">RF</span>s of 0.27, 0.83, and approx. 1. (<b>B</b>) LC-MS traces of key compounds in SBE-HP. Negative ion mode was presented for clarity.</p> Full article ">Figure 3
<p>Experimental protocol of AFB<sub>1</sub> and API-rich hydrophobic fraction of <span class="html-italic">S. bicolor</span> extracts 28 consecutive days. (<b>A</b>) In vivo screening of the hepatoprotective effect of SBE-HP on AFB1-challenged adult male Wistar Albino rats for 28 consecutive days, (<b>B</b>) effect of SBE-HP on the mean body weight change of AFB1-treated treated, and (<b>C</b>) Kaplan-Meier curve of rats treated with SBE-HP and AFB<sub>1</sub>. Created by <a href="https://app.biorender.com/" target="_blank">https://app.biorender.com/</a> (accessed on 8 July 2022).</p> Full article ">Figure 4
<p>Effect of SBE-HP on the liver and kidney function of rats treated with AFB<sub>1</sub> for 28 d. Experimental doses: AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 suggests the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher quantity; ALT: Alanine aminotransferase; AST: aspartate aminotransferase, ALP: Alkaline phosphatase; GGT: gamma-glutamyl transferase.</p> Full article ">Figure 5
<p>Effect of SBE-HP on the tissue concentrations of CAT, SOD, and GPx in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 indicates the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose; SOD: Superoxide dismutase; CAT: Catalase; GPx: Glutathione peroxidase.</p> Full article ">Figure 6
<p>Effect of SBE-HP on the tissue concentrations of GST, GSH, and TSH in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 suggests the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose; GST: Glutathione S-transferase; GSH: reduced glutathione; TSH: Total sulfhydryl group.</p> Full article ">Figure 7
<p>Effect of SBE-HP on the concentrations of LPO and RONS in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 suggests the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose; LPO: Lipid peroxidation; RONS: Reactive oxygen and nitrogen species.</p> Full article ">Figure 8
<p>Effect of SBE-HP on the tissue concentrations of XO, MPO, and NO in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 indicates the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose; XO: Xanthine oxidase; MPO: Myeloperoxidase; NO: Nitric oxide.</p> Full article ">Figure 9
<p>Effect of SBE-HP on tissue concentrations of IL-1β and IL-10 in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 suggests the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose; IL-1β: Interleukin-1beta; IL-10: Interleukin-10.</p> Full article ">Figure 10
<p>Effect of SBE-HP on tissue concentrations of caspase-9 and caspase-3 in the liver and kidney of rats treated with AFB<sub>1</sub> for 28 d. AFB<sub>1</sub> at 50 μg/kg; SBE-HP-D1 at 5 mg/kg; SBE-HP-D2 at 10 mg/kg. Values are expressed as mean ± SD for six rats per treatment cohort. Connecting lines indicate groups compared to one another. The significance level was set at (<span class="html-italic">p</span> &lt; 0.05); <span class="html-italic">p</span> &lt; 0.05 suggests the level of significance; <span class="html-italic">p</span> &gt; 0.05: not significant. AFB<sub>1</sub>: Aflatoxin B1; D1: lower dose; D2: higher dose.</p> Full article ">Figure 11
<p>(<b>a</b>) Control Plates of the kidney show a focal area of mild congestion and apoptotic bodies with typical tissue architecture. AFB<sub>1</sub> alone plates show disseminated glomerular messangialisation (thin arrow) and the extent of infiltration of the cortex by inflammatory cells (bold arrowhead). SBE-HP alone plate (not shown) tissues appear normal and relatively like those from control tissue sections. AFB<sub>1</sub> with SBE-HP-D1 and SBE-HP-D2 plates dose-dependently improved histo-architecture of the kidney with the mild presence of inflammatory cells. (<b>b</b>) AFB<sub>1</sub> alone shows areas of focal congestion (bold arrows), infiltration of zone 2 by inflammatory cells, mild hydropic/ballooning degeneration of the hepatocytes, and moderate microvesicular steatosis (tiny arrows). SBE-HP alone (plate not shown) tissue morphologies are similar to the control plate. AFB<sub>1</sub> with SBE-HP-D1 and SBE-HP-D2 plates improved hepatic cytoarchitecture with mild focal congestion and infiltration of zone 2 by inflammatory cells. H and E-stained sections; magnification at ×400.</p> Full article ">Figure 12
<p>Proposed mechanism of SBE-HP ameliorative effect on AFB<sub>1</sub>-mediated toxicities in the liver and kidney of an experimental rat model. SBE-HP prevented AFB<sub>1</sub>-induced oxidative and nitrosative stress and inflammation by reducing the activity of CYP1A2, NF-kB-mediated generation of pro-inflammatory cytokines, and IL-1β. SBE-HP also reduced apoptosis by altering the Bcl-2/Bax ratio favouring caspase 9 and caspase 3 activity. Created by <a href="https://app.biorender.com/" target="_blank">https://app.biorender.com/</a> (accessed on 9 July 2022).</p> Full article ">
13 pages, 2163 KiB  
Article
Effect of Thermal Processes on S-Allyl Cysteine Content in Black Garlic
by Kanokwan Manoonphol, Uthaiwan Suttisansanee, Chadamas Promkum and Chaniphun Butryee
Foods 2023, 12(6), 1227; https://doi.org/10.3390/foods12061227 - 13 Mar 2023
Cited by 2 | Viewed by 3051
Abstract
As a key component of black garlic (BG) products, S-allyl cysteine (SAC) is useful in reducing oxidative stress and inflammation. Several BG products with a high SAC content have been developed by thermal processing; however, the optimum conditions for thermal treatment for producing [...] Read more.
As a key component of black garlic (BG) products, S-allyl cysteine (SAC) is useful in reducing oxidative stress and inflammation. Several BG products with a high SAC content have been developed by thermal processing; however, the optimum conditions for thermal treatment for producing Thai garlic (multicloves) with a high SAC content compared to Chinese garlic (single clove) are still unknown. Moreover, the mechanism underlying the increase in SAC content in BG is unclear. Thus, this study aimed to investigate the optimum thermal condition for developing Thai BG with high SAC content base on methods A (70 °C and 80% RH) and B (60–75 °C and 80–85% RH). The total phenolic contents and antioxidant activities of Thai fresh garlic, Thai BG, and their powder forms were also compared. Method A worked the best for both types of garlic. The results indicated that the SAC content increased significantly after 7 days of fermentation and decreased drastically afterward with prolonged heat treatment. The optimum thermal condition for producing Thai fresh garlic and Chinese fresh garlic with high SAC content was 70 °C and a relative humidity of 80% for 12 days in an industrial fermentation chamber. The SAC content of Thai BG and Chinese BG increased approximately 139- and 122-fold, respectively. Furthermore, significant antioxidant capabilities determined by ferric ion-reducing antioxidant power, 2,2-diphenyl-1-picrylhydrazyl radical scavenging, and oxygen radical absorbance capacity assays were 34-, 6-, and 3-fold higher, respectively, than those of fresh garlic. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Full article ">Figure 1
<p>Changes in the color of Thai fresh garlic strain and Chinese garlic strain during period of thermal processes methods; (<b>A</b>) temperature was set at 70 °C, 80% RH, 42 days and (<b>B</b>) temperature was set at Step 1: 75 °C, 85% RH for 9 days. Step 2: 70 °C, 80% RH for 1 days; Step 3: 60 °C, 80% RH for 21 days.</p> Full article ">
19 pages, 1551 KiB  
Article
Thymus Species from Romanian Spontaneous Flora as Promising Source of Phenolic Secondary Metabolites with Health-Related Benefits
by Mihai Babotă, Oleg Frumuzachi, Alexandru Nicolescu, Maria Inês Dias, José Pinela, Lillian Barros, Mikel Añibarro-Ortega, Dejan Stojković, Tamara Carević, Andrei Mocan, Víctor López and Gianina Crișan
Antioxidants 2023, 12(2), 390; https://doi.org/10.3390/antiox12020390 - 6 Feb 2023
Cited by 3 | Viewed by 2802
Abstract
Wild thyme aerial parts (Serpylli herba) are recognized as a valuable herbal product with antioxidant, anti-inflammatory, and antibacterial effects. Although pharmacopoeial regulations allow its collection exclusively from Thymus serpyllum, substitution with other species is frequent in current practice. This study [...] Read more.
Wild thyme aerial parts (Serpylli herba) are recognized as a valuable herbal product with antioxidant, anti-inflammatory, and antibacterial effects. Although pharmacopoeial regulations allow its collection exclusively from Thymus serpyllum, substitution with other species is frequent in current practice. This study analyzed the phenolic composition, antioxidant, and enzyme-inhibitory and antimicrobial activity of the hydroethanolic extracts obtained from five Romanian wild thyme species (Thymus alpestris, T. glabrescens, T. panonicus, T. pulcherimus and T. pulegioides). The analysis of individual phenolic constituents was performed through LC-ESI-DAD/MS2, while for the in vitro evaluation of antioxidant potential, TEAC, FRAP, DPPH, TBARS and OxHLIA assays were employed. The anti-enzymatic potential was tested in vitro against tyrosinase, α-glucosidase and acetylcholinesterase. High rosmarinic acid contents were quantified in all species (20.06 ± 0.32–80.49 ± 0.001 mg/g dry extract); phenolic acids derivatives (including salvianolic acids) were confirmed as the principal metabolites of T. alpestris and T. glabrescens, while eriodictyol-O-di-hexoside was found exclusively in T. alpestris. All species showed strong antioxidant potential and moderate anti-enzymatic effect against α-glucosidase and acetylcholinesterase, showing no anti-tyrosinase activity. This is the first detailed report on the chemical and biological profile of T. alpestris collected from Romanian spontaneous flora. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>TPC and TFC values measured for the <span class="html-italic">Thymus</span> species studied. The results are expressed as average ± standard deviation of three parallel measurements. Statistical differences were assessed by one-way ANOVA, followed by Tukey’s HSD post hoc test (α = 0.05). Different lower case letters indicate significant differences between extracts. <b>T. alp</b>—<span class="html-italic">T. alpestris</span>, <b>T. glb</b>—<span class="html-italic">T. glabrescens</span>, <b>T. pan</b>—<span class="html-italic">T. pannonicus</span>, <b>T. plc</b>—<span class="html-italic">T. pulegioides</span>, <b>T. plg</b>—<span class="html-italic">T. pulcherimus</span>.</p> Full article ">Figure 2
<p>Results of the in vitro antioxidant capacity of <span class="html-italic">Thymus</span> species studied through DPPH, FRAP, TEAC, OxHLIA and TBARS assays. The results are expressed as average ± standard deviation of three parallel measurements. Statistical differences were assessed by one-way ANOVA, followed by Tukey’s HSD post hoc test (α = 0.05). Different lower-case letters indicate significant differences between extracts. <b>T. alp</b>—<span class="html-italic">T. alpestris</span>, <b>T. glb</b>—<span class="html-italic">T. glabrescens</span>, <b>T. pan</b>—<span class="html-italic">T. pannonicus</span>, <b>T. plc</b>—<span class="html-italic">T. pulegioides</span>, <b>T. plg</b>—<span class="html-italic">T. pulcherimus</span>.</p> Full article ">Figure 3
<p>The concentration (expressed as logC, in μg/mL)-dependent antioxidant activity of <span class="html-italic">Thymus</span> extracts and gallic acid against superoxide radical generated by the xanthine/xanthine oxidase method. <b>T. alp</b>—<span class="html-italic">T. alpestris</span>, <b>T. glb</b>—<span class="html-italic">T. glabrescens</span>, <b>T. pan</b>—<span class="html-italic">T. pannonicus</span>, <b>T. plc</b>—<span class="html-italic">T. pulegioides</span>, <b>T. plg</b>—<span class="html-italic">T. pulcherimus</span>.</p> Full article ">Figure 4
<p>The concentration (expressed as logC, in μg/mL)-dependent enzyme-inhibitory activity of <span class="html-italic">Thymus</span> extracts against <span class="html-italic">α</span>-glucosidase (<b>A</b>) and acetylcholinesterase (AChE) (<b>B</b>). Acarbose and galantamine were used as reference substances in anti-<span class="html-italic">α</span>-glucosidase and anti-AChE assays, respectively. <b>T. alp</b>—<span class="html-italic">T. alpestris</span>, <b>T. glb</b>—<span class="html-italic">T. glabrescens</span>, <b>T. pan</b>—<span class="html-italic">T. pannonicus</span>, <b>T. plc</b>—<span class="html-italic">T. pulegioides</span>, <b>T. plg</b>—<span class="html-italic">T. pulcherimus</span>.</p> Full article ">
17 pages, 1681 KiB  
Article
Identification of Small-Molecule Bioactive Constituents from the Leaves of Vaccinium bracteatum Confirms It as a Potential Functional Food with Health Benefits
by Yin-Yin Wang, Jun-Sheng Zhang, Xin-Xin Wang, Lin-Lin Tian, Yu-Peng Li, Chao Wang, Ren-Fen Ma, Yi-Ke Yin, Jie Bao and Hua Zhang
Foods 2023, 12(1), 177; https://doi.org/10.3390/foods12010177 - 1 Jan 2023
Cited by 5 | Viewed by 1802
Abstract
The health benefits of Vaccinium bracteatum are well recorded in ancient Chinese medical books and were also demonstrated by modern researches. However, the relationship between its beneficial functions and specific chemical constituents has not been fully characterized. This study investigated the bioactive small-molecule [...] Read more.
The health benefits of Vaccinium bracteatum are well recorded in ancient Chinese medical books and were also demonstrated by modern researches. However, the relationship between its beneficial functions and specific chemical constituents has not been fully characterized. This study investigated the bioactive small-molecule constituents in the leaves of V. bracteatum, which afforded 32 compounds including ten new ones (1–9) and ten pairs of enantiomers (9–18). Their structures with absolute configurations were elucidated by spectroscopic methods, especially nuclear magnetic resonance (NMR) and electronic circular dichroism (ECD) analyses, with 1–4 bearing a novel revolving-door shaped scaffold. While half-compounds exhibited decent antioxidant activity by scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, all except 19 and 20 exerted significant capturing activity against diammonium 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) radicals. In addition, the new iridoids 1, 5, 6, and 7 exerted apparent neuroprotective activity toward PC12 cells, with 1 being comparable to the positive control, and selective compounds also displayed anti-diabetic and anti-inflammatory properties by inhibiting α-glucosidase and NO production, respectively. The current work revealed that the bioactive small-molecule constituents could be closely related to the functional food property of the title species. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Structures of the chemical constituents from <span class="html-italic">V. bracteatum</span>.</p> Full article ">Figure 2
<p>Key 2D NMR correlations for <b>1</b>, <b>5</b>, <b>5r</b>, and <b>9</b> (for a better illustration, the enantiomers of <b>1</b> and <b>5r</b> were used to show the NOE/NOESY signals).</p> Full article ">Figure 3
<p>Experimental and calculated ECD spectra for <b>7</b>, <b>9</b>–<b>18</b>, and <b>21</b>.</p> Full article ">Figure 4
<p>Neuroprotective evaluation of iridoids <b>1</b>–<b>8</b> in PC12 cells. (<b>A</b>) The cell viability of PC12 cells upon 6-OHDA treatment in the concentration range of 50–250 μM; (<b>B</b>) neuroprotective effects of <b>1</b>–<b>8</b> at 50 μM against 6-OHDA-induced PC12 cell injury. Data are expressed as mean ± SD from triplicate experiments. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, compared with the control group.</p> Full article ">
15 pages, 2273 KiB  
Article
Bioactive Compounds and Antioxidant Properties of Wild Rocket (Diplotaxis Tenuifolia L.) Grown under Different Plastic Films and with Different UV-B Radiation Postharvest Treatments
by Raffaele Romano, Fabiana Pizzolongo, Lucia De Luca, Eugenio Cozzolino, Massimo Rippa, Lucia Ottaiano, Pasquale Mormile, Mauro Mori and Ida Di Mola
Foods 2022, 11(24), 4093; https://doi.org/10.3390/foods11244093 - 17 Dec 2022
Cited by 4 | Viewed by 2116
Abstract
Rocket species are rich in nutrients with well-known bioactive activity, but their content depends on several factors, such as plant–UV radiation interaction. In this work, we measured the production of nutritional elements in wild rocket (Diplotaxis tenuifolia L.) leaves as a function [...] Read more.
Rocket species are rich in nutrients with well-known bioactive activity, but their content depends on several factors, such as plant–UV radiation interaction. In this work, we measured the production of nutritional elements in wild rocket (Diplotaxis tenuifolia L.) leaves as a function of exposure to UV-B radiation by adopting a combined approach. The wild rocket plants were grown under three greenhouse cover films (A, B, and C) having different transmittivity to UV-B and the fresh-cut leaves were exposed to UV-B in postharvest for 45, 150, 330, and 660 s. The content of chlorophyll, carotenoids, phenolic compounds, ascorbic acid, and the antioxidant activity were determined. Chlorophyll, carotenoids, and total phenolic content were significantly increased by the combination of Film C and treatment with UV-B for 45 s. The predominant phenolic compounds were kaempferol, isorhamnetin, and quercetin. Film C also elicited an increase in ascorbic acid (the most abundant antioxidant compound in the range 374–1199 per 100 g of dry matter) and antioxidant activity. These findings highlighted an increase in bioactive compound content in the wild rocket when it was cultivated under Film C (diffused light film with a tailored UV-B transmission dose) and treated with UV-B radiation for 45 s postharvest, corresponding to an energy dose of 0.2 KJ m−2. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Dry matter percentage (% <span class="html-italic">w</span>/<span class="html-italic">w</span>) in wild rocket leaves as affected by greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05); ns indicates no significant differences.</p> Full article ">Figure 2
<p>Chlorophyll a content (mg/100 g of dry weight D.W.) in wild rocket leaves as affected by interaction between greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window) and UV-B applications (I: not treated with UV-B; II: treated with UV-B for 45 s; III: treated with UV-B for 150 s; IV: treated with UV-B for 330 s; V: treated with UV-B for 660 s). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p> Full article ">Figure 3
<p>Chlorophyll b content (mg/100 g of dry weight D.W.) in wild rocket leaves as affected by interaction between greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window) and UV-B applications (I: not treated with UV-B; II: treated with UV-B for 45 s; III: treated with UV-B for 150 s; IV: treated with UV-B for 330 s; V: treated with UV-B for 660 s). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p> Full article ">Figure 4
<p>Carotenoid content (mg/100 g of dry weight D.W.) in wild rocket leaves as affected by interaction between greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window) and UV-B applications (I: not treated with UV-B; II: treated with UV-B for 45 s; III: treated with UV-B for 150 s; IV: treated with UV-B for 330 s; V: treated with UV-B for 660 s). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p> Full article ">Figure 5
<p>Total phenolic content (mg gallic acid equivalent GAE/100 g of dry weight D.W.) in wild rocket leaves as affected by interaction between greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window) and UV-B applications (I: not treated with UV-B; II: treated with UV-B for 45 s; III: treated with UV-B for 150 s; IV: treated with UV-B for 330 s; V: treated with UV-B for 660 s). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p> Full article ">Figure 6
<p>Ascorbic acid content (mg/100 g of dry weight D.W.) in wild rocket leaves as affected by the interaction between greenhouse cover film (Film A: diffused light film; Film B: clear film; Film C: diffused light film with UV-B window) and UV-B application (I: not treated with UV-B; II: treated with UV-B for 45 s; III: treated with UV-B for 150 s; IV: treated with UV-B for 330 s; V: treated with UV-B for 660 s). Vertical bars indicate standard error; different letters indicate significant differences according to Tukey’s test (<span class="html-italic">p</span> ≤ 0.05).</p> Full article ">
18 pages, 2290 KiB  
Article
Protective Effect of Foxtail Millet Protein Hydrolysate on Ethanol and Pyloric Ligation-Induced Gastric Ulcers in Mice
by Bowei Zhang, Xiaoxiao Rao, Yunhui Zhang, Weijia Dai, Yingchuan Xu, Congying Zhao, Zhenjia Chen, Jin Wang, Dancai Fan, Xiaowen Wang and Shuo Wang
Antioxidants 2022, 11(12), 2459; https://doi.org/10.3390/antiox11122459 - 14 Dec 2022
Cited by 8 | Viewed by 2408
Abstract
Foxtail millet has been traditionally considered to possess gastroprotective effects, but studies evaluating its use as a treatment for gastric ulcers are lacking. Here, we assessed the antiulcer effects of foxtail millet protein hydrolysate (FPH) and explored its mechanism by using blocking agents. [...] Read more.
Foxtail millet has been traditionally considered to possess gastroprotective effects, but studies evaluating its use as a treatment for gastric ulcers are lacking. Here, we assessed the antiulcer effects of foxtail millet protein hydrolysate (FPH) and explored its mechanism by using blocking agents. In a mouse model of ethanol-induced gastric ulcers, pretreatment with FPH reduced the ulcerative lesion index, downregulated the expression of inflammatory cytokines in the gastric tissue, increased the activity of antioxidant enzymes, and improved the oxidative status. FPH increased constitutive the activity of nitric oxide synthase (cNOS), NO levels, and mucin expression in gastric mucosa, and inhibited the activation of the ET-1/PI3K/Akt pathway. In a mouse model of pyloric ligation-induced gastric ulcers, FPH inhibited gastric acid secretion and decreased the activity of gastric protease. Pretreatment of mice with the sulfhydryl blocker NEM and the NO synthesis inhibitor L-NAME abolished the gastroprotective effect of FPH, but not the KATP channel blocker glibenclamide and the PGE2 synthesis blocker indomethacin. Among the peptides identified in FPH, 10 peptides were predicted to have regulatory effects on the gastric mucosa, and the key sequences were GP and PG. The results confirmed the gastroprotective effect of FPH and revealed that its mechanism was through the regulation of gastric mucosal mucus and NO synthesis. This study supports the health effects of a millet-enriched diet and provides a basis for millet protein as a functional food to improve gastric ulcers and its related oxidative stress. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

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<p>Gastroprotective effects of FPH on ethanol-induced gastric ulcers in mice. Protocol of the animal experiments (<b>A</b>). Macroscopic evaluation of gastric mucosal lesions (<b>B</b>). Ulcerative lesion index and gastric MPO activity (<b>C</b>). Levels of inflammatory cytokines in the gastric tissue (<b>D</b>). Data were expressed as the mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparison tests (<span class="html-italic">n</span> = 10) was used to analyze the significance. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001 represent significant differences; ns, not significant.</p> Full article ">Figure 2
<p>Effects of FPH on gastric histopathology, defensive factors, and antioxidant enzyme activity. HE staining of gastric mucosa sections. The arrows marked with “a” show mucosal epithelial defects, arrows marked with “b” show inflammatory spillover, arrows marked with “c” show glandular disorder, and arrows marked with “d” show submucosal edema (40× magnification, bar = 200 μm). (<b>A</b>). NO, PGE2, and mucin content in the gastric tissues (<b>B</b>). Antioxidant enzyme activities in gastric tissues (<b>C</b>). Antioxidant enzyme activities in the gastric tissues. Data were expressed as the mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparison tests (<span class="html-italic">n</span> = 10) was used to analyze the significance. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **** <span class="html-italic">p</span> &lt; 0.0001 represent significant differences; ns, not significant.</p> Full article ">Figure 3
<p>Effects of FPH on gastric acid and digestive enzyme secretion in mice with pyloric ligation-induced gastric ulcers. Protocol of the animal experiments (<b>A</b>). Gastric secretion parameters (<b>B</b>). Activity of pepsin in the gastric juice (<b>C</b>). Duodenal activity of digestive enzymes (<b>D</b>). Data are expressed as the mean ± SEM. one-way ANOVA followed by Tukey’s multiple comparison tests (<span class="html-italic">n</span> = 10) was used to analyze the significance. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001 represent significant differences; ns, not significant.</p> Full article ">Figure 4
<p>Effects of molecular pathway blockers on the gastroprotective effect of FPH. Protocol of the animal experiments (<b>A</b>). Ulcerative lesion index (ULI) (<b>B</b>). ULI inhibition (<b>C</b>). Mucin content in gastric tissues after the NEM treatment (<b>D</b>). NO content in gastric tissues after the NEM treatment (<b>E</b>). Data are expressed as the mean ± SEM. Two-way ANOVA followed by Bonferroni’s multiple comparison tests (<span class="html-italic">n</span> = 10) was used to test significance for (<b>B</b>,<b>D</b>,<b>E</b>). One-way ANOVA followed by Tukey’s multiple comparison tests (<span class="html-italic">n</span> = 10) was used to analyze the significance for (<b>C</b>). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **** <span class="html-italic">p</span> &lt; 0.0001 represent significant differences; ns, not significant.</p> Full article ">Figure 5
<p>Molecular mechanisms of the gastroprotective effects of FPH on ethanol-induced gastric ulcers. Activity of NOS (<b>A</b>). Expression of eNOS (<b>B</b>). Expression of ET-1, PI3K, and Akt (<b>C</b>,<b>D</b>). mRNA expression of MUC5AC, MUC1, and MUC6 (<b>E</b>). Data are expressed as the mean ± SEM. One-way ANOVA followed by Tukey’s multiple comparison tests (<span class="html-italic">n</span> = 10 for (<b>A</b>,<b>E</b>); <span class="html-italic">n</span> = 4 for (<b>B</b>,<b>C</b>)) was used to analyze the significance. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001 represent significant differences; ns, not significant.</p> Full article ">
28 pages, 2482 KiB  
Article
Antioxidative and Immunomodulating Properties of Aronia melanocarpa Extract Rich in Anthocyanins
by Kseniya Bushmeleva, Alexandra Vyshtakalyuk, Dmitriy Terenzhev, Timur Belov, Evgeny Nikitin and Vladimir Zobov
Plants 2022, 11(23), 3333; https://doi.org/10.3390/plants11233333 - 1 Dec 2022
Cited by 12 | Viewed by 2046
Abstract
The fruits of Aronia melanocarpa are well known due to their high anthocyanin content that may be effective in preventing certain health disorders arising from oxidative stress. Various polyphenolic compounds such as anthocyanins and flavonoids are responsible for the multiple effects of chokeberry. [...] Read more.
The fruits of Aronia melanocarpa are well known due to their high anthocyanin content that may be effective in preventing certain health disorders arising from oxidative stress. Various polyphenolic compounds such as anthocyanins and flavonoids are responsible for the multiple effects of chokeberry. The aim of this study was to determine in vitro how active the black chokeberry anthocyanins are in scavenging radicals and to evaluate in vivo their immunomodulating capacity. Using the method of column chromatography, we extracted the anthocyanins of black chokeberries, i.e., cyanidin-3-O-galactoside with a purity of over 93.7%. Using HPLC and spectrophotometric analysis, the flavonoid content was determined. Following the analysis of the tests with AAPH and DPPH, the chokeberry cyanidin-3-O-galactoside was found much better than individual anthocyanins in regard to antioxidant capacity. The range of concentrations was revealed, showing the protective effect of anthocyanins on the RPMI-1788 cell culture against cyclophosphamide, as well as against osmotic and peroxide hemolysis. An immunomodulating effect on the functional activity of phagocytes was revealed in vivo as a result of oral administration of chokeberry cyanidin-3-O-galactoside and a mixture composed of cyanidin-3-O-glucoside and cyanidin-3-O-galactoside standards. Consequently, anthocyanins, in particular cyanidin-3-O-galactoside, play an important role, demonstrating immunomodulating effects when chokeberries are consumed. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Chemiluminescent intensity attenuation (<b>A</b>) (TAR—total antioxidant reactivity) and time (<b>B</b>) (TRAP—total reactive antioxidant potential) vs. the concentration of the studied compounds. Values obtained from the quenching of luminol-enhanced chemiluminescence (CL).</p> Full article ">Figure 2
<p>Osmotic (<b>A</b>) and peroxide (<b>B</b>) red blood cell hemolysis intensity vs. the concentration of cyanidin-3-O-galactoside Aronia extract, cyanidin-3-O-galactoside and cyanidin-3-O-glucoside using the model system of osmotic red blood cell damage and oxidative damage caused by Fenton reagent. Values are expressed as percent inhibition (mean ± S.D.).</p> Full article ">Figure 3
<p>Cytoprotective activity of anthocyanins regarding the lymphoblastic RPMI-1788 cell line.</p> Full article ">Figure 4
<p>Main hematology results for whole blood of lab animals. The corresponding baselines for each group on the 1st day of the experiment are taken as 100%. (<b>A</b>) White blood cell (WBC) count; (<b>B</b>) red blood cell (RBC) count; (<b>C</b>) hemoglobin (HGB) concentration; (<b>D</b>) platelet (PLT) count; (<b>E</b>) mean platelet volume (MPV). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 4 Cont.
<p>Main hematology results for whole blood of lab animals. The corresponding baselines for each group on the 1st day of the experiment are taken as 100%. (<b>A</b>) White blood cell (WBC) count; (<b>B</b>) red blood cell (RBC) count; (<b>C</b>) hemoglobin (HGB) concentration; (<b>D</b>) platelet (PLT) count; (<b>E</b>) mean platelet volume (MPV). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 5
<p>Leukocyte profile. The corresponding initial values for each group in the 1st day of the experiment are accepted as 100%. (<b>A</b>) Lymphocyte (LYM) count; (<b>B</b>) ratio of lymphocytes to total leukocytes (LYM%); (<b>C</b>) monocyte (MON) count; (<b>D</b>) ratio of monocytes to total leukocytes (MON%); (<b>E</b>) neutrophil granulocyte (GRA) count; (<b>F</b>) ratio of granulocytes to total leukocytes (GRA%). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 5 Cont.
<p>Leukocyte profile. The corresponding initial values for each group in the 1st day of the experiment are accepted as 100%. (<b>A</b>) Lymphocyte (LYM) count; (<b>B</b>) ratio of lymphocytes to total leukocytes (LYM%); (<b>C</b>) monocyte (MON) count; (<b>D</b>) ratio of monocytes to total leukocytes (MON%); (<b>E</b>) neutrophil granulocyte (GRA) count; (<b>F</b>) ratio of granulocytes to total leukocytes (GRA%). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 6
<p>Phagocytic activity of neutrophil granulocytes and rat peripheral blood monocytes. Values (phagocytic index (<b>A</b>,<b>B</b>), phagocyte number (<b>C</b>,<b>D</b>)) are expressed in conventional units (c.u.); phagocytic activity (<b>E</b>,<b>F</b>) is expressed in %; phagocytosis completion index (<b>G</b>,<b>H</b>)) is expressed in c.u. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, significance of differences compared to the 1st day of the experiment; ° <span class="html-italic">p</span> &lt; 0.05, °° <span class="html-italic">p</span> &lt; 0.01, significance of differences between the animal groups as compared to the Control Group.</p> Full article ">Figure 6 Cont.
<p>Phagocytic activity of neutrophil granulocytes and rat peripheral blood monocytes. Values (phagocytic index (<b>A</b>,<b>B</b>), phagocyte number (<b>C</b>,<b>D</b>)) are expressed in conventional units (c.u.); phagocytic activity (<b>E</b>,<b>F</b>) is expressed in %; phagocytosis completion index (<b>G</b>,<b>H</b>)) is expressed in c.u. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, significance of differences compared to the 1st day of the experiment; ° <span class="html-italic">p</span> &lt; 0.05, °° <span class="html-italic">p</span> &lt; 0.01, significance of differences between the animal groups as compared to the Control Group.</p> Full article ">Figure 7
<p>Spontaneous and zymosan-activated CL of rat neutrophils: (<b>A</b>)—CL intensity (I<sub>max</sub>, c.u.); (<b>B</b>)—area under the CL curve (AUC, c.u.); (<b>C</b>)—the time showing peak CL (T<sub>max</sub>, c.u.); (<b>D</b>)—activation index (I<sub>act</sub>, c.u.). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 7 Cont.
<p>Spontaneous and zymosan-activated CL of rat neutrophils: (<b>A</b>)—CL intensity (I<sub>max</sub>, c.u.); (<b>B</b>)—area under the CL curve (AUC, c.u.); (<b>C</b>)—the time showing peak CL (T<sub>max</sub>, c.u.); (<b>D</b>)—activation index (I<sub>act</sub>, c.u.). * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of the experiment.</p> Full article ">Figure 8
<p>Malondialdehyde (MDA) level of peripheral blood of rats. * <span class="html-italic">p</span> &lt; 0.05, significance of differences in animal groups compared to the 1st day of experiment. ° <span class="html-italic">p</span> &lt; 0.05, significance of differences between animal groups compared to Control Group.</p> Full article ">
19 pages, 2737 KiB  
Article
By-Product of the Red Ginseng Manufacturing Process as Potential Material for Use as Cosmetics: Chemical Profiling and In Vitro Antioxidant and Whitening Activities
by Hui-E Zhang, Meng-Yao Chu, Tao Jiang, Xin-Hong Song, Jian-Feng Hou, Li-Ye Cheng, Ye Feng, Chang-Bao Chen and En-Peng Wang
Molecules 2022, 27(23), 8202; https://doi.org/10.3390/molecules27238202 - 24 Nov 2022
Cited by 6 | Viewed by 2299
Abstract
Red ginseng (RG), which is obtained from heated Panax ginseng and is produced by steaming followed by drying, is a valuable herb in Asian countries. Steamed ginseng dew (SGD) is a by-product produced in processing red ginseng. In the present study, phytochemical profiling [...] Read more.
Red ginseng (RG), which is obtained from heated Panax ginseng and is produced by steaming followed by drying, is a valuable herb in Asian countries. Steamed ginseng dew (SGD) is a by-product produced in processing red ginseng. In the present study, phytochemical profiling of extracts of red ginseng and steamed ginseng dew was carried out using gas chromatography-mass spectrometry (GC-MS) and rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) analysis. Additionally, antioxidant activities (DPPH, ·OH, and ABTS scavenging ability) and whitening activities (tyrosinase and elastase inhibitory activity) were analyzed. Phytochemical profiling revealed the presence of 66 and 28 compounds that were non-saponin components in chloroform extracts of red ginseng and steamed ginseng dew (RG-CE and SGD-CE), respectively. Meanwhile, there were 20 ginsenosides identified in n-butanol extracts of red ginseng and steamed ginseng dew (RG-NBE and SGD-NBE). By comparing the different polar extracts of red ginseng and steamed ginseng dew, it was found that the ethyl acetate extract of red ginseng (RG-EAE) had the best antioxidant capacity and whitening effect, the water extract of steamed ginseng dew (SGD-WE) had stronger antioxidant capacity, and the SGD-NBE and SGD-CE had a better whitening effect. This study shows that RG and SGD have tremendous potential to be used in the cosmetic industries. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract
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<p>Antioxidant activity of RG and SGD extracts. (<b>A</b>) Hydroxyl radical scavenging activity of RG extracts. (<b>B</b>) Hydroxyl radical scavenging activity of SGD extracts. RG-FDP, freeze-dried powder of red ginseng; RG-CE, chloroform extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; RG-NBE, n-butanol extract of red ginseng; RG-WE, water extract of red ginseng; SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew; vitamin C, positive control. Different groups indicate significant difference according to ANOVA followed by Tukey’s T-test(<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Comparative analysis of three antioxidant activities of RG and SGD extracts by radar chart. RG-FDP, freeze-dried powder of red ginseng; RG-CE, chloroform extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; RG-NBE, n-butanol extract of red ginseng; RG-WE, water extract of red ginseng; SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew. (Blue symbols indicate hydroxyl radical scavenging activity, green symbols indicate DPPH free radical scavenging ability, and red symbols indicate ABTS radical scavenging activity. <span class="html-italic">n</span> = 3).</p> Full article ">Figure 3
<p>Antioxidant activity of RG and SGD extracts. (<b>A</b>) DPPH free radical scavenging ability of RG and its extracts. (<b>B</b>) DPPH free radical scavenging ability of SGD extracts and its extracts. RG-FDP, freeze-dried powder of red ginseng; RG-CE, chloroform extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; RG-NBE, n-butanol extract of red ginseng; RG-WE, water extract of red ginseng; SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew; vitamin C, positive control. Different groups indicate significant difference according to ANOVA followed by Tukey’s T-test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Antioxidant activity of RG and SGD extracts. (<b>A</b>) ABTS radical scavenging activity of RG extracts. (<b>B</b>) ABTS radical scavenging activity of SGD extracts. RG-FDP, freeze-dried powder of red ginseng; RG-CE, chloroform extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; RG-NBE, n-butanol extract of red ginseng; RG-WE, water extract of red ginseng; SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew; vitamin C, positive control. Different groups indicate significant difference according to ANOVA followed by Tukey’s T-test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Inhibitory activity of samples on tyrosinase. (<b>A</b>) Tyrosine inhibition rate of RG extracts. (<b>B</b>) Tyrosine inhibition rate of SGD extracts. RG-CE, chloroform extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; KA, kojic acid (positive control); SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew.</p> Full article ">Figure 6
<p>Changes in IC<sub>50</sub> values on tyrosinase and elastase activities. (<b>A</b>) The IC<sub>50</sub> values of RG and SGD extract on tyrosinase activity. (<b>B</b>) The IC<sub>50</sub> values of RG-NBE and SGD-NBE on elastase activity. RG-CE, chloroform extract of red ginseng; RG-NBE, n-butanol extract of red ginseng; RG-EAE, ethyl acetate extract of red ginseng; SGD-FDP, freeze-dried powder of steamed ginseng dew; SGD-CE, chloroform extract of steamed ginseng dew; SGD-EAE, ethyl acetate extract of steamed ginseng dew; SGD-NBE, n-butanol extract of steamed ginseng dew; SGD-WE, water extract of steamed ginseng dew.</p> Full article ">Figure 7
<p>Structure and secondary mass spectrum of Rb1. (<b>A</b>) Structure diagram. (<b>B</b>) Secondary mass spectrometry.</p> Full article ">Figure 8
<p>TIC of RG-CE and SGD-CE with GC-MS.</p> Full article ">Figure 9
<p>Venn diagram of similarities and differences of volatile compounds in RG-CE and SGD-CE.</p> Full article ">
11 pages, 3046 KiB  
Article
Impact of Green Extraction on Curcuminoid Content, Antioxidant Activities and Anti-Cancer Efficiency (In Vitro) from Turmeric Rhizomes (Curcuma longa L.)
by Kanjana Singh, Somdet Srichairatanakool, Teera Chewonarin, Adchara Prommaban, Rajnibhas Sukeaw Samakradhamrongthai, Margaret Anne Brennan, Charles Stephen Brennan and Niramon Utama-ang
Foods 2022, 11(22), 3633; https://doi.org/10.3390/foods11223633 - 14 Nov 2022
Cited by 19 | Viewed by 3279
Abstract
Turmeric (Curcuma longa L.) powder is widely used as a spice and seasoning in Asian countries. This study investigated the effect of turmeric extracts on the anticancer activity of Huh7 and HCT 116 cells. The curcumin bioactive compounds were extracted using various [...] Read more.
Turmeric (Curcuma longa L.) powder is widely used as a spice and seasoning in Asian countries. This study investigated the effect of turmeric extracts on the anticancer activity of Huh7 and HCT 116 cells. The curcumin bioactive compounds were extracted using various methods such as microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE) and traditional extraction (TDE). The yield of dried extracts from MAE was found to be the highest at 17.89%, followed by UAE and TDE, with 11.34% and 5.54%, respectively. Antioxidant activities such as TPC, DPPH and FRAP from MAE were higher than those of UAE and TDE. The total curcuminoid contents from the novel extractions were higher than those from traditional extraction methods. For instance, curcuminoid contents from MAE, UAE and TDE were 326.79, 241.17 and 215.83 mg/g, respectively. Due to having the highest bioactive compounds and extraction yield, turmeric extract from MAE was used to investigate the potential anticancer properties. The extract showed significant cytotoxic potential against the human liver (Huh7) and human colon (HCT116) cell lines, in concentrations ranging from 31.25 to 1000.00 µg/mL. Turmeric extracts using MAE have potential anticancer effects on Huh7 and HCT116 cells. This study serves as scientific data for the chemotherapeutic properties of turmeric extracts and their use as functional ingredients. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Chromatogram of turmeric extracts using HPLC and chemical structure of curcumin.</p> Full article ">Figure 2
<p>Scanning electron micrograph of turmeric extracts using UAE (<b>a</b>), MAE (<b>b</b>) and TDE (<b>c</b>).</p> Full article ">Figure 3
<p>The viability of Huh7 (<b>A</b>,<b>B</b>) and HCT116 (<b>C</b>,<b>D</b>) cancer cells treated with standard curcumin and turmeric extracts at the indicated concentrations for 24 h and 48 h were determined using an MTT assay <sup>a–b</sup> (<span class="html-italic">p</span> &lt; 0.001).</p> Full article ">Figure 4
<p>Direct microscope observations of Huh7 cancer cells treated with turmeric extracts at different concentrations ((<b>A</b>); 0, (<b>B</b>); 31.25, (<b>C</b>); 62.50, (<b>D</b>); 125.00, (<b>E</b>); 250.00, (<b>F</b>); 500.00 and (<b>G</b>); 1000.00 µg/mL) under a microscope with 10× magnification compared with untreated control (<b>A</b>); red arrows present the morphology change of death cells.</p> Full article ">
18 pages, 5799 KiB  
Article
Systematical Characterization of the Cotton Di19 Gene Family and the Role of GhDi19-3 and GhDi19-4 as Two Negative Regulators in Response to Salt Stress
by Lanjie Zhao, Youzhong Li, Yan Li, Wei Chen, Jinbo Yao, Shengtao Fang, Youjun Lv, Yongshan Zhang and Shouhong Zhu
Antioxidants 2022, 11(11), 2225; https://doi.org/10.3390/antiox11112225 - 11 Nov 2022
Cited by 5 | Viewed by 1946
Abstract
Drought-induced 19 (Di19) protein is a Cys2/His2 (C2H2) type zinc-finger protein, which plays a crucial role in plant development and in response to abiotic stress. This study systematically investigated the characteristics of the GhDi19 gene family, including the member number, gene structure, chromosomal [...] Read more.
Drought-induced 19 (Di19) protein is a Cys2/His2 (C2H2) type zinc-finger protein, which plays a crucial role in plant development and in response to abiotic stress. This study systematically investigated the characteristics of the GhDi19 gene family, including the member number, gene structure, chromosomal distribution, promoter cis-elements, and expression profiles. Transcriptomic analysis indicated that some GhDi19s were up-regulated under heat and salt stress. Particularly, two nuclear localized proteins, GhDi19-3 and GhDi19-4, were identified as being in potential salt stress responsive roles. GhDi19-3 and GhDi19-4 decreased sensitivity under salt stress through virus-induced gene silencing (VIGS), and showed significantly lower levels of H2O2, malondialdehyde (MDA), and peroxidase (POD) as well as significantly increased superoxide dismutase (SOD) activity. This suggested that their abilities were improved to effectively reduce the reactive oxygen species (ROS) damage. Furthermore, certain calcium signaling and abscisic acid (ABA)-responsive gene expression levels showed up- and down-regulation changes in target gene-silenced plants, suggesting that GhDi19-3 and GhDi19-4 were involved in calcium signaling and ABA signaling pathways in response to salt stress. In conclusion, GhDi19-3 and GhDi19-4, two negative transcription factors, were found to be responsive to salt stress through calcium signaling and ABA signaling pathways. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Phylogenic analysis of Di19 proteins in cotton and other plants. The neighbor-joining (NJ) phylogenetic tree was built with MEGA 7.0, and the bootstrap values from 1000 replicates are indicated at each branch. Each plant species was marked with different characters and colors. Di19 proteins of 11 plants were labeled with different colors and patterns.</p> Full article ">Figure 2
<p>Gene structure, motifs, domains, and protein 3D-structures analysis of 16 <span class="html-italic">GhDi19</span> genes in <span class="html-italic">G. hirsutum</span>. (<b>A</b>) Phylogenetic tree of GhDi19s. A neighbor-joining (NJ) phylogenetic tree was built using MEGA 7.0, and the bootstrap values of 1000 replicates were listed at each branch. (<b>B</b>) Structural analysis of 16 <span class="html-italic">GhDi19s</span>. (<b>C</b>) Motif of 16 GhDi19 proteins. Ten motifs were investigated using the MEME online tool. (<b>D</b>) Multiple sequence alignments of 16 GhDi19 proteins. Two zinc-finger domains and one NLS are marked by red and blue boxes, respectively. Di19_zinc-binding domain and the Di19_C terminal domain of Di19 proteins are underlined in black. (<b>E</b>–<b>H</b>) 3D structures of GhDi19-1, GhDi19-2, GhDi19-3, and GhDi19-4 proteins.</p> Full article ">Figure 3
<p>Conserved promoter cis-element and expression profile analysis of <span class="html-italic">GhDi19s</span>. (<b>A</b>) Phylogenetic tree of GhDi19s. A neighbor-joining (NJ) phylogenetic tree was built with MEGA 7.0, and the bootstrap values from 1000 replicates are indicated at each branch. (<b>B</b>) Conserved promoter analysis of GhDi19 genes. Different cis-elements in the 2000bp promoter region upstream of Di19 gene are marked with different color boxes. (<b>C</b>) RNA-sequence data analysis of <span class="html-italic">GhDi19s</span> under different abiotic stresses (cold, hot, PEG, and NaCl). Transcriptomic data normalization and visualization was performed. The color scale in the lower right corner of the heatmap represents the fragments per kilobase million (FPKM) values, which were standardized by log10.</p> Full article ">Figure 4
<p>Association network of Di19s. (<b>A</b>) Association network of Di19 proteins. A total of nine association protein signaling pathways were identified. The blue letters represent Di19 protein signaling pathway or other protein families. (<b>B</b>) Association network of GhDi19-3 and GhDi19-4 with other Di19 proteins. A total of ten association proteins were identified. The blue letters represent CDPK proteins and other proteins.</p> Full article ">Figure 5
<p>Subcellular localization of GhDi19-3 and GhDi19-4 in tobacco epidermal cells. 35S-GFP, GhDi19-3-GFP, and GhDi19-4-GFP represented confocal imaging of epidermal cells of transiently expression of the control GFP, GhDi19-3 and GhDi19-4, respectively. GFP fluorescence signals were mainly detected in nucleus of the tobacco epidermal cells. GFP, GFP fluorescence images; Bright, bright field image of the same leaf on the left; Merged, GFP fluorescence and bright merged image of the same leaf. Scale bars are 25 µm.</p> Full article ">Figure 6
<p>Silenced <span class="html-italic">GhDi19-3</span> and <span class="html-italic">GhDi19-4</span> improves the tolerance of cotton to salt stress. (<b>A</b>) The <span class="html-italic">GhDi19-3</span> and <span class="html-italic">GhDi19-4</span> silenced genes, control (TRV: 00) and positive control (TRV: GhCLA1) cotton plants before treatments. (<b>B</b>) Expression of <span class="html-italic">GhDi19-3</span> and <span class="html-italic">GhDi19-4</span> in TRV: 00, TRV: GhDi19-3, and TRV: GhDi19-4 cotton plants. (<b>C</b>) Phenotype of TRV: 00, TRV: GhDi19-3, and TRV: GhDi19-4 cotton plants under mock and treatment of 400 mM NaCl, H<sub>2</sub>O<sub>2</sub> contents (<b>D</b>), POD (<b>E</b>), MDA contents (<b>F</b>) and SOD activity (<b>G</b>) of the silenced genes and control plants (TRV: 00) under treatments. Asterisks indicate significant differences (independent <span class="html-italic">t</span>-tests): * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 7
<p>qRT-PCR analysis of calcium signaling related and ABA-responsive genes under simulated salt stress in the control and <span class="html-italic">GhDi19-3</span>- and <span class="html-italic">GhDi19-4</span>-silenced plants. Transcript levels of <span class="html-italic">GhCDPK2-1A</span>, <span class="html-italic">GhCDPK2-1D</span>, <span class="html-italic">GhCDPK2-2A</span>, <span class="html-italic">GhCDPK2-2D</span>, <span class="html-italic">GhCIPK11-A</span>, <span class="html-italic">GhCIPK11-D</span>, <span class="html-italic">GhABI5</span>, <span class="html-italic">GhRD29B-A</span>, and <span class="html-italic">GhRD29B-D</span> genes were determined by qRT-PCR, using <span class="html-italic">GhUBQ7</span> gene as the reference. Asterisks indicate significant differences (independent <span class="html-italic">t</span>-tests): * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 8
<p>A model describing the silenced <span class="html-italic">GhDi19-3</span> and <span class="html-italic">GhDi19-4</span> gene regulates cotton tolerance to salt stress. <span class="html-italic">GhDi19-3</span> and <span class="html-italic">GhDi19-4</span> were up-regulated under salt stress, and the silencing of <span class="html-italic">GhDi19-3</span> or <span class="html-italic">GhDi19-4</span> genes led to a change in up- and down-regulation of calcium signaling and ABA signaling pathway related genes (<span class="html-italic">GhCDPK2-1A</span>, <span class="html-italic">GhCDPK2-1D</span>, <span class="html-italic">GhCDPK2-2A</span>, <span class="html-italic">GhCDPK2-2D</span>, <span class="html-italic">GhCIPK11-A</span>, <span class="html-italic">GhCIPK11-D</span>, <span class="html-italic">GhABI5</span>, <span class="html-italic">GhRD29B-A</span>, and <span class="html-italic">GhRD29B-D</span>), which then resulted in their abilities improving to effectively reduce the ROS damage when cotton plants were subject to salt stress. Taken together, GhDi19-3 and GhDi19-4, as two negative regulatory transcription factors, responded to salt stress by being involved in calcium signaling and ABA signaling pathways.</p> Full article ">
19 pages, 4478 KiB  
Article
Protective Effect of Flavonoids from Mulberry Leaf on AAPH-Induced Oxidative Damage in Sheep Erythrocytes
by Qinhua Zheng, Weijian Tan, Xiaolin Feng, Kexin Feng, Wenting Zhong, Caiyu Liao, Yuntong Liu, Shangjian Li and Wenzhong Hu
Molecules 2022, 27(21), 7625; https://doi.org/10.3390/molecules27217625 - 7 Nov 2022
Cited by 21 | Viewed by 2578
Abstract
To evaluate the antioxidant activity of flavonoids extracted from Chinese herb mulberry leaves (ML), flavonoids from mulberry leaves (FML) were extracted and purified by using ultrasonic-assisted enzymatic extraction and D101 macroporous resin. Using LC-MS/MS-Liquid Chromatography with tandem mass spectrometry analysis, hesperidin, rutoside, hyperoside, [...] Read more.
To evaluate the antioxidant activity of flavonoids extracted from Chinese herb mulberry leaves (ML), flavonoids from mulberry leaves (FML) were extracted and purified by using ultrasonic-assisted enzymatic extraction and D101 macroporous resin. Using LC-MS/MS-Liquid Chromatography with tandem mass spectrometry analysis, hesperidin, rutoside, hyperoside, cyanidin-3-o-glucoside, myricitrin, cyanidin, and quercetin were identified, and NMR and UV were consistent with the verification of IR flavonoid characteristics. The antioxidant activity of FML has also been evaluated as well as the protective effect on 2,2 0-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress. The results showed that FML exhibited powerful antioxidant activity. Moreover, FML showed dose-dependent protection against AAPH-induced sheep erythrocytes’ oxidative hemolysis. In the enzymatic antioxidant system, pretreatment with high FML maintained the balance of SOD, CAT, and GSH-Px; in the non-enzymatic antioxidant system, the content of MDA can be effectively reduced after FML treatment. This study provides a research basis for the development of natural products from mulberry leaves. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The procedure for the extraction and purification of FML.</p> Full article ">Figure 2
<p>The procedure for the AAPH-treated erythrocyte hemolysis assay.</p> Full article ">Figure 3
<p>(<b>A</b>) UV scan spectrum of FML and rutin; (<b>B</b>) FT-IR spectrum of FML.</p> Full article ">Figure 4
<p>Total ion chromatogram (TIC) of FML in LC-MS/MS: (<b>A</b>) ESI-; (<b>B</b>) ESI+.</p> Full article ">Figure 5
<p>FML NMR hydrogen spectra (1H NMR). (Note: FML had six heterotopic hydrogen signals at δ7.43~7.59(part A), 6.77~7.05(part B), 6.19~6.42(part C), δ5.34~5.45(part D), 5.11~5.14(part E), and 3.95~4.00(part F) ppm in 1H-NMR).</p> Full article ">Figure 6
<p>FML NMR carbon spectra (13C NMR). (Note: The δ values of c2~c3 signals in both part A and part B of FML were concentrated between δ 114.95~126.09 and δ 145.29~148.70; Part C is the DMSO–D6).</p> Full article ">Figure 7
<p>Determination of the antioxidant activity of the five compounds: (<b>A</b>) DPPH· scavenging ability; (<b>B</b>) ·OH scavenging ability; (<b>C</b>) O<sub>2</sub><sup>−</sup> scavenging ability; (<b>D</b>) ABTS scavenging ability. Vc, vitamin C.</p> Full article ">Figure 8
<p>Inhibitory effects of various concentrations of FML on AAPH-induced erythrocyte hemolysis (Note: The same small letters indicate no significant difference between groups, different small letters indicate a significant difference between groups, <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 9
<p>Effect of FML on changes in MDA content (<b>A</b>) and enzyme activities of SOD (<b>B</b>), GPx (<b>C</b>), and CAT (<b>D</b>) in erythrocytes with AAPH-induced oxidative damage; (<b>E</b>) possible intracellular antioxidant mechanisms of FML in an AAPH-induced model of oxidative damage (Note: The same small letters indicate no significant difference between groups, different small letters indicate a significant difference between groups, <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 10
<p>SEM micrographs of sheep erythrocyte samples: (<b>A</b>) normal erythrocyte; (<b>B</b>) AAPH-treated erythrocyte; (<b>C</b>) erythrocyte preincubated with diosmetin before AAPH treatment.</p> Full article ">
17 pages, 1456 KiB  
Article
Unravelling Phytochemical and Bioactive Potential of Three Hypericum Species from Romanian Spontaneous Flora: H. alpigenum, H. perforatum and H. rochelii
by Mihai Babotă, Oleg Frumuzachi, Andrei Mocan, Mircea Tămaș, Maria Inês Dias, José Pinela, Dejan Stojković, Marina Soković, Alexandru Sabin Bădărău, Gianina Crișan, Lillian Barros and Ramona Păltinean
Plants 2022, 11(20), 2773; https://doi.org/10.3390/plants11202773 - 19 Oct 2022
Cited by 9 | Viewed by 2420
Abstract
Hypericum perforatum L., also known as St. John’s Wort, is recognized worldwide as a valuable medicinal herb; however, other Hypericum species were intensively studied for their bioactive potential. To fill the research gap that exists in the scientific literature, a comparative evaluation between [...] Read more.
Hypericum perforatum L., also known as St. John’s Wort, is recognized worldwide as a valuable medicinal herb; however, other Hypericum species were intensively studied for their bioactive potential. To fill the research gap that exists in the scientific literature, a comparative evaluation between H. alpigenum Kit., H. perforatum L. and H. rochelii Griseb. & Schenk was conducted in the present study. Two types of herbal preparations obtained from the aerial parts of these species were analyzed: extracts obtained through maceration and extracts obtained through magnetic-stirring-assisted extraction. LC-DAD-ESI-MSn analysis revealed the presence of phenolic acids, flavan-3-ols and flavonoid derivatives as the main constituents of the above-mentioned species. Moreover, all extracts were tested for their antioxidant, enzyme-inhibitory and antimicrobial potential. Our work emphasizes for the first time a detailed description of H. rochelii phenolic fractions, including their phytochemical and bioactive characterization. In comparison with the other two studied species, H. rochelii was found as a rich source of phenolic acids and myricetin derivatives, showing important antioxidant, anticholinesterase and antibacterial activity. The study offers new perspectives regarding the chemical and bioactive profile of the less-studied species H. alpigenum and H. rochelii. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1
<p>Enzyme inhibition curves of <span class="html-italic">Hypericum</span> extracts against acetylcholinesterase (AChE, (<b>A</b>–<b>C</b>)), <span class="html-italic">α</span>-glucosidase (<b>D</b>–<b>F</b>) and tyrosinase (<b>G</b>). <b>HAA</b> (<span class="html-italic">H. alpigenum</span> MSA extraction), <b>HAM</b> (<span class="html-italic">H. alpigenum</span> maceration), <b>HPA</b> (<span class="html-italic">H. perforatum</span> MSA extraction), <b>HPM</b> (<span class="html-italic">H. perforatum</span> maceration), <b>HRA</b> (<span class="html-italic">H. rochelii</span> MSA extraction), <b>HRM</b> (<span class="html-italic">H. rochelii</span> maceration).</p> Full article ">Figure 2
<p>Correlation plot (correlogram) that shows Pearson’s correlation coefficients between chemical constituents of analyzed extracts and total antioxidant capacity and enzyme-inhibitory activities of the extracts. Red color represents negative correlation (values from 0 to −1), which indicates an inverse proportional relationship between the compared variables, while the blue color represents positive correlation (values from 0 to +1), which indicates a direct proportional relationship between the compared variables. The e diameter of each dot and the intensity of the colors indicate the strongness of correlation (increased values of Pearson’s correlation coefficients). Asterisk (*) indicates the statistical significant results (<span class="html-italic">p</span>&lt;0.05).</p> Full article ">
17 pages, 2043 KiB  
Article
Changes in Phenolic Compounds and Antioxidant Activity during Development of ‘Qiangcuili’ and ‘Cuihongli’ Fruit
by Huifen Zhang, Jing Pu, Yan Tang, Miao Wang, Kun Tian, Yongqing Wang, Xian Luo and Qunxian Deng
Foods 2022, 11(20), 3198; https://doi.org/10.3390/foods11203198 - 13 Oct 2022
Cited by 23 | Viewed by 2719
Abstract
Plums are widely consumed, contain high levels of phenolic compounds, and have strong antioxidant activity. In this study, the main Sichuan cultivars ‘Qiangcuili’ and ‘Cuihongli’ were used to study the changes in the appearance, internal quality, phenolic compounds, and antioxidant activities during fruit [...] Read more.
Plums are widely consumed, contain high levels of phenolic compounds, and have strong antioxidant activity. In this study, the main Sichuan cultivars ‘Qiangcuili’ and ‘Cuihongli’ were used to study the changes in the appearance, internal quality, phenolic compounds, and antioxidant activities during fruit development and the expression of phenolic-compound-related structural genes. The results showed that, during development of the two plums, the total soluble solid and soluble sugar contents were highest at the mature stage. The phenolic contents (total phenol content (TPC), total flavonoid content (TFC), and total flavanol content (TFAC)) trended gradually downward as the fruits of the two cultivars matured, while the total anthocyanin content gradually increased in ‘Cuihongli’. The main phenolic components were neochlorogenic acid, chlorogenic acid, ferulic acid, benzoic acid, rutin, and proanthocyanidin B1. The changes in the DPPH and FRAP scavenging activities decreased with fruit ripening. The antioxidant capacity was positively correlated with the TPC, TFC, and TFAC. In the two cultivars, the total phenols, phenolic components, and antioxidant capacity were higher in the peel than in the pulp. CHS, PAL3, and HCT1 may be the regulatory genes related to the accumulation of phenolic substances in the pericarp and pulp of ‘Qiangcuili’ and ‘Cuihongli’. HCT1 may be an important regulator involved in the accumulation of chlorogenic acid in plums. The changes in the quality of the phenols, phenolic components, and antioxidant activity were elucidated during the development of the main plum cultivars in Sichuan, particularly the changes that provided a theoretical basis for the development of bioactive substances in local plum cultivars. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Changes in appearance quality during plum development. Photographs of the six development stages (S1–S6) of (<b>A</b>) ‘Qiangcuili’(QCL) and (<b>B</b>) ‘Cuihongli’(CHL); (<b>C</b>) fruit weight; (<b>D</b>) firmness; (<b>E</b>) fruit L* value; (<b>F</b>) fruit a* value; (<b>G</b>) fruit b* value. Developmental stages S1–S6 correspond to days 42, 56, 70, 84, 98, and 112 after anthesis. Different letters (a–f) indicate significant differences at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test. Each point on the graph shows the mean and standard error, n = 3.</p> Full article ">Figure 2
<p>Changes in TSS (<b>A</b>), soluble sugar contents (<b>B</b>), TA contents (<b>C</b>), and Vc contents (<b>D</b>) during the development of plums. QCL-P and CHL-P represent the peel of ‘Qiangcuili’ and ‘Cuihongli’, respectively. QCL-F and CHL-F represent the flesh of ‘Qiangcuili’ and ‘Cuihongli’, respectively, the same as below. Different letters (a–f) indicate significant differences at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test. Each point on the graph shows the mean and standard error, n = 3.</p> Full article ">Figure 3
<p>Total phenolic (TPC), total flavonoid (TFC), total flavanol (TFAC), and total anthocyanin contents (TMAC) in the peel and pulp of the plum cultivars ‘Qiangcuili’ and ‘Cuihongli’ during development. (<b>A</b>) TPC; (<b>B</b>) TFC; (<b>C</b>) TFAC; and (<b>D</b>) TMAC. Different letters (a–f) indicate significant differences at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test. Each point on the graph shows the mean and standard error, n = 3.</p> Full article ">Figure 4
<p>Change in antioxidant activity in the skins of plum cultivars ‘Qiangcuili’ and ‘Cuihongli’ during development. (<b>A</b>) DPPH radical scavenging capacity; (<b>B</b>) FRAP ferric reducing antioxidant capacity. Different letters (a–f) indicate significant differences at <span class="html-italic">p</span> &lt; 0.05 by Duncan’s test. Each point on the graph shows the mean and standard error, n = 3.</p> Full article ">Figure 5
<p>Gene expression of <span class="html-italic">PAL1</span>, <span class="html-italic">PAL3</span>, <span class="html-italic">C4H</span>, <span class="html-italic">4CL1</span>, <span class="html-italic">4CL2</span>, <span class="html-italic">CHS</span>, <span class="html-italic">HCT1</span>, <span class="html-italic">HCT2</span>, and <span class="html-italic">HCT3</span> during the development of QCL-P, CHL-P, QCL-F, and CHL-F.</p> Full article ">
17 pages, 1351 KiB  
Communication
Comparison of Phytochemical Profile and In Vitro Bioactivity of Beverages Based on the Unprocessed and Extruded Sesame (Sesamum indicum L.) Seed Byproduct
by María Fernanda Quintero-Soto, Ramona Julieta Espinoza-Moreno, Jennifer Vianey Félix-Medina, Fernando Salas-López, Cruz Fernando López-Carrera, Oscar Daniel Argüelles-López, Martha Elena Vazquez-Ontiveros and Mario Armando Gómez-Favela
Foods 2022, 11(20), 3175; https://doi.org/10.3390/foods11203175 - 12 Oct 2022
Cited by 2 | Viewed by 2142
Abstract
In this research functional beverages based on the unprocessed and extruded sesame seeds byproduct were fabricated; phytochemical profile, antioxidant, antidiabetic, and hypoglycemic potential were evaluated. Twenty-four phytochemical compounds were identified in total in both beverages; fourteen of the phytochemical compounds were not modified [...] Read more.
In this research functional beverages based on the unprocessed and extruded sesame seeds byproduct were fabricated; phytochemical profile, antioxidant, antidiabetic, and hypoglycemic potential were evaluated. Twenty-four phytochemical compounds were identified in total in both beverages; fourteen of the phytochemical compounds were not modified by the extrusion process. Seventeen of the 24 compounds were identified in the unprocessed sesame seeds byproduct flour beverage−10% (UB10) and 21 in the extruded sesame seeds byproduct flour beverage−10% (EB10). The compounds only identified in UB10 are caffeic acid, luteolin-7-O-glucoside, and isorhamnetin; and in EB10 those compounds were vanillic acid, acteoside, luteolin, quercetin, and melanoidins. No significant difference was observed in the content of total phenolic compounds (TPC) (14.90 and 15.97 mg GAE/100 mL) and total flavonoids (TF) (5.37 and 5.85 mg QE/100 mL). An increase in the biological activity of ESFB10 (IC50: ABTS = 0.19, DPPH = 0.21, α-amylase = 1.01, α-glucosidase = 0.17, DPP4 = 0.11 mg/mL) was observed, compared to UB10 (IC50: ABTS = 0.24, DPPH = 0.31, α-amylase = 2.29, α-glucosidase = 0.47, DPP4 = 0.30 mg/mL). Therefore, the extrusion process had a positive effect, which displayed the highest efficiency inhibiting the free radicals and enzymes related to carbohydrate metabolism. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Sensory acceptance analysis of extruded sesame seeds byproduct flour beverages. (<b>A</b>) Flavor; (<b>B</b>) Color; (<b>C</b>) Smell; (<b>D</b>) Global Acceptability. EB5: Beverage with 5% extruded sesame seeds byproduct flour; EB10: Beverage with 10% extruded sesame seeds byproduct flour; EB15: Beverage with 15% extruded sesame seeds byproduct flour; EB20: Beverage with 20% extruded sesame seeds byproduct flour; BE25: Beverage with 25% extruded sesame seeds byproduct flour; BE30: Beverage with 30% extruded sesame seeds byproduct flour. Different letters in the same graph indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) among the means according to the Fisher’s test.</p> Full article ">Figure 2
<p>Chromatography separation of phytochemical compounds found in extracts of (<b>A</b>) unprocessed sesame seeds byproduct flour beverage−10% (UB10) and (<b>B</b>) extruded sesame seeds byproduct flour beverage-10 (EB10). 1: 3-<span class="html-italic">O</span>-p-Coumaroylquinic acid, 2: Vanillic acid, 3: Caffeic acid, 4: Chlorogenic acid, 5: Syringic acid; 6: Acteoside, 7: Ferulic Acid, 8: Diosmetin, 9: Verbasoside, 10: 6-Methylquercetin-3-<span class="html-italic">O</span>-Rutinoside, 11: Crenatoside, 12: Quercetin-3-<span class="html-italic">O</span>-glucoside, 13: Aspartic acid + 4(hexoside), 14: 2(3-Deoxyglucosone) + 2(γ-aminobutyric acid), 15: Luteolin-7-<span class="html-italic">O</span>-glucoside, 16: Isorhamnetin-7-<span class="html-italic">O</span>-glucoside, 17: Salvigenin, 18: Phlorizin, 19: Quercetin, 20: Luteolin, 21: Genistein-7-<span class="html-italic">O</span>-galactoside, 22: Apigenin, 23: Isorhamnetin, 24: Arginine + sucrose derived compound.</p> Full article ">Figure 3
<p>A biplot based on principal component analysis of metabolites and bioactivities. Samples are represented by colored-shapes and the variables by black points. CQA: 3-<span class="html-italic">O</span>-p-Coumaroylquinic acid, VA: Vanillic acid, CaA: Caffeic acid, ChA: Chlorogenic acid, SA: Syringic acid; Act: Acteoside, FA: Ferulic Acid, Diosm: Diosmetin, Verb: Verbasoside, MGR: 6-Methylquercetin-3-<span class="html-italic">O</span>-Rutinoside, Cren: Crenatoside, QG: Quercetin-3-<span class="html-italic">O</span>-glucoside, M13: Aspartic acid + 4(hexoside), M14: 2(3-Deoxyglucosone) + 2(γ-aminobutyric acid), LutG: Luteolin-7-<span class="html-italic">O</span>-glucoside, IsoG: Isorhamnetin-7-<span class="html-italic">O</span>-glucoside, Salv: Salvigenin, Phl: Phlorizin, Quer: Quercetin, Lut: Luteolin, GenG: Genistein-7-<span class="html-italic">O</span>-galactoside, Apig: Apigenin, Isorh: Isorhamnetin, M24: Arginine + sucrose derived compound, TPC: Total Phenolic Compounds: TF: Total Flavonoids, AAM: <span class="html-italic">α</span>-amylase inhibition AG: <span class="html-italic">α</span>-glucosidase inhibition, DPP4: Dipeptidyl-peptidase-IV inhibition.</p> Full article ">
16 pages, 3442 KiB  
Article
Suppressive Effect of Fraxetin on Adipogenesis and Reactive Oxygen Species Production in 3T3-L1 Cells by Regulating MAPK Signaling Pathways
by Woonghee Lee, Gwonhwa Song and Hyocheol Bae
Antioxidants 2022, 11(10), 1893; https://doi.org/10.3390/antiox11101893 - 24 Sep 2022
Cited by 10 | Viewed by 2818
Abstract
Recent studies have identified obesity as one of the world’s most serious chronic disorders. Adipogenesis, in which preadipocytes are differentiated into mature adipocytes, has a decisive role in establishing the number of adipocytes and determining the lipid storage capacity of adipose tissue and [...] Read more.
Recent studies have identified obesity as one of the world’s most serious chronic disorders. Adipogenesis, in which preadipocytes are differentiated into mature adipocytes, has a decisive role in establishing the number of adipocytes and determining the lipid storage capacity of adipose tissue and fat mass in adults. Fat accumulation in obesity is implicated with elevated oxidative stress in adipocytes induced by reactive oxygen species (ROS). Adipogenesis regulation by inhibiting adipogenic differentiation and ROS production has been selected as the strategy to treat obesity. The conventional anti-obesity drugs allowed by the U.S. Food and Drug Administration have severe adverse effects. Therefore, various natural products have been developed as a solution for obesity, suppressing adipogenic differentiation. Fraxetin is a major component extracted from the stem barks of Fraxinus rhynchophylla, with various bioactivities, including anti-inflammatory, anticancer, antioxidant, and antibacterial functions. However, the effect of fraxetin on adipogenesis is still not clearly understood. We studied the pharmacological functions of fraxetin in suppressing lipid accumulation and its underlying molecular mechanisms involving 3T3-L1 preadipocytes. Moreover, increased ROS production induced by a mixture of insulin, dexamethasone, and 3-isobutylmethylxanthine (MDI) in 3T3-L1 was attenuated by fraxetin during adipogenesis. These effects were regulated by mitogen-activated protein kinase (MAPK) signaling pathways. Therefore, our findings imply that fraxetin possesses inhibitory roles in adipogenesis and can be a potential anti-obesity drug. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Impact of fraxetin on the formation of lipid droplets and proliferation of 3T3-L1 cells. (<b>A</b>,<b>B</b>) Nondifferentiated cells and mature adipocytes were incubated with different doses of fraxetin (0, 5, 10, 20, 50, and 100 µM). Relative lipid accumulation stained by oil red O was measured by a microplate reader at 490 nm of optical density. Orlistat (20 µM) was treated as a positive control. Scale bar: 50 µm. The different colors in bar graph represent the different concentration of fraxetin. Black bar indicates orlistat. (<b>C</b>) Cell proliferation analysis was conducted in 3T3-L1 preadipocytes. All data were obtained in triplicate. The symbols “*” and “**” represent significant differences between the differentiated group and others (*<span class="html-italic"> p </span>&lt; 0.05, **<span class="html-italic"> p </span>&lt; 0.01). “##” means considerable differences between the nondifferentiated group and others (##<span class="html-italic"> p </span>&lt; 0.01), as verified using one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 2
<p>Effect of fraxetin on the expression levels of adipogenesis-related proteins and genes in 3T3-L1 cells. (<b>A</b>,<b>B</b>) Protein expression levels of PPARγ and C/EBPα were assessed with Western blot analysis. The 3T3-L1 cells were incubated with or without fraxetin (20, 50, and 100 µM) for differentiation. Then, the cells were lysed, and a Western blot was performed. The protein levels of PPARγ and C/EBPα were normalized with β-actin. The different colors in bar graph represent the different concentration of fraxetin. (<b>C</b>) The transcriptional expressions of Pparg, Cebpa, Fabp4, Fasn, and Srebf1 genes were determined by RT-qPCR. GAPDH was used for the normalization of each gene expression. All experiments were performed at least three times. The asterisk symbol represents significant differences between the differentiated group and others (*<span class="html-italic"> p </span>&lt; 0.05, **<span class="html-italic"> p </span>&lt; 0.01). The crosshatch mark represents significant differences between the nondifferentiated group and others (#<span class="html-italic"> p </span>&lt; 0.05, ##<span class="html-italic"> p </span>&lt; 0.01, ###<span class="html-italic"> p </span>&lt; 0.001), as verified using unpaired t-tests or one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 3
<p>Effect of fraxetin on various stages of adipogenesis in 3T3-L1 cells. (<b>A</b>) Schematic diagram of the treatment period of fraxetin (100 µM) in confluent 3T3-L1 cells during adipogenesis. (<b>B</b>,<b>C</b>) The nondifferentiated cell and differentiated cells treated with fraxetin (100 µM) at different periods of adipogenesis were stained with oil red O solution on day 8. Relative lipid droplets eluted by isopropanol were quantified by measuring the absorbance at 490 nm. Scale bar: 50 µm. (<b>D</b>) Transcriptional expression of the <span class="html-italic">Klf5</span> gene was analyzed by RT-qPCR. All data were obtained in triplicate. The asterisk symbol represents significant differences between the MDI-treated group and others (**<span class="html-italic"> p </span>&lt; 0.01, ***<span class="html-italic"> p </span>&lt; 0.001). The crosshatch symbol represents significant differences between the nondifferentiated group and others (#<span class="html-italic"> p </span>&lt; 0.05, ###<span class="html-italic"> p </span>&lt; 0.001), as verified using unpaired <span class="html-italic">t</span>-tests or one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 4
<p>Fraxetin triggers cell-cycle shifts without apoptotic cell death. (<b>A</b>) Preadipocytes were incubated with diverse doses of fraxetin in an MDI induction medium for 2 days. After fixation, cells were stained by PI-retaining ribonuclease A and evaluated using flow cytometry. Each value in the histogram was determined by counting 10,000 events. The ratio of cells at each phase was represented in each histogram. (<b>B</b>) The impact of fraxetin with respect to apoptosis in MDI-treated cells was evaluated with Annexin V and PI double-staining assay. Confluent preadipocytes were incubated with different concentrations of fraxetin in an MDI induction medium for 2 days. Cells were treated by PI and Annexin V and assessed by flow cytometry. The number of cells in each dot plot was 10,000. All data were obtained in triplicate. The crosshatch mark indicates significant differences among the nondifferentiated group and others (#<span class="html-italic"> p </span>&lt; 0.05). The asterisk symbol represents significant differences between the MDI-treated group and others (*<span class="html-italic"> p </span>&lt; 0.05), as verified using one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 5
<p>Effect of fraxetin on ROS production and the transcriptional expression of scavenging genes during the adipogenesis of 3T3-L1 cells. (<b>A</b>) ROS production was evaluated with DCFH-DA using flow cytometry. Relative ROS production is represented as purple shadow in a bar graph. The different colors in bar graph represent the different concentration of fraxetin. (<b>B</b>) The mRNA expressions of scavenging genes (<span class="html-italic">Sod1</span>, <span class="html-italic">Sod2</span>) were analyzed with RT-qPCR. Each gene was normalized with the GAPDH gene. All data were obtained in triplicate. The asterisk indicates significant differences between the MDI-treated group and others (***<span class="html-italic"> p </span>&lt; 0.001). The crosshatch symbol represents significant differences between the nondifferentiated group and others (#<span class="html-italic"> p </span>&lt; 0.05, ##<span class="html-italic"> p </span>&lt; 0.01), as verified using unpaired <span class="html-italic">t</span>-tests or one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 6
<p>Effect of fraxetin on the regulation of adipogenesis through MAPK signaling. The protein expressions of the MAPK signaling pathway, including (<b>A</b>) p38, (<b>B</b>) Erk1/2, and (<b>C</b>) Jnk phosphorylation, were analyzed with Western blotting. Each phosphorylation level was normalized with each total protein level. All data were obtained in triplicate. The asterisk symbol represents significant differences between the MDI-treated group and others (*<span class="html-italic"> p </span>&lt; 0.05, **<span class="html-italic"> p </span>&lt; 0.01). The crosshatch symbol represents meaningful differences among the nondifferentiated group and others (##<span class="html-italic"> p </span>&lt; 0.01, ###<span class="html-italic"> p </span>&lt; 0.001), as verified using one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 7
<p>Effect of the combination of fraxetin and MAPK inhibitors on lipid accumulation. Fraxetin was cotreated with MAPK signaling inhibitors including SB203580, U0126, and SP600125 in the presence of an MDI induction medium. Then, the formation of lipid droplets was evaluated with oil red O solution. Relative staining rates were assessed by measuring the optical density at 490 nm. Scale bar: 50 µm. The red and black color represent the treatment with fraxetin and orlistat in the presence of MDI medium, respectively. The orange, blue, and purple colors in bar graph indicate the treatment with SB203580, U0126, and SP600125 in the presence of fraxetin, respectively. Experiments were performed at least three times. The crosshatch represents significant differences between the nondifferentiated group and others (###<span class="html-italic"> p </span>&lt; 0.001). The asterisk symbol represents significant differences between the MDI-treated group and others (*<span class="html-italic"> p </span>&lt; 0.05, **<span class="html-italic"> p </span>&lt; 0.01, ***<span class="html-italic"> p </span>&lt; 0.001). The symbol “a” <span class="html-italic">(p </span>&lt; 0.05) and “aa” <span class="html-italic">(p </span>&lt; 0.01) represent the significant effect of combination treatment between the fraxetin and MAPK-inhibitor-treated groups, as verified using one-way analysis of variance (ANOVA), followed by Dunnett’s post hoc analysis.</p> Full article ">Figure 8
<p>Schematic illustration of fraxetin on 3T3-L1 cells.</p> Full article ">
14 pages, 887 KiB  
Article
Biological Activity of Bark Extracts from Northern Red Oak (Quercus rubra L.): An Antioxidant, Antimicrobial and Enzymatic Inhibitory Evaluation
by Corneliu Tanase, Alexandru Nicolescu, Adrian Nisca, Ruxandra Ștefănescu, Mihai Babotă, Anca Delia Mare, Cristina Nicoleta Ciurea and Adrian Man
Plants 2022, 11(18), 2357; https://doi.org/10.3390/plants11182357 - 9 Sep 2022
Cited by 23 | Viewed by 4398
Abstract
The northern red oak (Quercus rubra L.) is an ornamental oak species native to eastern America, being an invasive species in Europe, with increasing coverage. The aim of this work was to evaluate the biological potential of red oak bark extracts. Aqueous [...] Read more.
The northern red oak (Quercus rubra L.) is an ornamental oak species native to eastern America, being an invasive species in Europe, with increasing coverage. The aim of this work was to evaluate the biological potential of red oak bark extracts. Aqueous and ethanolic preparations were obtained by two extraction methods: ultrasonic-assisted extraction (UAE) and microwave assisted extraction (MAE). The total phenolic and tannin contents were measured using spectrophotometric methods. The antioxidant activity was evaluated by two complementary methods (DPPH and ABTS). Antimicrobial potential was tested against five bacteria and three Candida species, and the effect on biofilm formation and synergism with gentamicin was also evaluated. Finally, enzyme inhibitory properties were assessed for α-glucosidase, tyrosinase, and acetylcholinesterase. The results indicated a higher phenolic content for the extracts obtained through MAE, while UAE bark extracts were rich in tannins. All the extracts exhibited antioxidant, anti-glucosidase, and anti-tyrosinase activity, while the antibacterial potential was mostly observed for the MAE extracts, especially against S. aureus, C. parapsilopsis, and C. krusei; inhibition of biofilm formation was observed only for MRSA. These findings show that the red oak bark might be an important source of bioactive compounds with antioxidant and antimicrobial properties. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Pearson’s correlation coefficients among TPC, TTC, DPPH, and ABTS calculated for the extracts obtained by microwave-assisted extraction (<b>A</b>) and by ultrasound-assisted extraction (<b>B</b>).</p> Full article ">Figure 2
<p>Graphs showing the dependence between the logC (of the concentration in terms of μg/mL) and the inhibition percentage for the ROBE samples in the case of α-glucosidase and tyrosinase inhibitory activity.</p> Full article ">
19 pages, 1420 KiB  
Article
Cellular Antioxidant Effect of an Aronia Extract and Its Polyphenolic Fractions Enriched in Proanthocyanidins, Phenolic Acids, and Anthocyanins
by Cécile Dufour, Jose A. Villa-Rodriguez, Christophe Furger, Jacob Lessard-Lord, Camille Gironde, Mylène Rigal, Ashraf Badr, Yves Desjardins and Denis Guyonnet
Antioxidants 2022, 11(8), 1561; https://doi.org/10.3390/antiox11081561 - 12 Aug 2022
Cited by 25 | Viewed by 3222
Abstract
Oxidative stress and chronic inflammation contribute to some chronic diseases. Aronia berries are rich in polyphenols. The aim of the present study was to characterize the cellular antioxidant effect of an aronia extract to reflect the potential physiological in vivo effect. Cellular in [...] Read more.
Oxidative stress and chronic inflammation contribute to some chronic diseases. Aronia berries are rich in polyphenols. The aim of the present study was to characterize the cellular antioxidant effect of an aronia extract to reflect the potential physiological in vivo effect. Cellular in vitro assays in three cell lines (Caco-2, HepG2, and SH-SY5Y) were used to measure the antioxidant effect of AE, in three enriched polyphenolic fractions (A1: anthocyanins and phenolic acids; A2: oligomeric proanthocyanidins; A3: polymeric proanthocyanidins), pure polyphenols and microbial metabolites. Both direct (intracellular and membrane radical scavenging, catalase-like effect) and indirect (NRF2/ARE) antioxidant effects were assessed. AE exerted an intracellular free radical scavenging activity in the three cell lines, and A2 and A3 fractions showed a higher effect in HepG2 and Caco-2 cells. AE also exhibited a catalase-like activity, with the A3 fraction having a significant higher activity. Only A1 fraction activated the NRF2/ARE pathway. Quercetin and caffeic acid are the most potent antioxidant polyphenols, whereas cyanidin and 5-(3′,4′-dihydroxyphenyl)-γ-valerolactone showed the highest antioxidant effect among polyphenol metabolites. AE rich in polyphenols possesses broad cellular antioxidant effects, and proanthocyanidins are major contributors. Polyphenol metabolites may contribute to the overall antioxidant effect of such extract in vivo. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Polyphenolic characterization of AE and the three polyphenolic fractions (A1, A2, and A3). Values are expressed in milligrams of polyphenols per gram of extract or fraction.</p> Full article ">Figure 2
<p>Dose–response curves of AE assessed with the AOP1 assay on different cell lines. HepG2 (panel <b>A</b>), SH-SY5Y (panel <b>B</b>) and Caco-2 cells (panel <b>C</b>) were treated for 4 h with AE in dose–response mode. Left column: kinetic profile of raw data. The raw data (RFUs, relative fluorescence units) were plotted in a kinetics-like mode showing the effect of light flashes on the fluorescence level. Data points, mean RFUs of triplicates; bars, SD. Central column: same data after normalization. Antioxidant indices correspond to control AUC minus data AUCs. Right column: antioxidant-index-based dose–response and sigmoid fit curve used for log EC<sub>50</sub> estimations (µg/mL). Data points correspond to measured antioxidant indexes. EC<sub>50</sub>, efficacy concentration which leads to a half-maximal response; R<sup>2</sup>, coefficient of determination obtained by non-linear regression model fit.</p> Full article ">Figure 3
<p>NRF2/ARE-mediated pathway activation by the fraction A1 in stable ARE-driven luciferase expression HepG2 cells. Cells were treated for 17 h with increasing doses of AE and fractions A1, A2, and A3. Only the A1 fraction showed a dose–response effect. On the left side, results are presented in gene expression fold increase compared with the control (cell culture medium only, gene expression = 1). Values under 1 indicate cytotoxicity effect. The bars are SD for mean relative luminescence units of duplicate. On the right side, the same results are presented as a dose–response curve (EC<sub>50</sub>: 742 µg/mL, 95% CI: [582.5, 944.9]). Data points correspond to the fold increase for each concentration and bars are SD. EC<sub>50</sub> is Efficacy Concentration which leads to half-maximal response. R<sup>2</sup> is the coefficient of determination obtained by non-linear regression model fit.</p> Full article ">
15 pages, 3018 KiB  
Article
Effects of a Whole Plant Extract of Scutellaria rubropunctata var. rubropunctata on Bone Metabolism with Estrogen Receptor Activation
by Misaki Watanabe, Tadahiro Yahagi, Takahiro Shirayama, Katsunori Miyake, Hitoshi Kotani, Takuya Ogawa and Keiichi Matsuzaki
Plants 2022, 11(16), 2075; https://doi.org/10.3390/plants11162075 - 9 Aug 2022
Cited by 2 | Viewed by 2369
Abstract
We screened natural resources for estrogen receptor (ER)-activating and bone metabolism-promoting activities with the aim of finding potential treatments for osteoporosis. A screen of 1531 extracts from Ryukyu Arc plants using a luciferase reporter assay identified an 80% MeOH extract of Scutellaria rubropunctata [...] Read more.
We screened natural resources for estrogen receptor (ER)-activating and bone metabolism-promoting activities with the aim of finding potential treatments for osteoporosis. A screen of 1531 extracts from Ryukyu Arc plants using a luciferase reporter assay identified an 80% MeOH extract of Scutellaria rubropunctata var. rubropunctata (SRE) with dose-dependent ER transcription-promoting activity. Importantly, SRE had no proliferative effect on human breast cancer cells. SRE enhanced the ALP activity of pre-osteoblast MC3T3-E1 cells after 72 h in culture and slightly enhanced mineralization at 14 and 21 d. SRE did not significantly affect the TRAP activity of RAW264.7 cells. Gene expression analysis in MC3T3-E1 cells by quantitative real-time PCR revealed that SRE upregulated the mRNA levels of Runx2, Osterix (Osx), Osteopontin (Opn), Osteocalcin (Ocn), Smad1, Smad4, and Smad5 at 72 h, and those of Runx2, Osx, Smad1, Smad4, and Smad5 at 21 d of osteogenic induction. Analysis of the expression levels of osteogenic markers suggested that SRE may promote osteogenic differentiation by acting at the early stage of differentiation rather than at the late stage of differentiation. These results indicate that SRE activates ER and induces osteoblast differentiation by activating Runx2 and Osx through the BMP/Smad pathway, suggesting that SRE may be useful for the prevention and treatment of postmenopausal osteoporosis. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1
<p>Estrogen receptor (ER) transcription-promoting activity of an 80% MeOH extract of <span class="html-italic">Scutellaria rubropunctata</span> var. <span class="html-italic">rubropunctata</span> (SRE). The ER transcription-promoting activity of SRE was examined using a luciferase reporter assay in HEK293 cells. The control (cont.), which contained vehicle only (DMSO), was set to 1. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 2
<p>Effect of the ER antagonist ICI 182, 780 on the ER transcription-promoting activity of SRE. The activity was assessed using a luciferase reporter assay in HEK293 cells. The control (first column) contained vehicle only (DMSO). Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 3
<p>SRE has no proliferation-promoting effect on human breast carcinoma cells. The proliferation of MCF7 cells was assessed using the MTT assay. The control contained DMSO only. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 4
<p>Effect of SRE on (<b>a</b>) alkaline phosphatase (ALP) activity and (<b>b</b>) the mRNA expression level of <span class="html-italic">A</span><span class="html-italic">lpl</span> in the MC3T3-E1 mouse osteoblastic cell line. The “non” sample was incubated without differentiation inducer but with vehicle (DMSO), and the control (cont.) was incubated with a differentiation medium and DMSO. The mRNA expression level of <span class="html-italic">Alpl</span> was measured by quantitative real-time PCR and normalized to the level of the reference gene <span class="html-italic">β-actin</span>. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 5
<p>Effect of SRE on mineralization in MC3T3-E1 cells incubated for 14 d. The “non” sample was incubated in a growth medium with DMSO, and “cont.” was incubated in a differentiation medium with DMSO. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 6
<p>Effect of SRE on mineralization in MC3T3-E1 cells incubated for 21 d. The “non” sample was incubated in a growth medium with DMSO, and the cont. sample was incubated in a differentiation medium with DMSO. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 7
<p>Effect of incubation with SRE for 72 h on the mRNA expression of osteoblast-differentiation-related genes in MC3T3-E1 cells. The mRNA expression levels of (<b>a</b>) <span class="html-italic">Runx2</span>, (<b>b</b>) <span class="html-italic">Osterix</span>, (<b>c</b>) <span class="html-italic">Osteopontin</span>, (<b>d</b>) <span class="html-italic">Osteocalcin</span>, (<b>e</b>) <span class="html-italic">Smad1</span>, (<b>f</b>) <span class="html-italic">Smad4</span>, and (<b>g</b>) <span class="html-italic">Smad5</span> were measured by quantitative real-time PCR and normalized to the levels of the reference gene <span class="html-italic">β-actin</span>. The “non” sample was incubated in an ordinary medium with DMSO, and the “cont.” sample” was incubated in a differentiation medium with DMSO. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 8
<p>Effect of incubation with SRE for 21 d on the mRNA expression of osteoblast-differentiation-related genes in MC3T3-E1 cells. The mRNA expression levels of (<b>a</b>) <span class="html-italic">Runx2</span>, (<b>b</b>) <span class="html-italic">Osterix</span>, (<b>c</b>) <span class="html-italic">Osteopontin</span>, (<b>d</b>) <span class="html-italic">Osteocalcin</span>, (<b>e</b>) <span class="html-italic">Smad1</span>, (<b>f</b>) <span class="html-italic">Smad4</span>, and (<b>g</b>) <span class="html-italic">Smad5</span> were measured by quantitative real-time PCR and normalized to the levels of the reference gene <span class="html-italic">β-actin</span>. The “non” sample was incubated in an ordinary medium with DMSO, and the “cont.” sample was incubated in a differentiation medium with DMSO. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 9
<p>Effect of SRE on tartrate-resistant acid phosphatase (TRAP) activity and viability in osteoclastic cells. The “non” sample was incubated in a growth medium with DMSO, and the “cont.” sample was incubated with sRANKL and DMSO. Cell viability was measured by the MTT assay. Data are the mean ± SE of three independent experiments (<span class="html-italic">n</span> = 3).</p> Full article ">
17 pages, 7350 KiB  
Article
Uwhangchungsimwon Inhibits Oxygen Glucose Deprivation/Re-Oxygenation-Induced Cell Death through Neuronal VEGF and IGF-1 Receptor Signaling and Synaptic Remodeling in Cortical Neurons
by Jin Young Hong, Hyunseong Kim, Changhwan Yeo, Wan-Jin Jeon, Junseon Lee, Seung Ho Baek, Yoon Jae Lee and In-Hyuk Ha
Antioxidants 2022, 11(7), 1388; https://doi.org/10.3390/antiox11071388 - 18 Jul 2022
Cited by 1 | Viewed by 1926
Abstract
Uwhangchungsimwon (UCW), a multi-component herbal product, has long been used to treat vascular diseases such as headache, dizziness, high blood pressure, and stroke. Though the prophylactic actions of UCW are well known, insufficient experimental evidence exists on its effectiveness against stroke. Here, we [...] Read more.
Uwhangchungsimwon (UCW), a multi-component herbal product, has long been used to treat vascular diseases such as headache, dizziness, high blood pressure, and stroke. Though the prophylactic actions of UCW are well known, insufficient experimental evidence exists on its effectiveness against stroke. Here, we investigated the mechanism underlying the efficacy of UCW in oxygen glucose deprivation/re-oxygenation (OGD/R)-injury to the primary cortical neurons using an in vitro ischemia model. Neurons secrete vascular endothelial growth factor (VEGF), which acts as a neurotrophic factor in response to an ischemic injury. VEGF modulates neuroprotection and axonal outgrowth by activating the VEGF receptors and plays a critical role in vascular diseases. In this study, cortical neurons were pretreated with UCW (2, 10, and 50 µg/mL) for 48 h, incubated in oxygen-glucose-deprived conditions for 2 h, and further reoxygenated for 24 h. UCW effectively protected neurons from OGD/R-induced degeneration and cell death. Moreover, the role of UCW in sustaining protection against OGD/R injury is associated with activation of VEGF-VEGFR and insulin-like growth factor 1 receptor expression. Therefore, UCW is a potential herbal supplement for the prevention of hypoxic-ischemic neuronal injury as it may occur after stroke. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

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<p>Protective effect of UCW against OGD/R-induced injury in primary cortical neurons. (<b>A</b>) Cell viability of primary cortical neurons pretreated with UCW for 48 h without OGD/R injury and (<b>B</b>) with OGD/R injury using CCK-8 assay (<span class="html-italic">n</span> = 10). (<b>C</b>) Fluorescence intensity of dead cells (red-stained) in neurons pretreated with UCW for 48 h with OGD/R injury (<span class="html-italic">n</span> = 10). (<b>D</b>) Representative images of live and dead assay for live (green) and dead (red) cells in blank, OGD/R injury, and UCW + OGD/R injury. White scale bar = 200 µm. (<b>E</b>) Live-cell imaging of propidium iodide (PI)-stained OGD/R-injured neurons pretreated with UCW at a concentration of 2, 10, and 50 µg/mL for 24 h (<span class="html-italic">n</span> = 6). Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary one-way analysis of variance (ANOVA) with Tukey’s post-hoc analysis as follows: <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01, and <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001 vs. the OGD/R group.</p> Full article ">Figure 2
<p>Antioxidant effect of UCW against OGD/R-induced oxidative injury via inhibition of iNOS-mediated ROS signaling. (<b>A</b>) Representative images of immunocytochemistry for MAP2 (green) and iNOS (red) in blank, OGD/R injury, UCW + OGD/R injury. White scale bar = 50 µm, yellow scale bar = 10 µm. (<b>B</b>) Quantitative analysis of iNOS+/MAP2+ cells estimated based on counts obtained from 10 non-overlapping fields in each group (<span class="html-italic">n</span> = 10). (<b>C</b>) The relative gene expression levels for <span class="html-italic">iNOS</span> mRNA using real-time PCR in each group (<span class="html-italic">n</span> = 4). (<b>D</b>) The percentage of H<sub>2</sub>DCFDA<sup>+</sup> cells in each group by flow cytometry (<span class="html-italic">n</span> = 4). (<b>E</b>) Representative flow cytometry plot images of side scatter (SSC) versus H<sub>2</sub>DCFDA-FITC staining for intracellular ROS levels. Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary ANOVA with Tukey’s post-hoc analysis as follows: <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **** <span class="html-italic">p</span> &lt; 0.0001 vs. OGD/R group.</p> Full article ">Figure 3
<p>Angiogenic potential of UCW for OGD/R-induced hypoxic injury through enhancement of VEGF-VEGFR signaling. (<b>A</b>) Representative images of immunocytochemistry for MAP2 (green) and VEGFR (red) in blank, OGD/R injury, UCW + OGD/R injury. White scale bar = 50 µm, yellow scale bar = 10 µm. (<b>B</b>) Quantitative analysis of the relative VEGFR intensity in each group (<span class="html-italic">n</span> = 8). (<b>C</b>) The relative gene expression levels for <span class="html-italic">VEGFR</span> mRNA using real-time PCR in each group (<span class="html-italic">n</span> = 4). (<b>D</b>) Relative quantification of the VEGFR protein expression using Western blotting in each group (<span class="html-italic">n</span> = 4). Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary ANOVA with Tukey’s post-hoc analysis as follows: <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01 and <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; * <span class="html-italic">p</span> &lt; 0.05 and **** <span class="html-italic">p</span> &lt; 0.0001 vs. the OGD/R group.</p> Full article ">Figure 4
<p>Effect of UCW on neuroprotection against OGD/R-induced injury in primary cortical neurons. (<b>A</b>) Representative images of immunocytochemistry for MAP2 (green) and F-actin (red) in blank, OGD/R injury, UCW + OGD/R injury. White scale bar = 100 µm, red scale bar = 50 µm, and yellow scale bar = 10 µm. (<b>B</b>,<b>C</b>) Quantitative analysis of (<b>B</b>) MAP2 and (<b>C</b>) F-actin intensity in each group (<span class="html-italic">n</span> = 8). (<b>D, E</b>) The relative mRNA expression level for (<b>D</b>) <span class="html-italic">NF200</span> and (<b>E</b>) <span class="html-italic">GAP43</span> using real-time PCR in each group (<span class="html-italic">n</span> = 4). Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary ANOVA with Tukey’s post-hoc analysis as follows: <sup>###</sup> <span class="html-italic">p</span> &lt; 0.001 and <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; *** <span class="html-italic">p</span> &lt; 0.001 and **** <span class="html-italic">p</span> &lt; 0.0001 vs. the OGD/R group.</p> Full article ">Figure 5
<p>Neuroprotective effect of UCW on OGD/R-induced hypoxia damage by activating the IGF-1R signaling in primary cortical neurons. (<b>A</b>) Representative images of immunocytochemistry for IGF-1R (red), MAP2 (green), and GFAP (white) in blank, OGD/R injury, UCW + OGD/R injury. White scale bar = 100 µm, yellow scale bar = 10 µm. (<b>B</b>) Quantitative analysis of IGF-1R intensity estimated based on measurements obtained from 10 non-overlapping fields in each group (<span class="html-italic">n</span> = 10). (<b>C</b>) The relative mRNA expression level for <span class="html-italic">IGF-1R</span> using real-time PCR in each group (<span class="html-italic">n</span> = 4). (<b>D</b>) Relative quantification for the IGF-1R protein expression using Western blotting in each group (<span class="html-italic">n</span> = 4). Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary ANOVA with Tukey’s post-hoc analysis as follows: <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, and **** <span class="html-italic">p</span> &lt; 0.0001 vs. the OGD/R group.</p> Full article ">Figure 6
<p>Preventing synaptic loss of UCW on OGD/R-injured neurons by upregulating synaptic proteins. (<b>A</b>) Representative images of immunocytochemistry for MAP2 (white), SYN1 (green), and PSD95 (red) in blank, OGD/R injury, UCW + OGD/R injury. White scale bar = 100 µm, yellow scale bar = 10 µm. (<b>B</b>,<b>C</b>) Quantitative analysis of (<b>B</b>) SYN1 and (<b>C</b>) PSD95 intensities in each group (<span class="html-italic">n</span> = 10). (<b>D</b>) Quantitative analysis of VEGF<sup>+</sup> or SYN1<sup>+</sup> pixel density in each group (<span class="html-italic">n</span> = 10). (<b>E</b>) Representative images of immunocytochemistry for VEGFR (red) and SYN1 (green) in each group. Yellow scale bar = 10 µm. Data are expressed as the mean ± SEM. Significant differences were analyzed via ordinary ANOVA with Tukey’s post-hoc analysis as follows: <sup>####</sup> <span class="html-italic">p</span> &lt; 0.0001 vs. blank group; **** <span class="html-italic">p</span> &lt; 0.0001 vs. the OGD/R group.</p> Full article ">Scheme 1
<p>Flowchart showing experimental timeline of <span class="html-italic">Uwhangchungsimwon</span> (UCW) treatment and oxygen glucose deprivation/re-oxygenation (OGD/R).</p> Full article ">
14 pages, 820 KiB  
Article
Phytochemical Analysis of the Fruit Pulp Extracts from Annona crassiflora Mart. and Evaluation of Their Antioxidant and Antiproliferative Activities
by Natale Cristine C. Carvalho, Odair S. Monteiro, Claudia Q. da Rocha, Giovanna B. Longato, Robert E. Smith, Joyce Kelly R. da Silva and José Guilherme S. Maia
Foods 2022, 11(14), 2079; https://doi.org/10.3390/foods11142079 - 13 Jul 2022
Cited by 11 | Viewed by 3219
Abstract
Annona crassiflora Mart., the marolo fruit of the Cerrado biome, is one of the most frequently consumed species from the Brazilian Midwest. This study aimed to evaluate the chemical composition and the antioxidant and cytotoxic properties of the fruit pulp of A. crassiflora [...] Read more.
Annona crassiflora Mart., the marolo fruit of the Cerrado biome, is one of the most frequently consumed species from the Brazilian Midwest. This study aimed to evaluate the chemical composition and the antioxidant and cytotoxic properties of the fruit pulp of A. crassiflora collected at Chapada das Mesas, Maranhão, Brazil. The volatile concentrate was identified as mainly ethyl octanoate, ethyl hexanoate, and methyl octanoate. From the ethanol (LFP-E) and ethyl acetate (LFP-A) extracts were identified phenolic acids (p-coumaric, gallic, quinic, and ferulic), flavones and derivatives (apigenin, epicatechin, 2′-5-dimethoxyflavone, 3′,7-dimethoxy-3-hydroxyflavone, kaempferol-3-O-glucoside and 3-O-rutinoside, quercetin-3-O-glucoside, procyanidin B2, and rutin), aporphine alkaloids (xylopine, stephagine, and romucosine), and acetogenin (annonacin). For the LFP-E and LFP-A extracts, the total phenolic compound values were 15.89 and 33.16 mg GAE/g, the flavonoid compound content values were 2.53 and 70.55 mg QE/g, the DPPH radical scavenging activity showed EC50 values of 182.54 and 57.80 µg/mL, and the ABTS radical activity showed TEAC values of 94.66 and 192.61 µM TE/g. The LFP-E extract showed significant cytotoxicity and cell selectivity for the U251-glioma strain, presenting a GI50 value of 21.34 µg/mL, which is close to doxorubicin (11.68 µg/mL), the standard chemotherapeutic drug. The marolo fruit seems to be a promising source for developing innovative and healthy products for the food industry. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Constituents identified in the hydroethanolic and ethyl acetate extracts from fruit pulp of <span class="html-italic">A. crassiflora</span> (see <a href="#foods-11-02079-t001" class="html-table">Table 1</a>).</p> Full article ">Figure 2
<p>Antioxidant capacity of <span class="html-italic">A. crassiflora</span> extracts: (<b>A</b>) EC<sub>50</sub> values of DPPH radical scavenging, (<b>B</b>) Total antioxidant capacity based on ABTS method. <sup>a,b</sup> Different letters represent statistical difference by Tukey test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Growth inhibition of the tumor cell lines after LFP-E extract treatment (48 h).</p> Full article ">
12 pages, 2219 KiB  
Article
Preventative Effects of Antioxidants against PM10 on Serum IgE Concentration, Mast Cell Counts, Inflammatory Cytokines, and Keratinocyte Differentiation Markers in DNCB-Induced Atopic Dermatitis Mouse Model
by Mi Hee Kwack, Jin Seon Bang and Weon Ju Lee
Antioxidants 2022, 11(7), 1334; https://doi.org/10.3390/antiox11071334 - 6 Jul 2022
Cited by 8 | Viewed by 2883
Abstract
Particulate matter (PM) can cause oxidative stress, inflammation, and skin aging. We investigated the effects of antioxidants such as dieckol, punicalagin, epigallocatechin gallate (EGCG), resveratrol, and Siegesbeckiae Herba extract (SHE) against PM < 10 μm (PM10) on serum IgE concentration, mast [...] Read more.
Particulate matter (PM) can cause oxidative stress, inflammation, and skin aging. We investigated the effects of antioxidants such as dieckol, punicalagin, epigallocatechin gallate (EGCG), resveratrol, and Siegesbeckiae Herba extract (SHE) against PM < 10 μm (PM10) on serum IgE concentration, mast cell counts, inflammatory cytokines, and keratinocyte differentiation markers in a 2,4-Dinitrochlorobenzene (DNCB)-induced atopic dermatitis mouse model. Seven-week-old BALB/c mice were sensitized with 2% DNCB. Atopic dermatitis-like lesions were induced on the mice with 0.2% DNCB. Antioxidants and PM10 were applied to the mice for 4 weeks. PM10 increased the serum IgE concentration and spleen weight in mice, and all antioxidants downregulated these parameters. Histological examination showed an increase in epidermal thickness and mast cell counts in response to PM10, and all antioxidants showed a decrease. PM10 upregulates the expression of inflammatory cytokines, including interleukin (IL)-1β, IL-4, IL-6, IL-17α, IL-25, IL-31 and thymic stromal lymphopoietin (TSLP) in mice, and all antioxidants inhibited the upregulation of inflammatory cytokines. ELISA showed the same results as real-time PCR. PM10 downregulates the expression of keratinocyte differentiation markers, including loricrin and filaggrin, in mouse keratinocytes and antioxidants prevented the downregulation of the keratinocyte differentiation markers. Conclusively, PM10 aggravated the DNCB-induced mouse model in serum IgE concentration, mast cell counts, inflammatory cytokine, and keratinocyte differentiation markers. In addition, antioxidants modulated changes in the DNCB-induced mouse model caused by PM10. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Preventative effects of antioxidants against the upregulation of ROS in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated sebocytes and ORS keratinocytes. An increase in the production of reactive oxygen species by PM<sub>10</sub> in cultured (<b>a</b>) sebocytes and (<b>b</b>) ORS keratinocytes was decreased by dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, and Siegesbeckiae Herba extract (SHE). Data are represented as the mean ± SD from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Preventative effects of antioxidants against the upregulation of the serum IgE levels and spleen weights in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated, 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mice. (<b>a</b>) The serum immunoglobulin E (IgE) levels were significantly decreased after the combined treatment with dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, or Siegesbeckiae Herba extract (SHE) compared with those after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± standard deviation (SD) from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05). (<b>b</b>) The spleen weights were also significantly decreased after the combined treatment with dieckol, punicalagin, EGCG, resveratrol, or SHE compared with those after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± SD from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Preventative effects of antioxidants against an increase in epidermal thickness and mast cell counts in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated, 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mice. (<b>a</b>) Antioxidants showed a decrease in epidermal thickness and mast cell counts in the PM<sub>10</sub>-treated, DNCB-induced atopic dermatitis mice. A white arrows indicate epidermal thickness. The data in the bar graphs represent the mean ± standard deviation (SD) from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05). (<b>b</b>) Epidermal thickness was significantly decreased after the combined treatment with dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, or Siegesbeckiae Herba extract (SHE) compared with that after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± SD from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05). (<b>c</b>) The number of mast cells was also significantly decreased after the combined treatment with dieckol, punicalagin, EGCG, resveratrol, or SHE compared with that after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± SD from three independent experiments (* <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Preventative effects of antioxidants on the upregulation of inflammatory cytokines gene expression in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated, 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mice. The gene expressions of interleukin (IL)-1β, IL-4, IL-6, IL-17α, IL-25, IL-31, and thymic stromal lymphopoietin (TSLP) were significantly decreased after combined treatment with dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, or Siegesbeckiae Herba extract (SHE) compared with that after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± standard deviation (SD) of three independent experiments (* <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Preventative effects of antioxidants on the upregulation of inflammatory cytokines protein expression in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated, 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mice. The protein expression of interleukin (IL)-1β, IL-4, IL-17α, and thymic stromal lymphopoietin (TSLP) was also significantly decreased after combined treatment with dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, or Siegesbeckiae Herba extract (SHE) compared with that after PM<sub>10</sub> treatment only. The data in the bar graphs represent the mean ± standard deviation (SD) of three independent experiments (* <span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 6
<p>Preventative effects of antioxidants on the upregulation of the keratinocyte differentiation markers in the particulate matter &lt;10 µm (PM<sub>10</sub>)-treated, 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis mice. The downregulation of keratinocyte differentiation markers by PM<sub>10</sub> alone was prevented by the combination of dieckol, punicalagin, epigallocatechin-3-gallate (EGCG), resveratrol, or Siegesbeckiae Herba extract (SHE).</p> Full article ">
13 pages, 3092 KiB  
Article
Preparation of Mangosteen Peel Extract Microcapsules by Fluidized Bed Spray-Drying for Tableting: Improving the Solubility and Antioxidant Stability
by Sriwidodo Sriwidodo, Reza Pratama, Abd. Kakhar Umar, Anis Yohana Chaerunisa, Afifah Tri Ambarwati and Nasrul Wathoni
Antioxidants 2022, 11(7), 1331; https://doi.org/10.3390/antiox11071331 - 6 Jul 2022
Cited by 10 | Viewed by 4745
Abstract
Mangosteen fruit has been widely consumed and used as a source of antioxidants, either in the form of fresh fruit or processed products. However, mangosteen peel only becomes industrial waste due to its bitter taste, low content solubility, and poor stability. Therefore, this [...] Read more.
Mangosteen fruit has been widely consumed and used as a source of antioxidants, either in the form of fresh fruit or processed products. However, mangosteen peel only becomes industrial waste due to its bitter taste, low content solubility, and poor stability. Therefore, this study aimed to design mangosteen peel extract microcapsules (MPEMs) and tablets to overcome the challenges. The fluidized bed spray-drying method was used to develop MPEM, with hydroxypropyl methylcellulose (HPMC) as the core mixture and polyvinyl alcohol (PVA) as the coating agent. The obtained MPEM was spherical with a hollow surface and had a size of 411.2 µm. The flow rate and compressibility of MPEM increased significantly after granulation. A formula containing 5% w/w polyvinyl pyrrolidone K30 (PVP K30) as a binder had the best tablet characteristics, with a hardness of 87.8 ± 1.398 N, friability of 0.94%, and disintegration time of 25.75 ± 0.676 min. Microencapsulation of mangosteen peel extract maintains the stability of its compound (total phenolic and α-mangosteen) and its antioxidant activity (IC50) during the manufacturing process and a month of storage at IVB zone conditions. According to the findings, the microencapsulation is an effective technique for improving the solubility and antioxidant stability of mangosteen peel extract during manufacture and storage. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Chromatogram of mangosteen peel extract (<b>a</b>) and standard α-mangosteen (<b>b</b>).</p> Full article ">Figure 2
<p>Mangosteen peel extract microcapsules.</p> Full article ">Figure 3
<p>Morphological observation of mangosteen peel extract microcapsules.</p> Full article ">Figure 4
<p>Tablet appearance of formula F1 (<b>a</b>), formula F2 (<b>b</b>), and formula F3 (<b>c</b>).</p> Full article ">
9 pages, 804 KiB  
Article
Maximization of Sulforaphane Content in Broccoli Sprouts by Blanching
by Andrea Mahn, Carmen Elena Pérez, Víctor Zambrano and Herna Barrientos
Foods 2022, 11(13), 1906; https://doi.org/10.3390/foods11131906 - 27 Jun 2022
Cited by 8 | Viewed by 3766
Abstract
Broccoli sprouts are a recognized source of health-promoting compounds, such as glucosinolates, glucoraphanin, and sulforaphane (SFN). Maximization of SFN content can be achieved by technological processing. We investigated the effect of blanching conditions to determine the optimal treatment that maximizes sulforaphane content in [...] Read more.
Broccoli sprouts are a recognized source of health-promoting compounds, such as glucosinolates, glucoraphanin, and sulforaphane (SFN). Maximization of SFN content can be achieved by technological processing. We investigated the effect of blanching conditions to determine the optimal treatment that maximizes sulforaphane content in broccoli sprouts. Broccoli seeds (cv. Traditional) grown under controlled conditions were harvested after 11 days from germination and subjected to different blanching conditions based on a central composite design with temperature and time as experimental factors. Results were analyzed by ANOVA followed by a Tukey test. The optimum conditions were identified through response surface methodology. Blanching increased sulforaphane content compared with untreated sprouts, agreeing with a decrease in total glucosinolates and glucoraphanin content. Temperature significantly affected SFN content. Higher temperatures and shorter immersion times favor glucoraphanin hydrolysis, thus increasing SFN content. The optimum conditions were blanching at 61 °C for 4.8 min, resulting in 54.3 ± 0.20 µmol SFN/g dry weight, representing a 3.3-fold increase with respect to untreated sprouts. This is the highest SFN content reported for sprouts subjected to any treatment so far. The process described in this work may contribute to developing functional foods and nutraceuticals that provide sulforaphane as an active principle. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Standardized effects of the experimental factors and their interactions in blanched broccoli sprouts on (<b>a</b>) sulforaphane, (<b>b</b>) glucoraphanin, and (<b>c</b>) total glucosinolates content. Light blue bars indicate positive effects and pink bars indicate negative effects. The bars exceeding the vertical line had a significant effect on the response.</p> Full article ">Figure 2
<p>Response surfaces obtained for the content of (<b>a</b>) sulforaphane, (<b>b</b>) glucoraphanin, and (<b>c</b>) total glucosinolates.</p> Full article ">
11 pages, 1162 KiB  
Article
Identification of Gene Biomarkers for Tigilanol Tiglate Content in Fontainea picrosperma
by Shahida A Mitu, Praphaporn Stewart, Trong D Tran, Paul W Reddell, Scott F Cummins and Steven M. Ogbourne
Molecules 2022, 27(13), 3980; https://doi.org/10.3390/molecules27133980 - 21 Jun 2022
Cited by 2 | Viewed by 1957
Abstract
Tigilanol tiglate (EBC-46) is a small-molecule natural product under development for the treatment of cancers in humans and companion animals. The drug is currently produced by purification from the Australian rainforest tree Fontainea picrosperma (Euphorbiaceae). As part of a selective-breeding program to increase [...] Read more.
Tigilanol tiglate (EBC-46) is a small-molecule natural product under development for the treatment of cancers in humans and companion animals. The drug is currently produced by purification from the Australian rainforest tree Fontainea picrosperma (Euphorbiaceae). As part of a selective-breeding program to increase EBC-46 yield from F. picrosperma plantations, we investigated potential gene biomarkers associated with biosynthesis of EBC-46. Initially, we identified individual plants that were either high (>0.039%) or low EBC-46 (<0.008%) producers, then assessed their differentially expressed genes within the leaves and roots of these two groups by quantitative RNA sequencing. Compared to low EBC-46 producers, high-EBC-46-producing plants were found to have 145 upregulated genes and 101 downregulated genes in leaves and 53 upregulated genes and 82 downregulated genes in roots. Most of these genes were functionally associated with defence, transport, and biosynthesis. Genes identified as expressed exclusively in either the high or low EBC-46-producing plants were further validated by quantitative PCR, showing that cytochrome P450 94C1 in leaves and early response dehydration 7.1 and 2-alkenal reductase in roots were consistently and significantly upregulated in high-EBC-46 producers. In summary, this study has identified biomarker genes that may be used in the selective breeding of F. picrosperma. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>EBC-46 concentration in leaves of 12 different <span class="html-italic">Fontainea picrosperma</span> plants. (<b>A</b>) HPLC chromatograms at 249nm mAU of (<b>i</b>) EBC-46 analytical standard at 0.1 mg/mL (retention time at 7.496 min), (<b>ii</b>) leaf extract from an example high-EBC-46-producing <span class="html-italic">F. picrosperma</span> plant, and (<b>iii</b>) leaf extract from an example low-EBC46-producing <span class="html-italic">F. picrosperma</span> plant. (<b>B</b>) Average EBC-46 concentration in 12 <span class="html-italic">Fontainea picrosperma</span> plants over one year. Arrowheads indicate plants that were selected for differential gene expression analysis. Error bars represent standard error between monthly analyses. L1-5, low-EBC-46-producing plants; H1-7, high-EBC-46-producing plants. The concentration of EBC-46 in leaves of each high-EBC-46-producing plant (H1-7) was statistically and significantly different than in leaves of each low-EBC-46-producing plant (L1-5).</p> Full article ">Figure 2
<p>Identification of differentially expressed genes in leaf and root tissue from high- and low-EBC-46-producing <span class="html-italic">F. picrosperma</span> plants. (<b>A</b>) Volcano plots showing differentially expressed genes. Adjusted <span class="html-italic">p</span>-value &lt; 0.05. Red colour represents significantly (+2) upregulated and green colour represents significantly (−2) downregulated genes. (<b>B</b>) Venn diagram showing the total number of significantly differentially expressed upregulated (Up) and downregulated (Down) genes in leaf and root tissue, as well as shared genes.</p> Full article ">Figure 3
<p>Validation of differentially expressed genes. Heatmaps showing RNA-seq relative gene expression, log2 fold-change, and <span class="html-italic">p</span>-value for H<sub>EBC-46</sub> and L<sub>EBC-46</sub> <span class="html-italic">F. picrosperma</span> in (<b>A</b>) leaf tissue and (<b>B</b>) root tissue. Four genes were further analysed by RT-qPCR in (<b>C</b>) leaf tissue and (<b>D</b>) root tissue. * represents statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
14 pages, 1373 KiB  
Article
Different Types of Meatballs Enriched with Wild Thyme/Lemon Balm Aqueous Extract—Complex Characterization
by Luiza-Andreea Tănase (Butnariu), Oana-Viorela Nistor, Doina-Georgeta Andronoiu, Gabriel-Dănuț Mocanu, Andreea Veronica Botezatu Dediu and Elisabeta Botez
Molecules 2022, 27(12), 3920; https://doi.org/10.3390/molecules27123920 - 18 Jun 2022
Cited by 3 | Viewed by 1793
Abstract
In the context of the increasing lactation problems among breastfeeding women, the development of a healthy lifestyle is needed. Different variants of pork, turkey, and beef meatballs, with added lemon balm (Melissa officinalis L.) and wild thyme (Thymus serpyllum L.) aqueous [...] Read more.
In the context of the increasing lactation problems among breastfeeding women, the development of a healthy lifestyle is needed. Different variants of pork, turkey, and beef meatballs, with added lemon balm (Melissa officinalis L.) and wild thyme (Thymus serpyllum L.) aqueous extract (6%), were obtained. These herbs were selected and used due to their antioxidant, antimicrobial, and lactogenic potential. Two thermal treatments, hot air convection (180 °C) and steam convection (94 °C), were applied for meatballs processing. The obtained meatballs were further subjected to a complex characterization. The functionality of the plant extracts was proved by the values of total content of polyphenols (2.69 ± 0.02 mg AG/g dw) and flavonoids (3.03 ± 0.24 mg EQ/g dw). FT-IR analysis confirmed the presence of trans-anethole and estragole at 1507–1508 cm−1 and 1635–1638 cm−1, respectively. Costumers’ overall acceptance had a score above 5.5 for all samples, on a scale of 1 to 9. Further analysis and human trials should be considered regarding the use of lactogenic herbs, given their health benefits and availability. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Fourier-transform infrared spectroscopy spectra of (<b>a</b>) standard trans-anethole, (<b>b</b>) standard estragole, (<b>c</b>) meatballs with the addition of aqueous extract of lemon balm/wild thymes; ECPC and ECPA are pork meatballs enriched with wild thyme aqueous extract, processed by hot air and steam convection; ERPC and ERPA are pork meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection; ERCC and ERCA are turkey meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection; ECVC and ECVA are beef meatballs enriched with wild thyme aqueous extract, processed by hot air and steam convection; ERVC and ERVA are beef meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection.</p> Full article ">Figure 2
<p>The in vitro digestion of remanent polyphenolic content in meatball samples in simulated gastric juice (<b>a</b>) and simulated intestinal juice (<b>b</b>); ECPC and ECPA are pork meatballs enriched with wild thyme aqueous extract, processed by hot air and steam convection; ERPC and ERPA are pork meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection; ERCC and ERCA are turkey meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection; ECVC and ECVA are beef meatballs enriched with wild thyme aqueous extract, processed by hot air and steam convection; ERVC and ERVA are beef meatballs enriched with lemon balm aqueous extract, processed by hot air and steam convection.</p> Full article ">Figure 3
<p>Comparative diagram of the sensory attributes specific to meatballs, obtained by (<b>a</b>)—hot air convection and (<b>b</b>)—steam convection; ECPC and ECPA are pork meatballs enriched with wild thyme aqueous extract; ERPC and ERPA are pork meatballs enriched with lemon balm aqueous extract; ERCC and ERCA are turkey meatballs enriched with lemon balm aqueous extract; ECVC and ECVA are beef meatballs enriched with wild thyme aqueous extract; ERVC and ERVA are beef meatballs enriched with lemon balm aqueous extract; HP, HT and HB are pork, turkey, and beef control meatballs, processed by hot air convection. SP, ST, and SB are pork, turkey, and beef control meatballs thermally treated by steam convection.</p> Full article ">Figure 3 Cont.
<p>Comparative diagram of the sensory attributes specific to meatballs, obtained by (<b>a</b>)—hot air convection and (<b>b</b>)—steam convection; ECPC and ECPA are pork meatballs enriched with wild thyme aqueous extract; ERPC and ERPA are pork meatballs enriched with lemon balm aqueous extract; ERCC and ERCA are turkey meatballs enriched with lemon balm aqueous extract; ECVC and ECVA are beef meatballs enriched with wild thyme aqueous extract; ERVC and ERVA are beef meatballs enriched with lemon balm aqueous extract; HP, HT and HB are pork, turkey, and beef control meatballs, processed by hot air convection. SP, ST, and SB are pork, turkey, and beef control meatballs thermally treated by steam convection.</p> Full article ">
10 pages, 1638 KiB  
Article
Potential Application of Gambogic Acid for Retarding Renal Cyst Progression in Polycystic Kidney Disease
by Nutchanard Khunpatee, Kanit Bhukhai, Varanuj Chatsudthipong and Chaowalit Yuajit
Molecules 2022, 27(12), 3837; https://doi.org/10.3390/molecules27123837 - 15 Jun 2022
Cited by 1 | Viewed by 2480
Abstract
Abnormal cell proliferation and accumulation of fluid-filled cysts along the nephrons in polycystic kidney disease (PKD) could lead to a decline in renal function and eventual end-stage renal disease (ESRD). Gambogic acid (GA), a xanthone compound extracted from the brownish resin of the [...] Read more.
Abnormal cell proliferation and accumulation of fluid-filled cysts along the nephrons in polycystic kidney disease (PKD) could lead to a decline in renal function and eventual end-stage renal disease (ESRD). Gambogic acid (GA), a xanthone compound extracted from the brownish resin of the Garcinia hanburyi tree, exhibits various pharmacological properties, including anti-inflammation, antioxidant, anti-proliferation, and anti-cancer activity. However, its effect on inhibiting cell proliferation in PKD is still unknown. This study aimed to determine the pharmacological effects and detailed mechanisms of GA in slowing an in vitro cyst growth model of PKD. The results showed that GA (0.25–2.5 μM) significantly retarded MDCK cyst growth and cyst formation in a dose-dependent manner, without cytotoxicity. Using the BrdU cell proliferation assay, it was found that GA (0.5–2.5 μM) suppressed MDCK and Pkd1 mutant cell proliferation. In addition, GA (0.5–2.5 μM) strongly inhibited phosphorylation of ERK1/2 and S6K expression and upregulated the activation of phosphorylation of AMPK, both in MDCK cells and Pkd1 mutant cells. Taken together, these findings suggested that GA could retard MDCK cyst enlargement, at least in part by inhibiting the cell proliferation pathway. GA could be a natural plant-based drug candidate for ADPKD intervention. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Effect of gambogic acid on MDCK cyst growth and cyst formation. (<b>a</b>) Chemical structure of GA. (<b>b</b>) MDCK cell viability after incubation with DMSO (control) and GA at doses of 0.25, 0.5, 1, 2.5, and 5 μM for 24 h (mean of percent control ± SEM, <span class="html-italic">n</span> = 4, *** <span class="html-italic">p</span> &lt; 0.001, NS; not significant). (<b>c</b>) Inhibitory effect of GA on MDCK cyst growth. The graph shows cyst diameters at day 6, 9, and 12 after treatment with DMSO (control) and GA at doses of 0.25, 0.5, 1, and 2.5 µM from 6 days onwards (four independent experiments, mean ± SEM, <span class="html-italic">n</span> &gt; 65–77 cysts per condition, *** <span class="html-italic">p</span> &lt; 0.001, NS; not significant). (<b>d</b>) Inhibitory effect of GA on MDCK cyst formation. The graph shows the percent of cyst colonies at day 6 after treatment with DMSO (control) and GA at doses of 0.25, 0.5, 1, and 2.5 μM for 6 days (four independent experiments, mean ± SEM, <span class="html-italic">n</span> = 4 wells per condition, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001). (<b>e</b>) A representative light micrograph showing MDCK cyst growth in 3D collagen gels after seeding cells for 6 days, containing 10 µM forskolin and GA at doses of 0, 0.25, 0.5, 1, and 2.5 µM for 6 days. Light micrographs were taken on the indicated days 6, 9, and 12 after cell seeding (scale bar = 100 µm and ×10 magnification).</p> Full article ">Figure 2
<p>Effect of gambogic acid on cell proliferation. MDCK and <span class="html-italic">Pkd1</span> mutant cell proliferation was measured by BrdU incorporation. The graphs show the percentage of cell proliferation in MDCK cells (<b>a</b>) <span class="html-italic">Pkd1<sup>+</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>b</b>) <span class="html-italic">Pkd1<sup>−</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>c</b>) after incubation with DMSO (control), GA at doses of 0.25, 0.5, 1, and 2.5 µM, and 20 µg/mL of blasticidin for 24 h (mean of % control ± SEM, <span class="html-italic">n</span> = 4, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, NS; not significant).</p> Full article ">Figure 3
<p>Dose-dependent effect of gambogic acid on ERK1/2 and S6K protein expression in MDCK and <span class="html-italic">Pkd1</span> mutant cell monolayers. Cells were seeded in a petri-dish (60 × 15 mm) and grown for 24 h. MDCK cells (<b>a</b>) <span class="html-italic">Pkd1<sup>−</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>d</b>), and <span class="html-italic">Pkd1<sup>+</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>g</b>) were incubated with DMSO (control) or GA at doses of 0.25, 0.5, 1, and 2.5 µM for 24 h, and blotted with the indicated antibodies. The representative band intensity of protein expression is shown. Densitometric analysis of p-ERK1/2 and p-S6K expression was normalized to β-actin and is shown as a graph of MDCK cells (<b>b</b>,<b>c</b>), <span class="html-italic">Pkd1<sup>+</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>e</b>,<b>f</b>), and <span class="html-italic">Pkd1<sup>−</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>h</b>,<b>i</b>), respectively (six independent experiments, mean of % control ± SEM, <span class="html-italic">n</span> = 6, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, NS; not significant).</p> Full article ">Figure 4
<p>Effect of gambogic acid on AMPK protein expression in MDCK and <span class="html-italic">Pkd1</span> mutant cell monolayers. Cells were seeded in a petri-dish (60 × 15 mm) and grown for 24 h. MDCK cells (<b>a</b>), <span class="html-italic">Pkd1<sup>−</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>c</b>), and <span class="html-italic">Pkd1<sup>+</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>e</b>) were incubated with DMSO (control), or GA at doses of 0.25, 0.5, 1, and 2.5 µM for 24 h, and were blotted with the indicated antibodies. The representative band intensity of protein expression is shown. Densitometric analysis of p-AMPK/AMPKα expression was normalized to β-actin and is shown as a graph of MDCK cells (<b>b</b>), <span class="html-italic">Pkd1<sup>+</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>d</b>), and <span class="html-italic">Pkd1<sup>−</sup></span><sup>/<span class="html-italic">−</span></sup> cells (<b>f</b>), respectively (six independent experiments, mean of % control ± SEM, <span class="html-italic">n</span> = 6, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, NS; not significant).</p> Full article ">
18 pages, 3815 KiB  
Article
Optimized Ultrasound-Assisted Enzymatic Extraction of Phenolic Compounds from Rosa canina L. Pseudo-Fruits (Rosehip) and Their Biological Activity
by Alexandru Nicolescu, Mihai Babotă, Leilei Zhang, Claudiu I. Bunea, Laura Gavrilaș, Dan C. Vodnar, Andrei Mocan, Gianina Crișan and Gabriele Rocchetti
Antioxidants 2022, 11(6), 1123; https://doi.org/10.3390/antiox11061123 - 6 Jun 2022
Cited by 28 | Viewed by 3718
Abstract
Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used [...] Read more.
Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used for determining the dependence between three variables (i.e., time of extraction, ultrasound amplitude, and the material-to-water ratio) and the total phenolic content of the samples. For EAE, a 2:1:1 pectinase, cellulase, and hemicellulase enzymatic blend was used as pre-treatment for optimized UAE, inducing a higher total phenolic content. The untargeted phenolic profiling approach revealed a great abundance of lower molecular weight phenolics (1.64 mg Eq./g) in UAE-RC extracts, whilst gallic acid (belonging to hydroxybenzoic acid derivatives) was the most abundant individual compound of both extracts. The unsupervised multivariate statistics clearly discriminated the impact of enzymatic pre-treatment on the phenolic profile of RC pseudo-fruits. Finally, Pearson’s correlation coefficients showed that anthocyanins, phenolic acids, and tyrosol derivatives were those compounds mostly correlated to the in vitro antioxidant potential of the extracts, whilst negative and significant (p < 0.05) correlation coefficients were recorded when considering the enzymatic inhibition activities. The highest enzyme-inhibitory activity has been identified against α-glucosidase, which indicates an antidiabetic effect. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The appearance of dried <span class="html-italic">Rosa canina</span> L. pseudo-fruits used for extraction.</p> Full article ">Figure 2
<p>Graphical transposition of the DoE matrix (D-optimal). Each number corresponds to an experimental ID setup, as presented in <a href="#antioxidants-11-01123-t001" class="html-table">Table 1</a>.</p> Full article ">Figure 3
<p>Regression analysis and model interpretation for UAE optimization process; (<b>A</b>) scaled and centered coefficient plot of the process parameter influence; (<b>B</b>) the summary of fit plot for the optimization model.</p> Full article ">Figure 4
<p>Response surface plots for TPC (mg/g) in three cases of SLR (ratio): 1:10, 1:15, and 1:20.</p> Full article ">Figure 5
<p>Total cumulative phenolic content of both ERC and ORC sample extracts. The results are expressed as mg phenolic equivalents (Eq.)/g dry matter (DM).</p> Full article ">Figure 6
<p>Unsupervised multivariate statistics built considering the phenolic profiles of ERC and ORC sample extracts. (<b>A</b>) = heat map based on not averaged hierarchical cluster analysis; (<b>B</b>) = principal component analysis score plot.</p> Full article ">Figure 7
<p>Graphs representing the antioxidant activity (through ABTS and FRAP assays) and the TPC for the obtained freeze-dried extracts. Results expressed as mean ± standard deviations of three parallel measurements.</p> Full article ">Figure 8
<p>Correlogram considering the significant phenolic classes annotated (i.e., anthocyanins, flavones, flavonols, flavan-3-ols, phenolic acids, other phenolics, and stilbenes) and the measured bioactivity (i.e., ABTS and FRAP activity, followed by alpha-glucosidase, tyrosinase, and acetylcholine-esterase inhibitions).</p> Full article ">
18 pages, 4006 KiB  
Article
Optimising the Polyphenolic Content and Antioxidant Activity of Green Rooibos (Aspalathus linearis) Using Beta-Cyclodextrin Assisted Extraction
by Lusani Norah Vhangani, Leonardo Cristian Favre, Guido Rolandelli, Jessy Van Wyk and María del Pilar Buera
Molecules 2022, 27(11), 3556; https://doi.org/10.3390/molecules27113556 - 31 May 2022
Cited by 5 | Viewed by 2231
Abstract
Antioxidant activity associated with green rooibos infusions is attributed to the activity of polyphenols, particularly aspalathin and nothofagin. This study aimed to optimise β-cyclodextrin (β-CD)-assisted extraction of crude green rooibos (CGRE) via total polyphenolic content (TPC) and antioxidant activity assays. Response surface methodology [...] Read more.
Antioxidant activity associated with green rooibos infusions is attributed to the activity of polyphenols, particularly aspalathin and nothofagin. This study aimed to optimise β-cyclodextrin (β-CD)-assisted extraction of crude green rooibos (CGRE) via total polyphenolic content (TPC) and antioxidant activity assays. Response surface methodology (RSM) permitted optimisation of β-CD concentration (0–15 mM), temperature (40–90 °C) and time (15–60 min). Optimal extraction conditions were: 15 mM β-CD: 40 °C: 60 min with a desirability of 0.985 yielding TPC of 398.25 mg GAE·g−1, metal chelation (MTC) of 93%, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging of 1689.7 µmol TE·g−1, ferric reducing antioxidant power (FRAP) of 2097.53 µmol AAE·g−1 and oxygen radical absorbance capacity (ORAC) of 11,162.82 TE·g−1. Aspalathin, hyperoside and orientin were the major flavonoids, with quercetin, luteolin and chrysoeriol detected in trace quantities. Differences (p < 0.05) between aqueous and β-CD assisted CGRE was only observed for aspalathin reporting the highest content of 172.25 mg·g−1 of dry matter for extracts produced at optimal extraction conditions. Positive, strong correlations between TPC and antioxidant assays were observed and exhibited regression coefficient (R2) between 0.929–0.978 at p < 0.001. These results demonstrated the capacity of β-CD in increasing polyphenol content of green rooibos. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Pearson correlation coefficient between total phenolic content and antioxidant activity of crude green rooibos extracts. Total phenolic content versus antioxidant assays (<b>a</b>–<b>d</b>). Antioxidant versus antioxidant (<b>e</b>–<b>j</b>).</p> Full article ">Figure 2
<p>Response surface plots for independent variables for total polyphenolic content (TPC) of crude green rooibos extract: effect of Beta-cyclodextrin and reaction temperature (<b>A</b>) and the effect of Beta-cyclodextrin and reaction time (<b>B</b>).</p> Full article ">Figure 3
<p>HPLC chromatograms of selected flavonoids of CGRE 15 mM: 40 °C: 60 min at 287 nm (<b>A</b>) and 360 nm (<b>B</b>).</p> Full article ">Figure 4
<p>Response surface plots for independent variables for metal chelation (MTC) of crude green rooibos extracts: effect of Beta-cyclodextrin and reaction temperature (<b>A</b>) and the effect of Beta-cyclodextrin and reaction time (<b>B</b>).</p> Full article ">Figure 5
<p>Response surface plots for independent variables for 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging of crude green rooibos extracts: (<b>A</b>) effect of Beta-cyclodextrin and reaction temperature and (<b>B</b>) effect of Beta-cyclodextrin and reaction time.</p> Full article ">Figure 6
<p>Response surface plots for independent variables for ferric reducing antioxidant power (FRAP) of crude green rooibos extracts: (<b>A</b>) effect of Beta-cyclodextrin and reaction temperature and (<b>B</b>) effect of Beta-cyclodextrin and reaction time.</p> Full article ">Figure 7
<p>Response surface plots for independent variables on the oxygen radical absorbance capacity (ORAC) of crude green rooibos extracts. Effect of reaction temperature and Beta-cyclodextrin (<b>A</b>) and effect of reaction temperature and time (<b>B</b>).</p> Full article ">
18 pages, 3152 KiB  
Article
Protective Effect of Resveratrol on Immortalized Duck Intestinal Epithelial Cells Exposed to H2O2
by Ning Zhou, Yong Tian, Hongzhi Wu, Yongqing Cao, Ruiqing Li, Kang Zou, Wenwu Xu and Lizhi Lu
Molecules 2022, 27(11), 3542; https://doi.org/10.3390/molecules27113542 - 31 May 2022
Cited by 10 | Viewed by 2292
Abstract
Resveratrol is a polyphenolic compound with anti-oxidation effects. The mechanisms underlying the antioxidant effects of resveratrol in duck intestinal epithelial cells remain unclear. The protective effects of resveratrol against oxidative stress induced by H2O2 on immortalized duck intestinal epithelial cells [...] Read more.
Resveratrol is a polyphenolic compound with anti-oxidation effects. The mechanisms underlying the antioxidant effects of resveratrol in duck intestinal epithelial cells remain unclear. The protective effects of resveratrol against oxidative stress induced by H2O2 on immortalized duck intestinal epithelial cells (IDECs) were investigated. IDECs were established by transferring the lentivirus-mediated simian virus 40 large T (SV40T) gene into small intestinal epithelial cells derived from duck embryos. IDECs were morphologically indistinguishable from the primary intestinal epithelial cells. The marker protein cytokeratin 18 (CK18) was also detected in the cultured cells. We found that resveratrol significantly increased the cell viability and activity of catalase and decreased the level of intracellular reactive oxygen species and malondialdehyde, as well as the apoptosis rate induced by H2O2 (p < 0.05). Resveratrol up-regulated the expression of NRF2, p-NRF2, p-AKT, and p-P38 proteins and decreased the levels of cleaved caspase-3 and cleaved caspase-9 and the ratio of Bax to Bcl-2 in H2O2-induced IDECs (p < 0.05). Our findings revealed that resveratrol might alleviate oxidative stress by the PI3K/AKT and P38 MAPK signal pathways and inhibit apoptosis by altering the levels of cleaved caspase-3, cleaved caspase-9, Bax, and Bcl-2 in IDECs exposed to H2O2. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Morphology of primary duck intestinal epithelial cells isolated from duck embryos. (<b>A</b>) Cellular morphology in 6 h in culture after isolation. (<b>B</b>) Cellular morphology at passage 3 in culture.</p> Full article ">Figure 2
<p>Characterization of immortalized duck intestinal epithelial cells(IDECs). (<b>A</b>) Morphology of IDECs at passage 30 under microscope. (<b>B</b>) The immunofluorescence study for cytokeratin 18. (<b>C</b>) Primary cells and IDECs were cultured for 7 days and average cell numbers were counted on different days. Data are represented as the mean ± SE (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 3
<p>The IDECs were treated with various concentrations and incubation times of H<sub>2</sub>O<sub>2</sub> (0–500 µM, 2 h and 6 h) and the cell viability was estimated by CCK-8 assay. The data were presented as percent viability of IDECs when treated with different conditions of H<sub>2</sub>O<sub>2</sub>. The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to 0 µM H<sub>2</sub>O<sub>2</sub>.</p> Full article ">Figure 4
<p>The IDECs were treated with 0 µM H<sub>2</sub>O<sub>2</sub>, 200 µM H<sub>2</sub>O<sub>2</sub> and 400 µM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) The ROS production in IDECs treated with different strategies was determined by 2′,7′-dichlorofluorescein diacetate (DCFH-DA). (<b>B</b>) The activities of MDA in IDECs were detected by commercial kits. (<b>C</b>) The activities of CAT in IDECs were detected by commercial kits. (<b>D</b>) The activities of GSH-PX in IDECs were detected by commercial kits. (<b>E</b>) Apoptosis cells were analyzed by flow cytometry using Apoptosis Detection Kit. (<b>F</b>) The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to 0 µM H<sub>2</sub>O<sub>2</sub>.</p> Full article ">Figure 4 Cont.
<p>The IDECs were treated with 0 µM H<sub>2</sub>O<sub>2</sub>, 200 µM H<sub>2</sub>O<sub>2</sub> and 400 µM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) The ROS production in IDECs treated with different strategies was determined by 2′,7′-dichlorofluorescein diacetate (DCFH-DA). (<b>B</b>) The activities of MDA in IDECs were detected by commercial kits. (<b>C</b>) The activities of CAT in IDECs were detected by commercial kits. (<b>D</b>) The activities of GSH-PX in IDECs were detected by commercial kits. (<b>E</b>) Apoptosis cells were analyzed by flow cytometry using Apoptosis Detection Kit. (<b>F</b>) The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to 0 µM H<sub>2</sub>O<sub>2</sub>.</p> Full article ">Figure 5
<p>Effects of resveratrol on the viability of IDECs. (<b>A</b>) IDECs cells were incubated with increasing concentrations of resveratrol for 6 h. * <span class="html-italic">p</span> &lt; 0.05 compared to 0 μM resveratrol. (<b>B</b>) IDECs were pretreated with the resveratrol (5 µM and 10 µM) and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. Then cell viability was measured by CCK-8 assays. The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">Figure 6
<p>IDECs were pretreated with 5 µM resveratrol and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) The ROS production in IDECs treated with different groups was determined by 2′,7′-dichlorofluorescein diacetate (DCFH-DA). (<b>B</b>) The activities of MDA in IDECs were detected by commercial kits. (<b>C</b>) The activities of CAT in IDECs were detected by commercial kits. (<b>D</b>) The activities of GSH-PX in IDECs were detected by commercial kits. (<b>E</b>) Apoptosis cells were analyzed by flow cytometry using Apoptosis Detection Kit. (<b>F</b>) The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">Figure 6 Cont.
<p>IDECs were pretreated with 5 µM resveratrol and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) The ROS production in IDECs treated with different groups was determined by 2′,7′-dichlorofluorescein diacetate (DCFH-DA). (<b>B</b>) The activities of MDA in IDECs were detected by commercial kits. (<b>C</b>) The activities of CAT in IDECs were detected by commercial kits. (<b>D</b>) The activities of GSH-PX in IDECs were detected by commercial kits. (<b>E</b>) Apoptosis cells were analyzed by flow cytometry using Apoptosis Detection Kit. (<b>F</b>) The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">Figure 7
<p>IDECs were pretreated with 5 µM resveratrol for 6 h and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) Protein levels of AKT, p-AKT, P38, p-P38, NRF2, p-NRF2, and GAPDH were detected by Western blot. (<b>B</b>) The expression of AKT, p-AKT, P38, and p-P38 was quantified by densitometry, and data were normalized to GAPDH. (<b>C</b>) The expression of NRF2 and p-NRF2 was quantified, and data were normalized to GAPDH. (<b>D</b>) Histogram of p-AKT/AKT in different treated IDECs. (<b>E</b>) Histogram of p-P38/P38 in different treated IDECs. (<b>F</b>) Histogram of p-NRF2/NRF2 in different treated IDECs. The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to control and <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">Figure 7 Cont.
<p>IDECs were pretreated with 5 µM resveratrol for 6 h and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) Protein levels of AKT, p-AKT, P38, p-P38, NRF2, p-NRF2, and GAPDH were detected by Western blot. (<b>B</b>) The expression of AKT, p-AKT, P38, and p-P38 was quantified by densitometry, and data were normalized to GAPDH. (<b>C</b>) The expression of NRF2 and p-NRF2 was quantified, and data were normalized to GAPDH. (<b>D</b>) Histogram of p-AKT/AKT in different treated IDECs. (<b>E</b>) Histogram of p-P38/P38 in different treated IDECs. (<b>F</b>) Histogram of p-NRF2/NRF2 in different treated IDECs. The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to control and <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">Figure 8
<p>IDECs were pretreated with 5 µM resveratrol for 6 h and then cocultured with 400 μM H<sub>2</sub>O<sub>2</sub> for 6 h. (<b>A</b>) Protein levels of Bax, Bcl-2, caspase-3, cleaved caspase-3, caspase-9, and cleaved caspase-9 were detected by Western blot. (<b>B</b>) The expression of caspase-3, cleaved caspase-3, caspase-9 and cleaved caspase-9 was quantified by densitometry, and data were normalized to GAPDH. (<b>C</b>) Histogram of Bax/Bcl-2 in different treated IDECs. The data were analyzed through one-way ANOVA and are expressed as the mean ± SE (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05 compared to control and <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 compared to H<sub>2</sub>O<sub>2</sub> group.</p> Full article ">
15 pages, 3506 KiB  
Article
Isoquercitrin Attenuates Osteogenic Injury in MC3T3 Osteoblastic Cells and the Zebrafish Model via the Keap1-Nrf2-ARE Pathway
by Xue Li, Dongyue Zhou, Di Yang, Yunhua Fu, Xingyu Tao, Xuan Hu, Yulin Dai and Hao Yue
Molecules 2022, 27(11), 3459; https://doi.org/10.3390/molecules27113459 - 27 May 2022
Cited by 7 | Viewed by 2382
Abstract
Isoquercitrin (IQ) widely exists in natural products, with a variety of pharmacological activities. In this study, the anti-apoptotic and antioxidative activities of IQ were evaluated. IQ showed protective activity against 2, 2′-azobis [2-methylpropionamidine] dihydrochloride (AAPH)-induced cell damage, as well as a marked reduction [...] Read more.
Isoquercitrin (IQ) widely exists in natural products, with a variety of pharmacological activities. In this study, the anti-apoptotic and antioxidative activities of IQ were evaluated. IQ showed protective activity against 2, 2′-azobis [2-methylpropionamidine] dihydrochloride (AAPH)-induced cell damage, as well as a marked reduction in reactive oxygen species (ROS). The evidence of IQ regulating Keap1-Nrf2-ARE and the mitochondrial-mediated Caspase 3 pathway were found in the MC3T3 osteoblastic cell line. Furthermore, IQ significantly decreased ROS production, apoptosis, and lipid peroxidation in AAPH-treated 72 h post-fertilization (hpf) zebrafish, as observed via DCFH-DA, acridine orange (AO), and a 1,3-bis(diphenylphosphino) propane (DPPP) probe, respectively. In AAPH-treated 9 day post-fertilization (dpf) zebrafish, IQ strongly promoted osteogenic development, with increased concentrations by calcein staining, compared with the untreated group. In a molecular docking assay, among all signal proteins, Keap1 showed the strongest affinity with IQ at −8.6 kcal/mol, which might be the reason why IQ regulated the Keap1-Nrf2-ARE pathway in vitro and in vivo. These results indicated that IQ promotes bone development and repairs bone injury, which is valuable for the prevention and treatment of bone diseases. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Effect of IQ on AAPH-induced viability and apoptosis of MC3T3 cells. (<b>a</b>) Cytotoxicity experiment of IQ on MC3T3 cells; (<b>b</b>) concentration screening experiment of AAPH on MC3T3 cells oxidative damage; (<b>c</b>) detection experiment of IQ on the relative level of ROS in MC3T3 cells induced by AAPH; (<b>d</b>) protective experiment of IQ on AAPH-induced MC3T3 cells oxidative damage and apoptosis; (<b>e</b>) staining of apoptotic bodies in the MC3T3 cell nucleus (Hoechst 33342); (<b>f</b>) staining experiment of MC3T3 during nuclear apoptosis (AO/EB). Experiments were performed in triplicate, and the data were expressed as the mean ± SD, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 2
<p>Effect of IQ on AAPH-induced apoptosis cycle of MC3T3 cells. (<b>a</b>) Apoptosis detected by flow cytometry; (<b>b</b>) statistical analysis of cell proportion in different stages of apoptosis. Experiments were performed in triplicate, and the data were expressed as the mean ± SD, ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 3
<p>Effect of IQ on AAPH-induced protein expression of MC3T3 cells. (<b>a</b>) Protein expression levels of the CON group, AAPH group, and IQ-AAPH group on the Keap1-Nrf2-ARE signaling pathway; (<b>b</b>) the protein expression levels of three groups on the Caspase3 signaling pathway. Experiments were performed in triplicate, and the data are expressed as the mean ± SD, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 4
<p>Effect of IQ on AAPH-induced antioxidant and injury repair of zebrafish. (<b>a</b>) ROS staining experiment of 72 hpf zebrafish; (<b>b</b>) apoptotic cell staining experiment of 72 hpf zebrafish; (<b>c</b>) lipid peroxidation staining experiment of 72 hpf zebrafish; (<b>d</b>) heart rate measurement of zebrafish; (<b>e</b>) staining experiment of 9 dpf zebrafish osteogenesis; (<b>f</b>) fluorescence intensity analysis of spine mineralization in 9 dpf zebrafish. Experiments were performed in triplicate, and the data were expressed as the mean ± SD, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 5
<p>3D molecular docking diagram of IQ with Keap1. Affinity = −8.6 kcal/mol. Coordinate parameter: center x = 2.102, center y = 31.44, center z = 68.757, size x = 59.85, size y = 51.3, size z = 44.65; other parameters are the default Auto Dock Tools parameters.</p> Full article ">
16 pages, 4877 KiB  
Article
The Antioxidant and Hypolipidemic Effects of Mesona Chinensis Benth Extracts
by Luhua Xiao, Xiaoying Lu, Huilin Yang, Cuiqing Lin, Le Li, Chen Ni, Yuan Fang, Suifen Mo, Ruoting Zhan and Ping Yan
Molecules 2022, 27(11), 3423; https://doi.org/10.3390/molecules27113423 - 26 May 2022
Cited by 8 | Viewed by 2728
Abstract
In this study, the antioxidant and hypolipidemic effects of Mesona Chinensis Benth (MCB) extracts were evaluated. Seven fractions (F0, F10, F20, F30, F40, F50 and MTF) were obtained from the MCB ethanol extracts. Compared to the commercial antioxidants (vitamin C), MTF and F30 [...] Read more.
In this study, the antioxidant and hypolipidemic effects of Mesona Chinensis Benth (MCB) extracts were evaluated. Seven fractions (F0, F10, F20, F30, F40, F50 and MTF) were obtained from the MCB ethanol extracts. Compared to the commercial antioxidants (vitamin C), MTF and F30 exhibited higher antioxidant activities in the antiradical activity test and the FRAP assay. The half-inhibition concentration (IC50) for MTF and F30 were 5.323 µg/mL and 5.278 µg/mL, respectively. MTF at 200 µg/mL significantly decreased the accumulation of TG in oleic acid (OA)-induced HepG2 cells and reversed the inhibitory effect of Compound C on AMPK (MTF and F30 significantly increased the glucose utilization of insulin-induced HepG2 cells). In addition, the components of MTF were identified by HPLC-MS, which were caffeic acid, quercetin 3-O-galactoside, isoquercetin, astragalin, rosmarinic acid, aromadendrin-3-O-rutinoside, rosmarinic acid-3-O-glucoside and kaempferol-7-O-glucoside. Through statistical correlations by Simca P software, it was found that the main antioxidant and hypolipidemic components of MCB might be caffeic acid, kaempferol-7-O-glucoside, rosmarinic acid-3-O-glucoside and aromadendrin-3-O-rutinoside, which may play important roles in the AMPK pathway. MTF and F30 in MCB could be potential health products for the treatment of hyperlipidemia. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Components of MCB extracts and seven fractions investigated using HPLC. The used wavelength was 320 nm, peaks 1–6 were identified by comparison with authentic standard samples. Peak 1: caffeic acid, Peak 2: quercetin 3-O-galactoside, Peak 3: isoquercetin, Peak 4: astragalin, Peak 5: rosmarinic acid, Peak 6: aromadendrin-3-O-rutinoside, Peak 7: rosmarinic acid-3-O-glucoside, Peak 8: kaempferol-7-O-glucoside.</p> Full article ">Figure 2
<p>Extract ion chromatogram of standards.</p> Full article ">Figure 3
<p>DPPH radical scavenging test results. (<b>A</b>) DPPH free radical scavenging ability of alcohol extracts and aqueous extracts before and after purification. (<b>B</b>) DPPH radical scavenging ability of the fractional eluent. Data are expressed as mean ± standard deviation (<span class="html-italic">n</span> = 5).</p> Full article ">Figure 4
<p>Results of the FRAP method. The in vitro antioxidant activity of MCB is expressed by FRAP values, followed by F30, F10, F40 and F20. EE: alcohol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides. “*” and “**” mean <span class="html-italic">p</span> &lt; 0.01 and <span class="html-italic">p</span> &lt; 0.05, respectively. Data are expressed as mean ± standard deviation (<span class="html-italic">n</span> = 5).</p> Full article ">Figure 5
<p>Effects of MCB extracts on total lipid accumulation in HepG2 cells. (<b>A</b>) the absorbance of MCB extracts on total lipid accumulation. (<b>B</b>) the absorbance of the fractions on total lipid accumulation. NC: control group, Mo: model group, S: drug group, 50: low dose group, M: moderate dose group, H: high dose group, EE: ethanol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides, macroporous resin fractional eluent: F0, F10, F20, F30, F40 and F50. “*” indicates <span class="html-italic">p</span> &lt; 0.05 compared with the NC group. “#” and “##” respectively indicate <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01 compared with the Mo group.</p> Full article ">Figure 6
<p>Effects of MCB extracts on TG level in HepG2 cells. NC: control group, Mo: model group, S: drug group; 50: low dose group, M: moderate dose group, H: high dose group, EE: ethanol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides. “**” indicates <span class="html-italic">p</span> &lt; 0.01 compared with the NC group. “#” and “##” respectively indicate <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01 compared with the Mo group.</p> Full article ">Figure 7
<p>Effects of total flavonoids of MCB on fat accumulation in HepG2 cells (200×). (<b>A</b>) control group, (<b>B</b>) OA group, (<b>C</b>) OA + simvastatin (25 µM) group, (<b>D</b>) OA + MTF (200 µg/mL) group, (<b>E</b>) Compound C (10 µM) group, (<b>F</b>) simvastatin (25 µM) + Compound C (10 µM) group, (<b>G</b>) MTF (200 µg/mL) + Compound C (10 µM) group.</p> Full article ">Figure 8
<p>Effects of total flavonoids (MTF) of MCB on lipid accumulation in HepG2 cells. (<b>A</b>) the absorbance of lipid accumulation was measured by oil red O staining. (<b>B</b>) changes of TG content. “*” indicates <span class="html-italic">p</span> &lt; 0.05 and “**” indicates <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">Figure 9
<p>Effects of MCB extracts on glucose intake in IR-HepG2 cells. NC: control group, Mo: model group, Met: drug group, EE: ethanol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides. “*” and “**” represent <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01 compared with the Mo group, respectively. “#” indicates <span class="html-italic">p</span> &lt; 0.05 compared with the Met group.</p> Full article ">Figure 10
<p>Effects of macroporous resin fractionated eluates on glucose intake in IR-HepG2 cells. NC: control group, Mo: model group, Met: drug group. “*” and “**” represent <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01 compared with the Mo group, respectively. “#” indicates <span class="html-italic">p</span> &lt; 0.05 compared with the Met group.</p> Full article ">Figure 11
<p>Scores of MCB extracts and fractions (PCA). EE: ethanol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides, macroporous resin fractional eluent: F0, F10, F20, F30, F40 and F50.</p> Full article ">Figure 12
<p>Scores of MCB extracts and fractions (OPLS−DA). EE: ethanol extract, AE: aqueous extract, MTF: total flavonoids, MCP: crude polysaccharides, macroporous resin fractional eluent: F0, F10, F20, F30, F40 and F50.</p> Full article ">Figure 13
<p>VIP of MCB extracts and fractions. X1 (Peak 1): caffeic acid, X2 (Peak 2): quercetin 3-O-galactoside, X3 (Peak 3): isoquercetin, X4 (Peak 4): astragalin, X5 (Peak 5): rosmarinic acid, X6 (Peak 6): aromadendrin-3-O-rutinoside, X7 (Peak 7): rosmarinic acid-3-O-glucoside, X8 (Peak 8): kaempferol-7-O-glucoside.</p> Full article ">
15 pages, 2741 KiB  
Article
Glochidion wallichianum Leaf Extract as a Natural Antioxidant in Sausage Model System
by Chantira Wongnen, Naiya Ruzzama, Manat Chaijan, Ling-Zhi Cheong and Worawan Panpipat
Foods 2022, 11(11), 1547; https://doi.org/10.3390/foods11111547 - 25 May 2022
Cited by 7 | Viewed by 2758
Abstract
This study highlighted the role of an 80% ethanolic Mon-Pu (Glochidion wallichianum) leaf extract (MPE), a novel natural antioxidative ingredient, in controlling the oxidative stability and physicochemical properties of a cooked sausage model system (SMS). MPE had a total extractable phenolic [...] Read more.
This study highlighted the role of an 80% ethanolic Mon-Pu (Glochidion wallichianum) leaf extract (MPE), a novel natural antioxidative ingredient, in controlling the oxidative stability and physicochemical properties of a cooked sausage model system (SMS). MPE had a total extractable phenolic content of 16 mg/100 g, with DPPH● scavenging activity, ABTS●+ scavenging activity, and ferric reducing antioxidant power of 2.3, 1.9, and 1.2 mmole Trolox equivalents (TE)/g, respectively. The effects of different concentrations of MPE (0.01–10%, w/w) formulated into SMS on lipid oxidation, protein oxidation, and discoloration were compared to synthetic butylated hydroxyl toluene (BHT; 0.003%, w/w) and a control (without antioxidant). The peroxide value (PV), thiobarbituric acid reactive substances (TBARS), and protein carbonyl contents of SMS tended to increase with increasing MPE concentration (p < 0.05), indicating that high MPE excipient has a pro-oxidative effect. The lowest lipid oxidation (PV and TBARS) and protein carbonyl contents were observed when 0.01% MPE was used to treat SMS (p < 0.05), which was comparable or even greater than BHT-treated SMS. High concentrations (1–10%) of MPE incorporation led to increases in the discoloration of SMS (p < 0.05) with a negligible change in pH of SMS. The water exudate was reduced when MPE was incorporated into SMS compared to control (p < 0.05). Furthermore, MPE at 0.01% significantly reduced lipid oxidation in cooked EMS during refrigerated storage. According to the findings, a low amount of MPE, particularly at 0.01%, in a formulation could potentially maintain the oxidative stability and physicochemical qualities of cooked SMS that are comparable to or better than synthetic BHT. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1

Figure 1
<p>Leaves of <span class="html-italic">Glochidion wallichianum</span> Mull. Arg., commonly known as Mon-Pu (Thai name; MP).</p> Full article ">Figure 2
<p>Total extractable phenolic content, DPPH radical scavenging activity, ABTS radical scavenging activity, and ferric reducing antioxidant power (FRAP) of Mon-Pu leaf extract (MPE). TE = Trolox equivalent. Bars represent the standard deviations from triplicate determinations. Different letters indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Effect of Mon-Pu leaf extract (MPE) on peroxide value (PV) (<b>a</b>), thiobarbituric acid reactive substances (TBARS) (<b>b</b>), and lipid oxidation inhibition (<b>c</b>) of a cooked sausage model system (SMS). Butylated hydroxyl toluene (BHT) at 0.003% was used to compare. The inhibitions of PV and TBARS were calculated relative to the control without MPE and BHT. Bars represent the standard deviations from triplicate determinations. Different letters indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Effect of Mon-Pu leaf extract (MPE) on protein oxidation measured by carbonyl content of a cooked sausage model system (SMS). Butylated hydroxyl toluene (BHT) at 0.003% was used to compare. Bars represent the standard deviations from triplicate determinations. Different letters indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Effect of Mon-Pu leaf extract (MPE) on redness index and percentage discoloration (pattern fill) of a cooked sausage model system (SMS). Butylated hydroxyl toluene (BHT) at 0.003% was used to compare. The percentage discoloration was calculated relative to the control without MEP and BHT. Bars represent the standard deviations from triplicate determinations. Different letters indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 6
<p>Effect of Mon-Pu leaf extract (MPE) on pH (<b>a</b>) and expressible drip (<b>b</b>) of a cooked sausage model system (SMS). Butylated hydroxyl toluene (BHT) at 0.003% was used to compare. Bars represent the standard deviations from triplicate determinations. Different letters indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 7
<p>Changes in peroxide value (PV) (<b>a</b>) and thiobarbituric acid reactive substances (TBARS) (<b>b</b>) of a cooked sausage model system (SMS) added with (☐) and without (■) 0.01% (<span class="html-italic">w</span>/<span class="html-italic">w</span>) Mon-Pu leaf extract (MPE). Bars represent the standard deviations from triplicate determinations. Different lowercase letters in the same treatment and different uppercase letters at the same storage time indicate the significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
15 pages, 2343 KiB  
Article
Protective Effect of Alpinia oxyphylla Fruit against tert-Butyl Hydroperoxide-Induced Toxicity in HepG2 Cells via Nrf2 Activation and Free Radical Scavenging and Its Active Molecules
by Chae Lee Park, Ji Hoon Kim, Je-Seung Jeon, Ju-hee Lee, Kaixuan Zhang, Shuo Guo, Do-hyun Lee, Eun Mei Gao, Rak Ho Son, Young-Mi Kim, Gyu Hwan Park and Chul Young Kim
Antioxidants 2022, 11(5), 1032; https://doi.org/10.3390/antiox11051032 - 23 May 2022
Cited by 8 | Viewed by 2404
Abstract
Alpinia oxyphylla Miq. (Zingiberaceae) extract exerts protective activity against tert-butyl hydroperoxide-induced toxicity in HepG2 cells, and the antioxidant response element (ARE) luciferase activity increased 6-fold at 30 μg/mL in HepG2 cells transiently transfected with ARE-luciferase. To identify active molecules, activity-guided isolation of [...] Read more.
Alpinia oxyphylla Miq. (Zingiberaceae) extract exerts protective activity against tert-butyl hydroperoxide-induced toxicity in HepG2 cells, and the antioxidant response element (ARE) luciferase activity increased 6-fold at 30 μg/mL in HepG2 cells transiently transfected with ARE-luciferase. To identify active molecules, activity-guided isolation of the crude extract led to four sesquiterpenes (1, 2, 5, 6) and two diarylheptanoids (3 and 4) from an n-hexane extract and six sesquiterpenes (7–12) from an ethyl acetate extract. Chemical structures were elucidated by one-dimensional, two-dimensional nuclear magnetic resonance (1D-, 2D-NMR), and mass (MS) spectral data. Among the isolated compounds, eudesma-3,11-dien-2-one (2) promoted the nuclear accumulation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and increased the promoter property of the ARE. Diarylheptanoids, yakuchinone A (3), and 5′-hydroxyl-yakuchinone A (4) showed radical scavenging activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assays. Furthermore, optimization of extraction solvents (ratios of water and ethanol) was performed by comparison of contents of active compounds, ARE-inducing activity, radical scavenging activity, and HepG2 cell protective activity. As a result, 75% ethanol was the best solvent for the extraction of A. oxyphylla fruit. This study demonstrated that A. oxyphylla exerted antioxidant effects via the Nrf2/HO-1 (heme oxygenase-1) pathway and radical scavenging along with active markers eudesma-3,11-dien-2-one (2) and yakuchinone A (3). Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Figure 1
<p>Crude extract of <span class="html-italic">A. oxyphylla</span> induced ARE-luciferase activity in ARE transfected-HepG2 cells. (<b>A</b>) HPLC chromatogram of crude extract of <span class="html-italic">A. oxyphylla</span>. (<b>B</b>) Relative ARE induction activity of crude extract (μg/mL). (<b>C</b>) Relative ARE activity of solvent partitioned extracts from crude extract. (<b>D</b>) HPLC chromatograms of <span class="html-italic">n</span>-hexane extract and EtOAc extract. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control). SUL: sulforaphane was treated as a positive control.</p> Full article ">Figure 2
<p><sup>1</sup>H-NMR spectra of compound <b>4</b> in CDCl<sub>3</sub> and CD<sub>3</sub>OD (400 MHz).</p> Full article ">Figure 3
<p>Key <sup>1</sup>H-<sup>1</sup>H COSY (bold) and HMBC (arrows) correlations for compound <b>4</b>.</p> Full article ">Figure 4
<p>Chemical structures of isolated compounds <b>1</b>–<b>12</b> and their ARE-luciferase induction activity in HepG2 cells. (<b>A</b>) Compounds <b>1</b>–<b>6</b> were isolated from <span class="html-italic">n</span>-hexane extract. (<b>B</b>) Compounds <b>7</b>–<b>12</b> were purified from EtOAc extract. (<b>C</b>) The relative ARE induction activity of each compound to 5 μM sulforaphane (%). Each compound was treated at a concentration of 30 μM. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control). Sulforaphane (SUL) was treated as a positive control.</p> Full article ">Figure 5
<p>Protection against <span class="html-italic">t</span>-BHP-induced toxicity of total extract (<b>A</b>) and pure compounds <b>2</b>–<b>4</b> (<b>B</b>–<b>D</b>) from <span class="html-italic">A</span>. <span class="html-italic">oxyphylla</span>. HepG2-ARE cells were pre-treated with each compound (3, 10, 30, or 100 µM) for 12 h and added <span class="html-italic">t</span>-BHP (300 µM) for 4 h. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control).</p> Full article ">Figure 6
<p>DPPH (<b>A</b>) and ABTS (<b>B</b>) radical scavenging activity of compounds <b>2</b>–<b>4</b>. The treated concentrations of compounds <b>2</b>–<b>4</b> were 3, 10, 30, and 100 μM. Trolox was used as a positive control. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control).</p> Full article ">Figure 7
<p>Induction of the nuclear Nrf2 protein and HO-1 levels in HepG2 cells by compound <b>2</b> or crude extract. Nuclear Nrf2 protein levels in HepG2 cells exposed to compound <b>2</b> for the indicated concentrations (<b>A</b>) and treated with 30 μM compound <b>2</b> for the indicated times (<b>B</b>). Western blot of HO-1 induced by treatment of compound <b>2</b> (<b>C</b>) and crude extract (<b>D</b>) for the indicated concentrations. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control).</p> Full article ">Figure 8
<p>Effects of ethanol-water composition on the DPPH (<b>A</b>), ABTS (<b>B</b>) radical scavenging, ARE-luciferase induction (<b>C</b>), and HepG2 cell protective (<b>D</b>) activity. Data are presented as the mean ± S.D. (<span class="html-italic">n</span> = 3). ** <span class="html-italic">p</span> &lt; 0.01 (compared with the vehicle-treated control).</p> Full article ">Figure 9
<p>Proposed mechanism of active ingredients of <span class="html-italic">A. oxyphylla</span> for the protective activity against <span class="html-italic">t</span>-BHP-induced toxicity in HepG2 cells.</p> Full article ">
20 pages, 3926 KiB  
Review
Updated Pharmacological Effects, Molecular Mechanisms, and Therapeutic Potential of Natural Product Geniposide
by Liping Liu, Qin Wu, Yuping Chen, Guoxiang Gu, Runan Gao, Bo Peng, Yue Wang, Anbang Li, Jipeng Guo, Xinru Xu, Xiaochen Shao, Lingxing Li, Ya Shen and Jihu Sun
Molecules 2022, 27(10), 3319; https://doi.org/10.3390/molecules27103319 - 21 May 2022
Cited by 25 | Viewed by 3061
Abstract
At present, the potential of natural products in new drug development has attracted more and more scientists’ attention, and natural products have become an important source for the treatment of various diseases or important lead compounds. Geniposide, as a novel iridoid glycoside compound, [...] Read more.
At present, the potential of natural products in new drug development has attracted more and more scientists’ attention, and natural products have become an important source for the treatment of various diseases or important lead compounds. Geniposide, as a novel iridoid glycoside compound, is an active natural product isolated from the herb Gardenia jasminoides Ellis (GJ) for the first time; it is also the main active component of GJ. Recent studies have found that geniposide has multiple pharmacological effects and biological activities, including hepatoprotective activity, an anti-osteoporosis effect, an antitumor effect, an anti-diabetic effect, ananti-myocardial dysfunction effect, a neuroprotective effect, and other protective effects. In this study, the latest research progress of the natural product geniposide is systematically described, and the pharmacological effects, pharmacokinetics, and toxicity of geniposide are also summarized and discussed comprehensively. We also emphasize the major pathways modulated by geniposide, offering new insights into the pharmacological effects of geniposide as a promising drug candidate for multiple disorders. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The synthetic method of natural product geniposide. Ethyl 2,3-butadienoate and enone (<span class="html-italic">S</span>)-3b undergo [3+2] cycloaddition reaction under phosphine catalysis to generate cis-fused cyclopenta[c]pyran 4; then, the cis-fused cyclopenta[c]pyran 4 is converted to the natural product iridoid glycoside (+)-geniposide in 10 steps [<a href="#B14-molecules-27-03319" class="html-bibr">14</a>].</p> Full article ">Figure 2
<p>A summary of mechanisms of geniposide on liver injury disease.</p> Full article ">Figure 3
<p>A summary of mechanisms of geniposide on osteoporosis.</p> Full article ">Figure 4
<p>A summary of mechanisms of geniposide on tumors.</p> Full article ">Figure 5
<p>A summary of mechanisms of geniposide on diabetes.</p> Full article ">Figure 6
<p>A summary of mechanisms of geniposide on myocardial dysfunction.</p> Full article ">Figure 7
<p>A summary of mechanisms of geniposide on neurological disease.</p> Full article ">Figure 8
<p>Other therapeutic effects of geniposide.</p> Full article ">
12 pages, 1390 KiB  
Article
Online Extraction–DPPH–HPLC–DAD–QTOF-MS System for Efficient Screening and Identification of Antioxidants from Citrus aurantium L. var. amara (Rutaceae): Integrating Sample Preparation and Antioxidants Profiling
by Yecheng Xiao, Fuhua Fu, Youhe Wei, Shuyun Shi and Yang Shan
Antioxidants 2022, 11(5), 1014; https://doi.org/10.3390/antiox11051014 - 20 May 2022
Cited by 12 | Viewed by 2450
Abstract
The lack of a direct connection between solid edible or medical natural products and bioactive compound profiling is a bottleneck in natural product research and quality control. Here, a novel integrated system, online extraction (OLE)–2,2′-diphenyl-1-picrylhydrazyl (DPPH)–HPLC−DAD−QTOF-MS, was fabricated to extract, screen, and identify [...] Read more.
The lack of a direct connection between solid edible or medical natural products and bioactive compound profiling is a bottleneck in natural product research and quality control. Here, a novel integrated system, online extraction (OLE)–2,2′-diphenyl-1-picrylhydrazyl (DPPH)–HPLC−DAD−QTOF-MS, was fabricated to extract, screen, and identify antioxidants from the whole fruit of Citrus aurantium L. var. amara (CAVA, Rutaceae) simply, rapidly, and efficiently. The system consumes less sample (1.0 mg of CAVA powder) and requires a shorter analytical time (45 min for sample extraction, antioxidants screening, separation, and identification). Eight antioxidant flavonoids were screened and identified, and six available flavanones were sensitively, precisely, and accurately quantified. Two major flavanone glycosides, naringin (50.37 ± 0.43 mg/g) and neohesperidin (38.20 ± 0.27 mg/g), exhibit potent DPPH scavenging activities with IC50 values of 111.9 ± 10.06 and 178.55 ± 11.28 μg/mL. A minor flavanone aglycone, hesperitin (0.73 ± 0.06 mg/g), presents stronger DPPH scavenging activity (IC50, 39.07 ± 2.51 μg/mL). Furthermore, density functional theory calculations demonstrated their electron transport ability and chemical reactivity, which confirmed the screened results. The results indicate that the developed OLE–DPPH–HPLC−DAD−QTOF-MS system provides new perspectives for analysis of antioxidants from complex natural products, which also contribute to the quality evaluation of CAVA. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Full article ">Figure 1
<p>The diagrammatic drawing of OLE–DPPH–HPLC–DAD–QTOF-MS/MS.</p> Full article ">Figure 2
<p>(<b>A</b>) HPLC chromatograms at 254 nm for CAVA (20 μL, 11.5 mg/mL) (a) and 1.0 mg of dried CAVA (b); (<b>B</b>) chromatogram of CAVA-based OLE; (<b>C</b>) OLE–DPPH–HPLC chromatogram at 254 nm for CAVA (1.0 mg); (<b>D</b>) total ion current (TIC) chromatogram for CAVA (1.0 mg) in positive ion mode.</p> Full article ">Figure 3
<p>Structures of potential antioxidants in CAVA.</p> Full article ">Figure 4
<p>TD-DFT calculations of screened flavonoids.</p> Full article ">
16 pages, 2821 KiB  
Article
Antioxidant Activities and Mechanisms of Tomentosin in Human Keratinocytes
by Seyoung Yang, See-Hyoung Park, Sae Woong Oh, Kitae Kwon, Eunbi Yu, Chae Won Lee, Youn Kyoung Son, Changmu Kim, Byoung-Hee Lee, Jae Youl Cho, Youn-Jung Kim and Jongsung Lee
Antioxidants 2022, 11(5), 990; https://doi.org/10.3390/antiox11050990 - 18 May 2022
Cited by 5 | Viewed by 2850
Abstract
Tomentosin, one of natural sesquiterpene lactones sourced from Inula viscosa L., exerts therapeutic effects in various cell types. Here, we investigated the antioxidant activities and the underlying action mechanisms of tomentosin in HaCaT cells (a human keratinocyte cell line). Specifically, we examined the [...] Read more.
Tomentosin, one of natural sesquiterpene lactones sourced from Inula viscosa L., exerts therapeutic effects in various cell types. Here, we investigated the antioxidant activities and the underlying action mechanisms of tomentosin in HaCaT cells (a human keratinocyte cell line). Specifically, we examined the involvement of tomentosin in aryl hydrocarbon receptor (AhR) and nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways. Treatment with tomentosin for up to 60 min triggered the production of reactive oxygen species (ROS), whereas treatment for 4 h or longer decreased ROS production. Tomentosin treatment also induced the nuclear translocation of Nrf2 and upregulated the expression of Nrf2 and its target genes. These data indicate that tomentosin induces ROS production at an early stage which activates the Nrf2 pathway by disrupting the Nrf2–Keap1 complex. However, at a later stage, ROS levels were reduced by tomentosin-induced upregulation of antioxidant genes. In addition, tomentosin induced the phosphorylation of mitogen-activated protein kinases (MAPKs) including p38 MAPK and c-Jun N-terminal kinase (JNK). SB203580 (a p38 MAPK inhibitor) and SP600125 (a JNK inhibitor) attenuated the tomentosin-induced phosphorylation of Nrf2, suggesting that JNK and p38 MAPK signaling pathways can contribute to the tomentosin-induced Nrf2 activation through phosphorylation of Nrf2. Furthermore, N-acetyl-L-cysteine (NAC) treatment blocked both tomentosin-induced production of ROS and the nuclear translocation of Nrf2. These data suggest that tomentosin-induced Nrf2 signaling is mediated both by tomentosin-induced ROS production and the activation of p38 MAPK and JNK. Moreover, tomentosin inhibited the AhR signaling pathway, as evidenced by the suppression of xenobiotic-response element (XRE) reporter activity and the translocation of AhR into nucleus induced by urban pollutants, especially benzo[a]pyrene. These findings suggest that tomentosin can ameliorate skin damage induced by environmental pollutants. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

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Full article ">Figure 1
<p>Tomentosin decreases reactive oxygen species (ROS) in a time- and dose-dependent manner in HaCaT cells. (<b>A</b>) Chemical structure of tomentosin. (<b>B</b>) Cell Counting Kit-8 (CCK-8) was introduced for determining cell viability. HaCaT cells were cultured with the indicated concentrations of tomentosin for 24 h and subjected to CCK-8 analysis. The data are presented as the mean ± SD of triplicates. (<b>C</b>,<b>D</b>) HaCaT cells were pretreated with tomentosin for 24 h prior to treatment with 20 μM 2′,7′ dichlorofluorescin diacetate (DCFDA) for 20 min, and then subjected to fluorescence microscopy (Original magnification = 10×) (<b>C</b>) or microplate-based fluorescence intensity analysis (<b>D</b>). The data are presented as the mean ± SD of triplicates. (<b>E</b>,<b>F</b>) HaCaT cells were pretreated with 10 μM tomentosin for the indicated time periods prior to treatment with 20 μM DCFDA for 20 min and then subjected to fluorescence microscopy (<b>E</b>) (Original magnification = 10×) or microplate-based fluorescence intensity analysis (<b>F</b>). The data are presented as the mean ± SD of triplicates. (<b>G</b>,<b>H</b>) HaCaT cells were pretreated with tomentosin for 24 h prior to treatment with 20 μM DCFDA for 20 min. After 20 min, tert-butyl hydroperoxide (TBHP) was added to a concentration of 55 μM and the cells were incubated for 1 h before being subjected to fluorescence microscopy (<b>G</b>) (Original magnification = 10×) or fluorescence intensity analysis (<b>H</b>). ROS production was assessed using DCFDA. The data are shown as the mean ± SD of triplicates. * <span class="html-italic">p</span> &lt; 0.05 vs. untreated group, ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group, <sup>oo</sup> <span class="html-italic">p</span> &lt; 0.05 vs. TBHP-treated group. TBHP was introduced as a positive control. TBHP, tert-Butyl hydroperoxide.</p> Full article ">Figure 2
<p>Tomentosin exerts antioxidant activity through activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. (<b>A</b>) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of tomentosin. DPPH (0.15 mM) was allowed to react with tomentosin and vitamin C. The data are shown as the mean ± SD of triplicates. (<b>B</b>) HaCaT cells were transfected with an antioxidant response element- (ARE-) luciferase reporter and incubated for 4 h. The cells were then incubated with the indicated concentrations of tomentosin for 24 h and subjected to a luciferase reporter assay. The data are presented as the mean ± SD of triplicates. TBHQ was introduced as a positive control. TBHQ, tert-Butyl hydroquinone. (<b>C</b>–<b>F</b>) HaCaT cells were incubated with 1, 5, or 10 μM tomentosin for 24 h. (<b>C</b>) Protein levels of NAD(P)H-quinone oxidoreductase 1 (NQO1), Nrf2, Kelch-like ECH-associated protein 1 (Keap1), and heme oxygenase-1 (HO-1) were analyzed by Western blot. β-actin was introduced as the control for whole-cell lysates. (<b>D</b>) mRNA levels of HO-1 and NQO1 were analyzed by qRT-PCR. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the control. The results are shown as the mean ± SD of triplicates. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. (<b>E</b>,<b>F</b>) Cells were analyzed by Western blotting (<b>E</b>) of nuclear and cytoplasmic fractions and by immunocytochemistry (<b>F</b>) to assess the level of Nrf2 nuclear translocation. α-tubulin and lamin B1 were introduced as the controls for cytoplasmic and nuclear protein extracts, respectively. NE, nuclear extracts; CE, cytosolic extracts. Scale bar = 50 μm.</p> Full article ">Figure 2 Cont.
<p>Tomentosin exerts antioxidant activity through activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. (<b>A</b>) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of tomentosin. DPPH (0.15 mM) was allowed to react with tomentosin and vitamin C. The data are shown as the mean ± SD of triplicates. (<b>B</b>) HaCaT cells were transfected with an antioxidant response element- (ARE-) luciferase reporter and incubated for 4 h. The cells were then incubated with the indicated concentrations of tomentosin for 24 h and subjected to a luciferase reporter assay. The data are presented as the mean ± SD of triplicates. TBHQ was introduced as a positive control. TBHQ, tert-Butyl hydroquinone. (<b>C</b>–<b>F</b>) HaCaT cells were incubated with 1, 5, or 10 μM tomentosin for 24 h. (<b>C</b>) Protein levels of NAD(P)H-quinone oxidoreductase 1 (NQO1), Nrf2, Kelch-like ECH-associated protein 1 (Keap1), and heme oxygenase-1 (HO-1) were analyzed by Western blot. β-actin was introduced as the control for whole-cell lysates. (<b>D</b>) mRNA levels of HO-1 and NQO1 were analyzed by qRT-PCR. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the control. The results are shown as the mean ± SD of triplicates. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. (<b>E</b>,<b>F</b>) Cells were analyzed by Western blotting (<b>E</b>) of nuclear and cytoplasmic fractions and by immunocytochemistry (<b>F</b>) to assess the level of Nrf2 nuclear translocation. α-tubulin and lamin B1 were introduced as the controls for cytoplasmic and nuclear protein extracts, respectively. NE, nuclear extracts; CE, cytosolic extracts. Scale bar = 50 μm.</p> Full article ">Figure 3
<p>Tomentosin induces the phosphorylation of nuclear factor erythroid 2-related factor 2 (Nrf2) via the p38 and c-Jun <span class="html-italic">N</span>-terminal kinase (JNK) mitogen-activated protein kinase (MAPK) pathway. (<b>A</b>) HaCaT cells were incubated with 1, 5, or 10 μM tomentosin for 24 h. The treated cells were then harvested and analyzed for Nrf2 phosphorylation by Western blotting. β-actin was introduced as the control for whole cell lysates. (<b>B</b>) Cells were treated with 10 μM tomentosin for the indicated time periods (30–90 min). After the treatment period, the cells were collected and analyzed for the phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAPK, and JNK by Western blotting. β-actin was introduced as the control for whole cell lysates. (<b>C</b>) Cells were treated with either 10 μM SB203580 or 10 μM SP600125 in addition to treatment with 10 μM tomentosin for the indicated time periods (30–90 min). After the treatment period, the cells were collected and analyzed for the phosphorylation of JNK and p38 MAPK by Western blotting. β-actin was used as the control for whole-cell lysates. (<b>D</b>,<b>E</b>) Cells were co-treated with 10 μM tomentosin and either (<b>D</b>) 10 μM SB203580 or (<b>E</b>) 10 μM SP600125 for 24 h. After 24 h, the cells were harvested and analyzed for the phosphorylation of Nrf2 by Western blotting. β-actin was introduced as the control for whole-cell lysates.</p> Full article ">Figure 4
<p>Tomentosin-derived ROS production mediates activation of nuclear factor erythroid 2-related factor 2 (Nrf2). (<b>A</b>,<b>B</b>) HaCaT cells were incubated with 10 μM tomentosin for the indicated time periods (5–60 min) and treated with 20 μM 2′,7′ dichlorofluorescin diacetate (DCFDA) for 20 min. The cells were then subjected to fluorescence microscopy (Original magnification = 10×) (<b>A</b>) or fluorescence intensity analysis (<b>B</b>). The results are shown as the mean ± SD of triplicates. * <span class="html-italic">p</span> &lt; 0.05 vs. untreated group. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. (<b>C</b>,<b>D</b>) Cells were treated with 10 μM tomentosin in the presence or absence of 10 mM <span class="html-italic">N</span>-acetyl-<span class="html-italic"><sub>L</sub></span>-cysteine (NAC) for the indicated time points and then treated with 20 μM DCFDA for 20 min. The cells were subjected to fluorescence intensity analysis (<b>C, left</b>) and fluorescence microscopy (<b>C, right</b>)(Original magnification = 10×). The results are shown as the mean ± SD of triplicates. * <span class="html-italic">p</span> &lt; 0.05 vs. untreated group. ## <span class="html-italic">p</span> &lt; 0.01 vs. tomentosin-treated group. (<b>D</b>) Cells were treated with 10 μM tomentosin in the presence of 10 mM NAC for 24 h. The treated cells were collected, and the nuclear and cytoplasmic cellular fractions were analyzed by Western blotting. α-tubulin and lamin B1 were introduced as controls for cytoplasmic and nuclear protein extracts, respectively. NE, nuclear extracts; CE, cytosolic extracts.</p> Full article ">Figure 4 Cont.
<p>Tomentosin-derived ROS production mediates activation of nuclear factor erythroid 2-related factor 2 (Nrf2). (<b>A</b>,<b>B</b>) HaCaT cells were incubated with 10 μM tomentosin for the indicated time periods (5–60 min) and treated with 20 μM 2′,7′ dichlorofluorescin diacetate (DCFDA) for 20 min. The cells were then subjected to fluorescence microscopy (Original magnification = 10×) (<b>A</b>) or fluorescence intensity analysis (<b>B</b>). The results are shown as the mean ± SD of triplicates. * <span class="html-italic">p</span> &lt; 0.05 vs. untreated group. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. (<b>C</b>,<b>D</b>) Cells were treated with 10 μM tomentosin in the presence or absence of 10 mM <span class="html-italic">N</span>-acetyl-<span class="html-italic"><sub>L</sub></span>-cysteine (NAC) for the indicated time points and then treated with 20 μM DCFDA for 20 min. The cells were subjected to fluorescence intensity analysis (<b>C, left</b>) and fluorescence microscopy (<b>C, right</b>)(Original magnification = 10×). The results are shown as the mean ± SD of triplicates. * <span class="html-italic">p</span> &lt; 0.05 vs. untreated group. ## <span class="html-italic">p</span> &lt; 0.01 vs. tomentosin-treated group. (<b>D</b>) Cells were treated with 10 μM tomentosin in the presence of 10 mM NAC for 24 h. The treated cells were collected, and the nuclear and cytoplasmic cellular fractions were analyzed by Western blotting. α-tubulin and lamin B1 were introduced as controls for cytoplasmic and nuclear protein extracts, respectively. NE, nuclear extracts; CE, cytosolic extracts.</p> Full article ">Figure 5
<p>Tomentosin suppresses aryl hydrocarbon receptor (AhR) signaling activated by benzo[a]pyrene (B[a]P). (<b>A</b>,<b>C</b>–<b>H</b>) HaCaT cells were treated with 1, 5, or 10 μM tomentosin in the presence of B[a]P (3 μM) for 24 h. (<b>B</b>) HaCaT cells were incubated with 1, 5, or 10 μM tomentosin in the presence of urban particulate matter (UPM) (100 μg/mL). (<b>A</b>,<b>B</b>) HaCaT cells were transfected with the xenobiotic response element (XRE)-luciferase reporter, and treated with 1, 5, or 10 μM tomentosin in the presence of B[a]P (3 μM) or UPM (100 μg/mL) for 24 h. The cells were then analyzed by a luciferase reporter assay. The results are presented as the mean ± SD of triplicates. (<b>C</b>,<b>D</b>) HaCaT cells were treated with 1, 5, or 10 μM tomentosin in the presence of B[a]P (3 μM) for 24 h. After treatment, the cells were collected and analyzed by Western blotting (<b>C</b>) of the nuclear and cytoplasmic fractions and by immunocytochemical analysis (<b>D</b>) to assess AhR nuclear translocation. α tubulin and lamin B1 were introduced as controls for the cytoplasmic and nuclear protein extracts, respectively. NE, nuclear extracts; CE, cytosolic extracts. Scale bar = 50 μm (<b>E</b>,<b>F</b>) HaCaT cells were treated with 1, 5, or 10 μM tomentosin in the presence of B[a]P (3 μM) for 24 h. (<b>E</b>) CYP1A1 protein expression was analyzed by Western blotting. β-actin was introduced as the control for whole-cell lysates. (<b>F</b>) <span class="html-italic">CYP1A1</span> mRNA levels were analyzed by qRT-PCR; glyceraldeh yde 3-phosphate dehydrogenase (GAPDH) was introduced as the control. The results are presented as the mean ± SD of triplicates. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. ## <span class="html-italic">p</span> &lt; 0.01 vs. B[a]P-treated group.</p> Full article ">Figure 6
<p>Tomentosin reduces the production of intracellular interleukin (IL)-8 and reactive oxygen species (ROS) induced by benzo[a]pyrene (B[a]P). (<b>A</b>,<b>B</b>) HaCaT cells were treated with 1, 5, or 10 μM tomentosin in the presence of B[a]P (3 μM) for 24 h. (<b>A</b>) After treatment, the cells were subjected to enzyme-linked immunosorbent assay (ELISA) for the quantitation of IL-8. The data were confirmed in at least three independent experiments; the results are shown as the mean ± SD of duplicates. (<b>B</b>) After treatment, the cells were incubated with 20 μM dichlorofluorescin diacetate (DCFDA) for 20 min and subjected to fluorescence intensity analysis. The data show the mean ± SD of triplicates. ** <span class="html-italic">p</span> &lt; 0.01 vs. untreated group. # <span class="html-italic">p</span> &lt; 0.05 vs. B[a]P treated group. ## <span class="html-italic">p</span> &lt; 0.01 vs. B[a]P treated group.</p> Full article ">Figure 7
<p>Action mechanism of tomentosin in the AHR/Nrf2-mediated signaling. Tomentosin activates Nrf2 signaling through activation of p38 MAPK and JNK as well as ROS production. Tomentosin also inhibits AHR signaling by suppressing nuclear translocation of AHR. Red line: Action step of tomentosin.</p> Full article ">
16 pages, 3239 KiB  
Article
A Comparative Study on Relieving Exercise-Induced Fatigue by Inhalation of Different Citrus Essential Oils
by Lei Tian, Tan Hu, Shanshan Zhang, Hongyan Zhang, Chenxi Yang, Guiting Chen and Siyi Pan
Molecules 2022, 27(10), 3239; https://doi.org/10.3390/molecules27103239 - 18 May 2022
Cited by 7 | Viewed by 2892
Abstract
Citrus essential oils (CEOs) possess physiological functions due to diverse aroma components. However, evidence for the effects of CEOs on exercise performance and exercise-induced fatigue is limited. The CEOs with discrepancies in components may exert different effects on the amelioration of exercise-induced fatigue. [...] Read more.
Citrus essential oils (CEOs) possess physiological functions due to diverse aroma components. However, evidence for the effects of CEOs on exercise performance and exercise-induced fatigue is limited. The CEOs with discrepancies in components may exert different effects on the amelioration of exercise-induced fatigue. In this study, sweet orange (Citrus sinensis L.) essential oil (SEO), lemon (Citrus limon Osbeck) essential oil (LEO), and bergamot (Citrus bergamia Risso and Poit) essential oil (BEO) were chosen to explore the effect on amelioration of exercise-induced fatigue. Our results demonstrated that SEO and LEO increased the swimming time by 276% and 46.5%, while BEO did not. Moreover, the three CEOs exerted varying effects on mitigating exercise-induced fatigue via inhibiting oxidative stress, protecting muscle injury, and promoting glucose-dependent energy supply. Accordingly, BEO showed the best efficiency. Moreover, the GC-MS and Pearson correlation analysis of BEO showed that the contents of the major components, such as (±)-limonene (32.9%), linalyl butyrate (17.8%), and linalool (7.7%), were significantly positively correlated with relieving exercise-induced fatigue. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Effects of inhalation of the CEOs on weight gain and average food intake in rats. (<b>A</b>) weight gain, (<b>B</b>) average food intake. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 2
<p>Effects of inhalation of the CEOs on swimming exhaustion time in rats. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 3
<p>Effects of the inhalation of the CEOs on energy supply in exercise-induced fatigue rats. (<b>A</b>) Glucose in serum, (<b>B</b>) LG in liver, (<b>C</b>) MG in gastrocnemius. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 4
<p>Effects of the inhalation of the CEOs on metabolite accumulation in exercise-induced fatigue rats. (<b>A</b>) BLA in serum, (<b>B</b>) BUN in serum. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 5
<p>Effects of inhalation of the CEOs on oxidative stress in exercise-induced fatigue rats. (<b>A</b>) SOD in gastrocnemius, (<b>B</b>) GSH-Px in gastrocnemius, (<b>C</b>) MDA in gastrocnemius. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 6
<p>Effects of the inhalation of the CEOs on muscle injury in exercise-induced fatigue rats. (<b>A</b>) CK in serum, (<b>B</b>) LDH in gastrocnemius. Data are the means ± SD. Different lowercase letters indicate significant differences, <span class="html-italic">p</span> &lt; 0.05.</p> Full article ">Figure 7
<p>Pearson correlation analysis between exercise performance, fatigue-associated oxidative stress, energy supply, metabolite accumulation, and muscle injury indices. Blue color indicates a negative correlation, and red color indicates a positive correlation. * indicates a significant correlation between groups (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 8
<p>Experimental procedure for the effects of inhalation of the CEOs on exercise-induced fatigue.</p> Full article ">
15 pages, 2338 KiB  
Article
Hydrolyzed Flavonoids from Cyrtosperma johnstonii with Superior Antioxidant, Antiproliferative, and Anti-Inflammatory Potential for Cancer Prevention
by Ornchuma Naksuriya, Krai Daowtak, Singkome Tima, Siriporn Okonogi, Monika Mueller, Stefan Toegel and Ruttiros Khonkarn
Molecules 2022, 27(10), 3226; https://doi.org/10.3390/molecules27103226 - 18 May 2022
Cited by 3 | Viewed by 1999
Abstract
Cyrtosperma johnstonii is one of the most interesting traditional medicines for cancer treatment. This study aimed to compare and combine the biological activities related to cancer prevention of the flavonoid glycosides rutin (RT) and isorhamnetin-3-o-rutinoside (IRR) and their hydrolysis products quercetin (QT) and [...] Read more.
Cyrtosperma johnstonii is one of the most interesting traditional medicines for cancer treatment. This study aimed to compare and combine the biological activities related to cancer prevention of the flavonoid glycosides rutin (RT) and isorhamnetin-3-o-rutinoside (IRR) and their hydrolysis products quercetin (QT) and isorhamnetin (IR) from C.johnstonii extract. ABTS and MTT assays were used to determine antioxidant activity and cytotoxicity against various cancer cells, as well as normal cells. Anti-inflammatory activities were measured by ELISA. The results showed that the antioxidant activities of the compounds decreased in the order of QT > IR > RT > IRR, while most leukemia cell lines were sensitive to QT and IR with low toxicity towards PBMCs. The reduction of IL-6 and IL-10 secretion by QT and IR was higher than that induced by RT and IRR. The combination of hydrolysis products (QT and IR) possessed a strong synergism in antioxidant, antiproliferative and anti-inflammatory effects, whereas the combination of flavonoid glycosides and their hydrolysis products revealed antagonism. These results suggest that the potential of the combination of hydrolyzed flavonoids from C. johnstonii can be considered as natural compounds for the prevention of cancer. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The hydrolysis scheme of flavonoid glycosides (RT and IRR) to obtain their hydrolysis products (QT and IR).</p> Full article ">Figure 2
<p>Chromatogram of RT (<b>left</b>) and IRR (<b>right</b>) from the extract of <span class="html-italic">C. johnstonii</span>.</p> Full article ">Figure 3
<p>Chromatogram of standard QT (<b>A</b>), QT hydrolyzed from RT using 0.2 M TFA for 30 min (<b>B</b>), QT hydrolyzed from RT using 2.0 M TFA for 30 min (<b>C</b>), standard IR (<b>D</b>), IR hydrolyzed from IRR using 0.2 M TFA for 30 min (<b>E</b>), and IR hydrolyzed from IRR using 2.0 M TFA for 30 min (<b>F</b>).</p> Full article ">Figure 4
<p>Dose-response curves between normal cells (PBMCs) viability and RT (<b>A</b>), QT (<b>B</b>), IRR (<b>C</b>), and IR (<b>D</b>).</p> Full article ">Figure 5
<p>Influence of QT, RT, IR and IRR on the secretion of IL-6 (<b>A</b>) and IL-10 (<b>B</b>) as determined by ELISA assays. Different letters show statistical significance (<span class="html-italic">p</span>-value &lt; 0.05).</p> Full article ">
13 pages, 3220 KiB  
Article
Manipulation of the Phytochemical Profile of Tenderstem® Broccoli Florets by Short Duration, Pre-Harvest LED Lighting
by Faye M. A. Langston, James M. Monaghan, Olivia Cousins, Geoffrey R. Nash, John R. Bows and Gemma Chope
Molecules 2022, 27(10), 3224; https://doi.org/10.3390/molecules27103224 - 18 May 2022
Cited by 2 | Viewed by 2286
Abstract
Light quality has been reported to influence the phytochemical profile of broccoli sprouts/microgreens; however, few studies have researched the influence on mature broccoli. This is the first study to investigate how exposing a mature glasshouse grown vegetable brassica, Tenderstem® broccoli, to different [...] Read more.
Light quality has been reported to influence the phytochemical profile of broccoli sprouts/microgreens; however, few studies have researched the influence on mature broccoli. This is the first study to investigate how exposing a mature glasshouse grown vegetable brassica, Tenderstem® broccoli, to different light wavelengths before harvest influences the phytochemical content. Sixty broccoli plants were grown in a controlled environment glasshouse under ambient light until axial meristems reached >1 cm diameter, whereupon a third were placed under either green/red/far-red LED, blue LED, or remained in the original compartment. Primary and secondary spears were harvested after one and three weeks, respectively. Plant morphology, glucosinolate, carotenoid, tocopherol, and total polyphenol content were determined for each sample. Exposure to green/red/far-red light increased the total polyphenol content by up to 13% and maintained a comparable total glucosinolate content to the control. Blue light increased three of the four indole glucosinolates studied. The effect of light treatments on carotenoid and tocopherol content was inconclusive due to inconsistencies between trials, indicating that they are more sensitive to other environmental factors. These results have shown that by carefully selecting the wavelength, the nutritional content of mature broccoli prior to harvest could be manipulated according to demand. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Total polyphenol content of broccoli samples measured in dry weight across all harvests and light treatments. H in the legend relates to harvest number and T to trial number. Co (control), B (blue), G-R-FR (green/red/far-red). The different letters represent significant differences between samples (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Individual glucosinolate, (<b>a</b>) glucoraphanin, (<b>b</b>) glucoiberin, (<b>c</b>) glucobrassicin, (<b>d</b>) 4-hydroxyglucobrassicin, (<b>e</b>) neoglucobrassicin, (<b>f</b>) 4-methoxyglucobrassicin, and (<b>g</b>) total glucosinolate content of broccoli samples across all the harvests and light treatments. The different letters represent significant differences between samples (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2 Cont.
<p>Individual glucosinolate, (<b>a</b>) glucoraphanin, (<b>b</b>) glucoiberin, (<b>c</b>) glucobrassicin, (<b>d</b>) 4-hydroxyglucobrassicin, (<b>e</b>) neoglucobrassicin, (<b>f</b>) 4-methoxyglucobrassicin, and (<b>g</b>) total glucosinolate content of broccoli samples across all the harvests and light treatments. The different letters represent significant differences between samples (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>(<b>a</b>) Levels of β-carotene, (<b>b</b>) lutein, and (<b>c</b>) γ-tocopherol were measured in broccoli samples across all harvests and light treatments. The different letters represent significant differences between samples (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Images of the plants and lighting conditions. (<b>a</b>) Plants at week 8, trial 1, 19 April 2021, (<b>b</b>) central primary floret, week 12, trial 1, 13 May 2021, (<b>c</b>) LED lighting rack with blue LED lights on the top shelf and four-channel blue, white, red, far-red LED lights on the bottom; (<b>d</b>) LED shelving container when closed.</p> Full article ">
19 pages, 2694 KiB  
Article
Phenotypic Variation and Peel Contribution to Fruit Antioxidant Contents in European and Japanese Plums
by Pavlina Drogoudi and Georgios Pantelidis
Plants 2022, 11(10), 1338; https://doi.org/10.3390/plants11101338 - 18 May 2022
Cited by 11 | Viewed by 2211
Abstract
Herein, we studied the variation in leaf and fruit morphological traits and antioxidant contents in 43 local and foreign cultivars (cvs) grown under the same experimental conditions in the widely cultivated plum species Prunus domestica and Prunus salicina. The peel contribution of [...] Read more.
Herein, we studied the variation in leaf and fruit morphological traits and antioxidant contents in 43 local and foreign cultivars (cvs) grown under the same experimental conditions in the widely cultivated plum species Prunus domestica and Prunus salicina. The peel contribution of fruit bioactive compounds in a serving portion, correlations among the examined parameters, and group patterns in each plum species were also studied. The species and cvs were sufficiently separated. Compared to Japanese cvs, European cvs had less elongated leaves and smaller and sweeter fruit with less total phenol and antioxidant capacities. The Japanese cvs ‘Red ace’ and the widely grown ‘Black Amber’, together with the European ‘Tuleu Dulce’, ‘BlueFre’, and the landrace ‘Asvestochoriou’ make up groups with rich dietary sources of phytochemicals. The peel tissue contained higher total phenols and antioxidant capacities compared to the flesh, while the peel/flesh ratios varied widely among the cvs (6.6-fold). The variation in the antioxidant contents was lower among the cvs calculated per serving portion (3.7-fold); yet the peel tissue contribution was equal to that of the flesh (48.6%), signifying its high nutritive value. We observed increased sweetness in the fruit in the later-harvested cultivars, while cvs with more blue- and red-colored peel generally contained higher antioxidant contents mainly in the European plums. Moreover, larger fruit sizes were positively correlated with larger and more elliptic leaf shapes. In conclusion, the significant role of the genotype and the peel tissue as a source of bioactive compounds in plums were outlined with prospects of utilization in future breeding programs. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Mean (±SE) (<b>a</b>) soluble solid content (SSC, °Brix), titratable acidity (TA, mg citric acid/100 g FW), (<b>b</b>) maturity index (MI = SSC/TA), and ripening date (RD, Julian date) of 43 European and Japanese plums. Colored columns represent local cultivars. Least significant difference; SSC = 1.23, TA = 2.18, SSC/TA = 0.5.</p> Full article ">Figure 2
<p>Mean (±SE) (<b>a</b>) total phenol content (mg gallic acid equivalent/100 g FW), (<b>b</b>) total antioxidant capacity using the DPPH (TAC<sub>DPPH</sub>), and (<b>c</b>) the FRAP radical (TAC<sub>FRAP</sub>) (mg ascorbic acid equivalent/100 g FW), in fruit peel (solid bars) and flesh (rhombus) tissue of 43 European and Japanese plum cvs. Colored columns represent local cultivars. LSD: TPs–peel, 143.5; TPs–flesh, 40.5; TAC<sub>DPPH</sub>–peel, 181.1; TAC<sub>DPPH</sub>–flesh, 38.3; TAC<sub>FRAP</sub>–peel, 172.4; TAC<sub>FRAP</sub>–flesh, 15.9.</p> Full article ">Figure 3
<p>Mean (±SE) (<b>a</b>) total phenols (TPs, mg gallic acid equivalent) and total antioxidant capacity using the (<b>b</b>) DPPH (TAC<sub>DPPH</sub>) and (<b>c</b>) FRAP radicals (TAC<sub>FRAP</sub>) (mg ascorbic acid equivalent), expressed as per serving portion (100 g FW). The percentage (%) contributions of peel and flesh are shown as empty and hatched superimposed columns, respectively. Colored columns represent local cultivars. LSD; total phenols = 35.9, TAC<sub>DPPH</sub> = 33.2, TAC<sub>FRAP</sub> = 19.1.</p> Full article ">Figure 4
<p>(<b>a</b>,<b>c</b>) Segregation and (<b>b</b>,<b>d</b>) factor loadings, of (<b>a</b>,<b>b</b>) 19 European and (<b>c</b>,<b>d</b>) 24 Japanese plum cultivars, on the basis of fruit physical and chemical characters, determined by principal component analysis. Variable annotations are presented in <a href="#plants-11-01338-t003" class="html-table">Table 3</a>.</p> Full article ">Figure 5
<p>Heatmap showing the clustering of fruit phenotyping traits in 19 European plum cultivars using the ClustVis software. The columns correspond to the cultivars and the rows correspond to the fruit phenotypic traits studied. Both the rows and columns were clustered using Euclidean distance and the Ward method.</p> Full article ">Figure 6
<p>Heatmap showing the clustering of fruit phenotyping traits in 24 Japanese plum cultivars using the ClustVis software. The columns correspond to the cultivars and the rows correspond to the fruit phenotypic traits studied. Both rows and columns were clustered using Euclidean distance and the Ward method.</p> Full article ">
23 pages, 3557 KiB  
Article
Integrative Comparative Assessment of Cold Acclimation in Evergreen and Deciduous Iris Species
by Lingmei Shao, Tong Xu, Xiaobin Wang, Runlong Zhang, Xiuyun Wang, Ziming Ren, Jiaping Zhang, Yiping Xia and Danqing Li
Antioxidants 2022, 11(5), 977; https://doi.org/10.3390/antiox11050977 - 16 May 2022
Cited by 3 | Viewed by 2313
Abstract
Cold acclimation (CA) is a strategy which plants have evolved to increase freezing tolerance. Global climate change could obstruct CA and raise the probability of winter injury, especially for evergreens. Hence, understanding the regulatory mechanism of CA is crucial to improve freezing tolerance [...] Read more.
Cold acclimation (CA) is a strategy which plants have evolved to increase freezing tolerance. Global climate change could obstruct CA and raise the probability of winter injury, especially for evergreens. Hence, understanding the regulatory mechanism of CA is crucial to improve freezing tolerance in evergreen plants. A comparative study on a pair of closely related evergreen and deciduous iris species in response to cold through CA was conducive to uncovering and complementing the knowledge of CA. We investigated morphological, physiological and biochemical changes, as well as the expression of associated genes in the functional leaves of both iris species from natural CA to deacclimation. Briefly, fast and strong CA in the evergreen iris might cause early expressions of BAM1, NCED3, GPX6, etc., which leads to strong enzyme activity of starch degradation, abscisic acid biosynthesis and reactive oxygen species scavenging. Additionally, genes belonging to the antioxidant system were mainly induced during deacclimation. These results suggest that interspecies differences in the leaf freezing tolerance of irises are associated with the rate and degree of CA, which activates multiple signaling networks with complex interactions and induces the transcription of cold-responsive genes. Moreover, the ICE–CBF–COR signaling cascade may integrate and initiate diverse cold-responsive pathways during CA of the evergreen iris. The findings of this study provide valuable insight to further research on CA mechanisms and implicate genes which could support breeding strategies in herbaceous perennials under climate changes. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Freezing tolerance and vegetative growth status of leaves in evergreen and deciduous irises under natural cold acclimation and deacclimation. (<b>A</b>) Changes in air temperature from 30 October to 21 March 2021 in the field. The maximum temperature, average temperature and minimum temperature are indicated by red, black and blue lines, respectively. (<b>B</b>) Changes in semi-lethal temperature (LT<sub>50</sub>) for functional leaves in evergreen and deciduous irises. (<b>C</b>) Alterations in the shoot elongation rate in evergreen and deciduous irises. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. The data are the means of three biological replicates, with different letters indicating significant differences between treatments according to Duncan’s multiple range test at <span class="html-italic">p</span> &lt; 0.05. No functional leaves could be sampled from 25 December to 4 March 2021 in deciduous irises. Capital and lowercase letters represent significant differences for relevant parameters within evergreen and deciduous irises (<span class="html-italic">p</span> &lt; 0.05), respectively.</p> Full article ">Figure 2
<p>Leaf microstructures of evergreen and deciduous irises after exposure to low temperatures (during cold acclimation on 27 November 2021). (<b>A</b>) Leaf microstructures of evergreen irises. (<b>B</b>) Leaf microstructures of deciduous irises. Aer, aerenchyma; Cp, chloroplast; E, epidermis; H, hypodermis; SA, stomatal apparatus; V, vascular bundle.</p> Full article ">Figure 3
<p>Physiological and molecular analyses of carbohydrates in the functional leaves of evergreen and deciduous irises under natural cold acclimation and deacclimation. (<b>A–C</b>) Changes in carbohydrate concentrations for functional leaves in evergreen and deciduous irises. For deciduous irises, no functional leaves could be sampled from 25 December to 4 March 2021. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. The data are the means of three biological replicates, with different letters indicating significant differences between treatments according to Duncan’s multiple range test at <span class="html-italic">p</span> &lt; 0.05. The push pin represents standard error. Capital and lowercase letters represent significant differences for relevant parameters within evergreen and deciduous irises (<span class="html-italic">p</span> &lt; 0.05), respectively. (<b>D</b>) Expression levels of sugar-metabolism-related genes expressed in evergreen and deciduous irises. Different colors indicate different levels of gene expression by quantitative real-time PCR; the data in each row were normalized and compared separately; blue indicates downregulation, and red indicates upregulation. Gene names are shown in <a href="#app1-antioxidants-11-00977" class="html-app">Table S2</a>. (<b>E</b>) A schematic illustration of sugar metabolism in evergreen and deciduous irises in this study.</p> Full article ">Figure 4
<p>Measurements of endogenous phytohormones in the functional leaves of evergreen and deciduous irises under natural cold acclimation and deacclimation. Levels of (<b>A</b>) IAA, (<b>B</b>) ABA, (<b>C</b>) JA and (<b>D</b>) GA<sub>3</sub>. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. Values are the means of three biological replicates, and bars represent the standard error. Different letters indicate significant differences according to Duncan’s multiple range test at <span class="html-italic">p</span> &lt; 0.05. Capital and lowercase letters represent significant differences for relevant parameters within evergreen and deciduous irises (<span class="html-italic">p</span> &lt; 0.05), respectively.</p> Full article ">Figure 5
<p>Expression changes of auxin-related genes in functional leaves of evergreen and/or deciduous irises under natural cold acclimation and deacclimation. (<b>A</b>) Expression levels of <span class="html-italic">AMT1</span> for deciduous iris. (<b>B</b>) Expression levels of <span class="html-italic">TIR1</span> for two iris species. (<b>C</b>) Expression levels of <span class="html-italic">GH3.2</span> for evergreen iris. (<b>D</b>) Expression levels of <span class="html-italic">IAA27</span> for two iris species. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. Values are the means of three biological replicates, and bars represent standard error. Different letters indicate significant differences according to Duncan’s multiple range test at <span class="html-italic">p</span> &lt; 0.05. Capital and lowercase letters represent significant differences for relevant parameters within evergreen and deciduous irises (<span class="html-italic">p</span> &lt; 0.05), respectively. Gene names are shown in <a href="#app1-antioxidants-11-00977" class="html-app">Table S2</a>.</p> Full article ">Figure 6
<p>The involvement of plant phytohormone signaling in responses to cold stress and the expression levels of related genes in functional leaves of evergreen and deciduous irises. <span class="html-italic">CBF</span>s can be directly modulated by key components of JA signaling pathways. ABA and GA signaling are thought to facilitate improvements in freezing tolerance in irises, mainly by participating in the ICE–CBF–COR cascade. Arrows indicate promotion, whereas lines ending with a bar suggest suppression. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. Gene names are presented in <a href="#app1-antioxidants-11-00977" class="html-app">Table S2</a>.</p> Full article ">Figure 7
<p>Changes in stress-related indices in the functional leaves of evergreen and deciduous irises and the expression of stress-related genes. Changes in the activities of (<b>A</b>) superoxide dismutase, (<b>B</b>) malondialdehyde, (<b>C</b>) proline and (<b>D</b>) soluble protein in the leaves of both species under natural cold acclimation and deacclimation. Cold acclimation process, sampling from 13 November to 5 February 2021; deacclimation process, sampling from 19 February to 18 March 2021. For the deciduous iris, no functional leaves could be sampled from 25 December to 4 March 2021. The data are the means of three biological replicates, with different letters indicating significant differences between treatments according to Duncan’s multiple range test at <span class="html-italic">p</span> &lt; 0.05. The push pin represents the standard error. Capital and lowercase letters represent significant differences for relevant parameters within evergreen and deciduous irises, respectively (<span class="html-italic">p</span> &lt; 0.05). (<b>E</b>) Expression levels of stress-related genes in evergreen and deciduous irises. Gene names are presented in <a href="#app1-antioxidants-11-00977" class="html-app">Table S2</a>.</p> Full article ">Figure 8
<p>A hypothetical model for iris plants in response to cold stress during winter. Arrows indicate promotion, whereas lines ending with a bar suggest suppression. The blue line indicates that cold signaling partially directly regulates the leaf freezing tolerance of plants through the ICE–CBF–COR cascade. The orange lines indicate phytohormones and sugars that facilitate reactive oxygen species scavenging. The levels of jasmonic acid (JA) were proportional to sucrose concentration in evergreen irises, and the green line indicates a putative pathway. Gene names are shown in <a href="#app1-antioxidants-11-00977" class="html-app">Table S2</a>.</p> Full article ">
25 pages, 7641 KiB  
Article
Structure-Activity Relationship of Pine Nut-Derived Peptides and Their Protective Effect on Nerve-Cell Mitochondria
by Hongyan Lu, Li Fang, Xiyan Wang, Dan Wu, Chunlei Liu, Xiaoting Liu, Ji Wang, Yawen Gao and Weihong Min
Foods 2022, 11(10), 1428; https://doi.org/10.3390/foods11101428 - 15 May 2022
Cited by 5 | Viewed by 2451
Abstract
This study aimed to investigate the structure-activity relationship of the pine nut antioxidant peptide WYPGK and its derivative peptides, and to evaluate the protective effect of the latter on oxidative damage to mitochondrial structure and function in PC12 cells. Molecular docking revealed the [...] Read more.
This study aimed to investigate the structure-activity relationship of the pine nut antioxidant peptide WYPGK and its derivative peptides, and to evaluate the protective effect of the latter on oxidative damage to mitochondrial structure and function in PC12 cells. Molecular docking revealed the derivative peptides WYFGK and WYSGK to have higher affinity to the active region of sirtuin 3 (SIRT3) (−6.08 kcal/mol and −5.87 kcal/mol, respectively), hence indicating that they are promising SIRT3 inducers and antioxidant factors. The derivative peptide WYSGK presented the highest ORAC value (5457.70 µmol TE/g), ABTS scavenging activity (70.05%), and Fe2+-chelating activity (81.70%), followed by WYPGK and WYFGK. Circular dichroism and nuclear magnetic resonance data suggested that the presence of 3-Ser in WYSGK increased its β-sheet content, and that the active hydrogen atoms produced chemical shifts. In H2O2-induced PC12 cells, WYSGK substantially reduced ROS and MDA levels, and increased ATP levels. Transmission electron microscopy and Seahorse Analyze assay proved the peptide WYSGK to significantly alleviate mitochondrial damage and respiratory dysfunction (p < 0.05), thereby implying that a study of structure-activity relationships of the peptides can possibly be an effective approach for the development of functional factors. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The molecular docking mode of (<b>A</b>) WYPGK, (<b>B</b>) WYFGK and (<b>C</b>) WYSGK with SIRT3. The yellow dashed lines stand for hydrogen bonds. The interaction between peptides and the residues of the binding sites in SIRT3 are shown using a 2 D diagram by LigPlus software. (<b>D</b>) WYPGK, (<b>E</b>) WYFGK and (<b>F</b>) WYSGK.</p> Full article ">Figure 1 Cont.
<p>The molecular docking mode of (<b>A</b>) WYPGK, (<b>B</b>) WYFGK and (<b>C</b>) WYSGK with SIRT3. The yellow dashed lines stand for hydrogen bonds. The interaction between peptides and the residues of the binding sites in SIRT3 are shown using a 2 D diagram by LigPlus software. (<b>D</b>) WYPGK, (<b>E</b>) WYFGK and (<b>F</b>) WYSGK.</p> Full article ">Figure 1 Cont.
<p>The molecular docking mode of (<b>A</b>) WYPGK, (<b>B</b>) WYFGK and (<b>C</b>) WYSGK with SIRT3. The yellow dashed lines stand for hydrogen bonds. The interaction between peptides and the residues of the binding sites in SIRT3 are shown using a 2 D diagram by LigPlus software. (<b>D</b>) WYPGK, (<b>E</b>) WYFGK and (<b>F</b>) WYSGK.</p> Full article ">Figure 2
<p>Antioxidant activity assay of WYPGK, WYFGK, and WYSGK. (<b>A</b>) ORAC assay; (<b>B</b>) ABTS radical scavenging activity; (<b>C</b>) Fe<sup>2+</sup> chelating activity. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2 Cont.
<p>Antioxidant activity assay of WYPGK, WYFGK, and WYSGK. (<b>A</b>) ORAC assay; (<b>B</b>) ABTS radical scavenging activity; (<b>C</b>) Fe<sup>2+</sup> chelating activity. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>CD and NMR spectroscopy analysis of WYPGK and WYSGK. (<b>A</b>) CD spectra; (<b>B</b>) secondary structure analysis; the 1H NMR spectroscopy of (<b>C</b>) WYPGK and (<b>D</b>) WYSGK; COSY-NOESY NMR fingerprinting of (<b>E</b>) WYPGK and (<b>F</b>) WYSGK. ** indicates statistically significant differences (<span class="html-italic">p</span> &lt; 0.01).</p> Full article ">Figure 3 Cont.
<p>CD and NMR spectroscopy analysis of WYPGK and WYSGK. (<b>A</b>) CD spectra; (<b>B</b>) secondary structure analysis; the 1H NMR spectroscopy of (<b>C</b>) WYPGK and (<b>D</b>) WYSGK; COSY-NOESY NMR fingerprinting of (<b>E</b>) WYPGK and (<b>F</b>) WYSGK. ** indicates statistically significant differences (<span class="html-italic">p</span> &lt; 0.01).</p> Full article ">Figure 3 Cont.
<p>CD and NMR spectroscopy analysis of WYPGK and WYSGK. (<b>A</b>) CD spectra; (<b>B</b>) secondary structure analysis; the 1H NMR spectroscopy of (<b>C</b>) WYPGK and (<b>D</b>) WYSGK; COSY-NOESY NMR fingerprinting of (<b>E</b>) WYPGK and (<b>F</b>) WYSGK. ** indicates statistically significant differences (<span class="html-italic">p</span> &lt; 0.01).</p> Full article ">Figure 4
<p>Effects of WYPGK and WYSGK on (<b>A</b>) ROS level; (<b>B</b>) MDA level; (<b>C</b>) ATP level; (<b>D</b>) mitochondrial structure and morphology; (<b>E</b>) mitochondrial aspect ratio. Representative quantitative results of mitochondrial length with 10 mitochondria per experiment. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4 Cont.
<p>Effects of WYPGK and WYSGK on (<b>A</b>) ROS level; (<b>B</b>) MDA level; (<b>C</b>) ATP level; (<b>D</b>) mitochondrial structure and morphology; (<b>E</b>) mitochondrial aspect ratio. Representative quantitative results of mitochondrial length with 10 mitochondria per experiment. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4 Cont.
<p>Effects of WYPGK and WYSGK on (<b>A</b>) ROS level; (<b>B</b>) MDA level; (<b>C</b>) ATP level; (<b>D</b>) mitochondrial structure and morphology; (<b>E</b>) mitochondrial aspect ratio. Representative quantitative results of mitochondrial length with 10 mitochondria per experiment. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Effects of WYSGK on the cellular oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Mitochondrial energy metabolism was measured using a Seahorse XF8 extracellular flux analyzer. The mitochondrial (<b>A</b>) OCR curve, (<b>B</b>) basal respiration, (<b>C</b>) ATP production, (<b>D</b>) proton leak, (<b>E</b>) maximal respiration, and (<b>F</b>) spare respiratory capacity. The mitochondrial (<b>G</b>) ECAR curve, (<b>H</b>) glycolysis, (<b>I</b>) glycolysis capacity, and (<b>J</b>) glycolysis reserve. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5 Cont.
<p>Effects of WYSGK on the cellular oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Mitochondrial energy metabolism was measured using a Seahorse XF8 extracellular flux analyzer. The mitochondrial (<b>A</b>) OCR curve, (<b>B</b>) basal respiration, (<b>C</b>) ATP production, (<b>D</b>) proton leak, (<b>E</b>) maximal respiration, and (<b>F</b>) spare respiratory capacity. The mitochondrial (<b>G</b>) ECAR curve, (<b>H</b>) glycolysis, (<b>I</b>) glycolysis capacity, and (<b>J</b>) glycolysis reserve. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5 Cont.
<p>Effects of WYSGK on the cellular oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Mitochondrial energy metabolism was measured using a Seahorse XF8 extracellular flux analyzer. The mitochondrial (<b>A</b>) OCR curve, (<b>B</b>) basal respiration, (<b>C</b>) ATP production, (<b>D</b>) proton leak, (<b>E</b>) maximal respiration, and (<b>F</b>) spare respiratory capacity. The mitochondrial (<b>G</b>) ECAR curve, (<b>H</b>) glycolysis, (<b>I</b>) glycolysis capacity, and (<b>J</b>) glycolysis reserve. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5 Cont.
<p>Effects of WYSGK on the cellular oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Mitochondrial energy metabolism was measured using a Seahorse XF8 extracellular flux analyzer. The mitochondrial (<b>A</b>) OCR curve, (<b>B</b>) basal respiration, (<b>C</b>) ATP production, (<b>D</b>) proton leak, (<b>E</b>) maximal respiration, and (<b>F</b>) spare respiratory capacity. The mitochondrial (<b>G</b>) ECAR curve, (<b>H</b>) glycolysis, (<b>I</b>) glycolysis capacity, and (<b>J</b>) glycolysis reserve. Data are presented as means ± SD from three replicates. Different letters denote statistically significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
12 pages, 289 KiB  
Article
Stevia (Stevia rebaudiana) Improves Carotenoid Content in Eggs When Fed to Laying Hens
by Vasil Radoslavov Pirgozliev, Isobel Margaret Whiting, Kristina Kljak, Stephen Charles Mansbridge, Atanas Georgiev Atanasov, Stephen Paul Rose and Stanimir Bojidarov Enchev
Foods 2022, 11(10), 1418; https://doi.org/10.3390/foods11101418 - 13 May 2022
Cited by 13 | Viewed by 3356
Abstract
Stevia rebaudiana Bertoni is a shrub with leaves that have a high concentration of carotenoids such as lutein and zeaxanthin. Egg yolks are a bioavailable source of lutein and zeaxanthin. The consumption of these carotenoids has been linked with improved human health. To [...] Read more.
Stevia rebaudiana Bertoni is a shrub with leaves that have a high concentration of carotenoids such as lutein and zeaxanthin. Egg yolks are a bioavailable source of lutein and zeaxanthin. The consumption of these carotenoids has been linked with improved human health. To investigate the impact of dried stevia leaves at 0%, 1% and 2% on the quality variables, the chemical composition and antioxidant content of eggs, the experiment involved 90 Hy-Line Brown laying hens, housed in 30 enriched layer cages, in groups of three from 22 to 26 weeks of age. The impact on the internal qualities of stored eggs was also examined. Yolks from hens fed stevia had an enriched color compared with the controls. At the end of the experiment, the whole egg, without shell, of birds fed 2% stevia had a higher total carotenoid content (p < 0.001) compared with birds fed 1% and 0% stevia, i.e., 5.16 (µg/g), 4.23 (µg/g) and 2.96 (µg/g), respectively. Storage reduced albumen height and increased albumen pH (p < 0.001). Stevia supplementation did not interact (p > 0.05) with storage time among the egg quality variables. Consuming eggs from hens fed stevia may increase carotenoids in human diet. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
12 pages, 715 KiB  
Article
Impact of Leaf Removal on Phenolics and Antioxidant Activity of Trebbiano Berries (Vitis vinifera L.)
by Mike Frank Quartacci, Cristina Sgherri and Calogero Pinzino
Plants 2022, 11(10), 1303; https://doi.org/10.3390/plants11101303 - 13 May 2022
Cited by 1 | Viewed by 1767
Abstract
Leaf removal is a canopy management practice widely applied in viticulture to enhance the phenol composition and concentration of grapes, which then results in improved wine quality. Many studies were carried out on red berried varieties, but information on white ones is scanty. [...] Read more.
Leaf removal is a canopy management practice widely applied in viticulture to enhance the phenol composition and concentration of grapes, which then results in improved wine quality. Many studies were carried out on red berried varieties, but information on white ones is scanty. The aim of the study was to assess the effect of basal leaf defoliation in post fruit set on the phenol composition, ascorbate level and antioxidant activity of Trebbiano grapes. Electron paramagnetic resonance was also employed to monitor the decay kinetics of 1,1-diphenyl-2-picrylhydrazyl which allowed the identification of antioxidants with different action rates. The results show that defoliation caused an increase in the phenolic acid (hydroxycinnamic and hydroxybenzoic acids) and flavonol concentrations of berries without changes in the composition. Both ascorbate and antioxidant activity were also enhanced in the berries from defoliated vines. Besides increasing the number of fast-rate antioxidants, leaf removal resulted in the formation of intermediate-rate ones. In the Trebbiano variety, leaf removal in the post fruit set may represent an effective strategy to enhance the phenolic composition and the antioxidant defense system of berries. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Total phenols (<b>A</b>), antioxidant activity (<b>B</b>) and ascorbate (<b>C</b>) in grape berries from control and defoliated vines of <span class="html-italic">Vitis vinifera</span> (Trebbiano variety). C, control; D, defoliated; AsA, reduced ascorbate. Data are reported as mean values ± standard error. For each compound means accompanied by different letters are significantly different at <span class="html-italic">p</span> ≤ 0.05.</p> Full article ">Figure 2
<p>EPR decay kinetics of 1,1-diphenyl-2-picrylhydrazyl (DPPH•) obtained from berry extracts from control and defoliated vines of <span class="html-italic">Vitis vinifera</span> (Trebbiano variety). C, control; D, defoliated.</p> Full article ">
12 pages, 657 KiB  
Article
Exploring Lignans, a Class of Health Promoting Compounds, in a Variety of Edible Oils from Brazil
by Roberta Tardugno, Nicola Cicero, Rosaria Costa, Vincenzo Nava and Rossella Vadalà
Foods 2022, 11(10), 1386; https://doi.org/10.3390/foods11101386 - 11 May 2022
Cited by 5 | Viewed by 2224
Abstract
Lignans, a group of polyphenols, have been identified in eight cold pressed oils from fruits, nuts, and seeds, retrieved from the Brazilian market. The oils under investigation were avocado, Brazilian nut, canola, coconut, grapeseed, macadamia, palm, and pequi. Olive oil was selected as [...] Read more.
Lignans, a group of polyphenols, have been identified in eight cold pressed oils from fruits, nuts, and seeds, retrieved from the Brazilian market. The oils under investigation were avocado, Brazilian nut, canola, coconut, grapeseed, macadamia, palm, and pequi. Olive oil was selected as a reference oil, since numerous data on its lignan content are available in literature. The qualitative and quantitative profiles were obtained, after extraction, by means of UFLC-ESI-MS/MS analyses. The total lignan content showed a high variability, ranging from 0.69 mg·Kg−1 (pequi) to 7.12 mg·Kg−1 (grapeseed), with the highest content registered for olive oil. Seven lignans were quantified, matairesinol and pinoresinol being the most abundant. The LC-MS/MS method was validated, showing linearity in the range of 12.5–212.5 mg·Kg−1, LOD in the range of 0.18–11.37 mg·Kg−1, and LOQ in the range of 0.53–34.45 mg·Kg−1. Additionally, part of the study was focused on the evaluation of the flavor profile, this being a key element in consumers’ evaluations, by means of HS-SPME-GC. In total, 150 volatile compounds were determined in the eight oils, with identified fractions ranging from 91.85% (avocado) to 96.31% (canola), with an average value of 94.1%. Groups of components contributed characteristically to the flavour of each oil. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>HS-SPME-GC-FID chromatograms of the eight Brazilian cold-pressed oils.</p> Full article ">
17 pages, 318 KiB  
Article
Chemical Composition, Antioxidant and Anti-Enzymatic Activity of Golden Root (Rhodiola rosea L.) Commercial Samples
by Milena Polumackanycz, Pawel Konieczynski, Ilkay Erdogan Orhan, Nurten Abaci and Agnieszka Viapiana
Antioxidants 2022, 11(5), 919; https://doi.org/10.3390/antiox11050919 - 7 May 2022
Cited by 12 | Viewed by 3093
Abstract
The aim of the study was to compare the chemical composition of the water and hydromethanolic extracts of R. rosea commercial samples in relation to their biological activity. For this purpose, the HPLC method was used for the determination of eleven phenolic compounds [...] Read more.
The aim of the study was to compare the chemical composition of the water and hydromethanolic extracts of R. rosea commercial samples in relation to their biological activity. For this purpose, the HPLC method was used for the determination of eleven phenolic compounds and AAS/AES was used for determination of five essential elements. Moreover, the contents of total phenolic, total flavonoid, total phenolic acids, and L(+)-ascorbic acid were determined. The antioxidant activity was assessed by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS radical scavenging activity, ferric-reducing/antioxidant power (FRAP), and cupric-reducing antioxidant capacity (CUPRAC) assays, while the inhibitory activity against AChE and BChE enzymes was determined using Ellman’s method. The results showed that the hydromethanolic extracts of R. rosea were richer in phenolic compounds and showed higher antioxidant and neurobiological activity than the water extracts. However, the water extracts gave higher contents of determined elements. Among the individual phenolic compounds gallic acid (2.33 mg/g DW) and sinapic acid (386.44 µg/g DW) had the highest concentrations in the hydromethanolic and water extracts, respectively. Moreover, the most extracts were observed to be more efficient on BChE. Moreover, the correlation analysis indicated a high positive relationship between chemical composition and biological activity in both extracts of R. rosea. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
16 pages, 5924 KiB  
Article
Isolation, Structural Elucidation, Antioxidant and Hypoglycemic Activity of Polysaccharides of Brassica rapa L.
by Wenyang Cao, Chenxi Wang, Xiayidan Mayhesumu, Le Pan, Yan Dang, Abulimiti Yili, Aytursun Abuduwaili and Sanawar Mansur
Molecules 2022, 27(9), 3002; https://doi.org/10.3390/molecules27093002 - 7 May 2022
Cited by 10 | Viewed by 2455
Abstract
The aim of this study was to investigate the effects of microwave ultrasonic-assisted extraction (MUAE) on the content, structure, and biological functions of Brassica rapa L. polysaccharide (BRP). Response surface methodology (RSM) was used to optimize the parameters of MUAE, and it obtained [...] Read more.
The aim of this study was to investigate the effects of microwave ultrasonic-assisted extraction (MUAE) on the content, structure, and biological functions of Brassica rapa L. polysaccharide (BRP). Response surface methodology (RSM) was used to optimize the parameters of MUAE, and it obtained a polysaccharide with yield of 21.802%. Then, a neutral polysaccharide named BRP-1-1 with a molecular weight of 31.378 kDa was isolated and purified from BRP using DEAE-650 M and Sephadex G-100. The structures of the BRP-1-1 were elucidated through a combination of FT-IR, GC-MS, NMR, and methylation analysis. The results showed that BRP-1 consisted of mannose (Man) and glucose (Glu) in a molar ratio of 7.62:1. The backbone of BRP-1-1 mainly consisted of →6)-α-D-Glup-(1→4-β-D-Glup-(1→2)-α-D-Manp-(1→2)-α-D-Glup-(1→, the branch was [T-α-D-Manp-(1]n→. BRP-1-1 intervention significantly inhibited α-glucosidase activity; an inhibition rate of 44.623% was achieved at a concentration of 0.5 mg/mL. The results of the in vitro biological activity showed that BRP-1-1 has good antioxidant and hypoglycemic activity, suggesting that BRP-1-1 could be developed as a functional medicine. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Effect of the single factor on the BRP yield. (<b>a</b>) Ratio of water to material (mL/g); (<b>b</b>) extraction temperature (°C); (<b>c</b>) microwave-ultrasound time (min) and (<b>d</b>) ultrasound power (w).</p> Full article ">Figure 1 Cont.
<p>Effect of the single factor on the BRP yield. (<b>a</b>) Ratio of water to material (mL/g); (<b>b</b>) extraction temperature (°C); (<b>c</b>) microwave-ultrasound time (min) and (<b>d</b>) ultrasound power (w).</p> Full article ">Figure 2
<p>The response surface (3D) shows the effect of each variable (<b>a</b>–<b>f</b>) on the BRP content.</p> Full article ">Figure 3
<p>(<b>a</b>) Chromatogram of BRP on DEAE-650 column, (<b>b</b>) chromatogram of BRP-1 on Sephadex G-100 column; (<b>c</b>) the Mw distribution of BRP-1-1; (<b>d</b>) the FT-IR spectra of RPP-1-1.</p> Full article ">Figure 4
<p><sup>1</sup>H NMR (<b>a</b>), <sup>13</sup>C NMR (<b>b</b>), NOESY (<b>c</b>), HSQC (<b>d</b>), COSY (<b>e</b>), and HMBC spectra of BRP-1-1 (<b>f</b>); the predicted structure of BRP-1-1 (<b>g</b>).</p> Full article ">Figure 4 Cont.
<p><sup>1</sup>H NMR (<b>a</b>), <sup>13</sup>C NMR (<b>b</b>), NOESY (<b>c</b>), HSQC (<b>d</b>), COSY (<b>e</b>), and HMBC spectra of BRP-1-1 (<b>f</b>); the predicted structure of BRP-1-1 (<b>g</b>).</p> Full article ">Figure 5
<p>(<b>a</b>) SEM of BRP-1-1 observed at 20 KX; (<b>b</b>) XRD characterization of BRP-1-1; (<b>c</b>) variation curves of λ<sub>max</sub> of Congo red + BRP-1-1 and Congo red solution.</p> Full article ">Figure 6
<p>(<b>a</b>) DPPH<sup>+</sup> scavenging rate; (<b>b</b>) hydroxyl scavenging rate; (<b>c</b>) ABTS<sup>+</sup> scavenging rate.</p> Full article ">Figure 7
<p>(<b>a</b>) α-Glucosidase inhibitory assay; (<b>b</b>) α-amylase inhibitory assay.</p> Full article ">
10 pages, 1564 KiB  
Article
Content Variations in Oleocanthalic Acid and Other Phenolic Compounds in Extra-Virgin Olive Oil during Storage
by Jasmine Esposito Salsano, Maria Digiacomo, Doretta Cuffaro, Simone Bertini and Marco Macchia
Foods 2022, 11(9), 1354; https://doi.org/10.3390/foods11091354 - 6 May 2022
Cited by 11 | Viewed by 2645
Abstract
The health benefits of extra-virgin olive oil (EVOO) are strictly linked to the presence of phenolic compounds, which exhibit numerous nutraceutical properties. In EVOO, the most important class of phenolic compounds is represented by secoiridoids (oleacein and oleocanthal). EVOO is constantly subjected to [...] Read more.
The health benefits of extra-virgin olive oil (EVOO) are strictly linked to the presence of phenolic compounds, which exhibit numerous nutraceutical properties. In EVOO, the most important class of phenolic compounds is represented by secoiridoids (oleacein and oleocanthal). EVOO is constantly subjected to degradation processes, including hydrolytic and oxidative reactions that influence its phenolic composition. In particular, the hydrolytic reactions determine the transformation of oleocanthal and oleacein into the corresponding phenyl-alcohols, tyrosol, and hydroxytyrosol. Furthermore, oleocanthal by oxidation processes can be converted to oleocanthalic acid. In this study, we evaluated the phenolic composition of three EVOO samples kept at different storage conditions for 15 months, focusing on the variation of oleocanthalic acid content. Specifically, the samples were stored at 4 °C in darkness and at 25 °C with light exposure. The results of our analyses highlighted that in EVOOs exposed to light and maintained at 25 °C, the degradation was more marked than in EVOO stored in dark and at 4 °C, due to the greater influence of external factors on storage conditions. Although chemical–physical characteristics of EVOOs are slightly different depending on provenience and treatment time, the results of this study reveal that storage conditions are fundamental to controlling phenol concentration. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Hydrolytic process that affects oleocanthal and oleacein, leading to the formation of tyrosol and hydroxytyrosol, respectively.</p> Full article ">Figure 2
<p>Oxidative process leads to the formation of oleocanthalic acid from oleocanthal.</p> Full article ">Figure 3
<p>Total phenolic content (TPC) (µg GAE/g EVOO, ppm) in each EVOOs (A, B, and C). Data are expressed as mean ± SD of an experiment performed in triplicate.</p> Full article ">Figure 4
<p>Concentrations of oleocanthal, tyrosol, and oleocanthalic acid (µg of phenolic compound/g of EVOO, ppm) in EVOOs A (<b>a</b>), B (<b>b</b>), and C (<b>c</b>) stored at 4 °C in dark conditions (blue bars) and at room temperature and exposed to daylight (red bars), for 15 months.</p> Full article ">Figure 4 Cont.
<p>Concentrations of oleocanthal, tyrosol, and oleocanthalic acid (µg of phenolic compound/g of EVOO, ppm) in EVOOs A (<b>a</b>), B (<b>b</b>), and C (<b>c</b>) stored at 4 °C in dark conditions (blue bars) and at room temperature and exposed to daylight (red bars), for 15 months.</p> Full article ">
16 pages, 3063 KiB  
Article
Antioxidative Activity Evaluation of High Purity and Micronized Tartary Buckwheat Flavonoids Prepared by Antisolvent Recrystallization
by Yanjie Liu, Xiaoyu Sui, Xiuhua Zhao, Siying Wang and Qilei Yang
Foods 2022, 11(9), 1346; https://doi.org/10.3390/foods11091346 - 5 May 2022
Viewed by 2078
Abstract
Tartary buckwheat, a healthy food, is associated with a reduced risk of certain human chronic diseases. However, the bioactive component flavonoids in Tartary buckwheat have poor solubility and low absorption in vivo. To improve these points, 60.00% Tartary buckwheat total flavonoids (TFs) were [...] Read more.
Tartary buckwheat, a healthy food, is associated with a reduced risk of certain human chronic diseases. However, the bioactive component flavonoids in Tartary buckwheat have poor solubility and low absorption in vivo. To improve these points, 60.00% Tartary buckwheat total flavonoids (TFs) were obtained by ethanol refluxing method, which were purified and micronized by antisolvent recrystallization (ASR) using methanol as a solvent and deionized water as an antisolvent. By using High Performance Liquid Chromatography (HPLC) and electrospray ionized mass spectrometry (ESI-MS), the main flavonoid in pure flavonoids (PF) were rutin (RU), kaempferol-3-O-rutinoside (KA) and quercetin (QU); the content of TF is 99.81% after purification. It is more worthy of our attention that micronized flavonoids contribute more to antioxidant activity because of good solubility. These results provide a theoretical reference for the micronization of other flavonoids. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Chemical structures of Rutin (1), quercetin (2), and kaempferol-3-rutinoside (3).</p> Full article ">Figure 2
<p>Schematic description of antisolvent operation procedure.</p> Full article ">Figure 3
<p>RSM plots—(<b>a</b>) Effect of TF concentration and stirring time; (<b>b</b>) effect of temperature and stirring time; (<b>c</b>) effect of temperature and volume ratio of water and methanol on PF yield; (<b>d</b>) effect of total flavonoids and volume ratio of water and methanol; (<b>e</b>) effect of TF concentration and stirring time on PF purity.</p> Full article ">Figure 4
<p>HPLC chromatogram: reference standards of three flavonoids (blue line), the main flavonoids in PF (red line), and CF (black line).</p> Full article ">Figure 5
<p>The UPLC-MS/MS chromatograms of three compounds (<b>a</b>–<b>c</b>); MS/MS fragmentation of three compounds (<b>d</b>–<b>f</b>) in negative ion mode.</p> Full article ">Figure 6
<p>SEM photos and particle size distribution of sample morphology: (<b>a</b>) CF; (<b>b</b>) PF; (<b>c</b>) particle size distribution.</p> Full article ">Figure 7
<p>(<b>a</b>) Equilibrium solubility of total flavonoids of CF and PF in water, SGF, and SIF (means ± SD, n = 3, * <span class="html-italic">p</span> &lt; 0.05 compared with CF). (<b>b</b>) Fluorescence intensity after treatment at different concentrations of CF and PF over time. (<b>c</b>) The images of HepG2 cells incubated with CF or PF for 50, 100, and 200 μg/mL, which were measured by fluorescence microscope. (<b>d</b>) The residual amount of CF and PF after digestion, and the residual amount of RU in CF with PF after digestion. (means ± SD, n = 3, * <span class="html-italic">p</span> &lt; 0.05 compared with CF, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05 compared with rutin in CF). (<b>e</b>) Dissolution rate of CF and PF after digestion in vitro, and dissolution rate of CF with PF after digestion in vitro (means ± SD, n = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 compared with CF, <sup>#</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>##</sup> <span class="html-italic">p</span> &lt; 0.01 compared with rutin in CF). (<b>f</b>) Cumulative absorption profiles of CF and PF in the intestinal sac model. (means ± SD, n = 3, * <span class="html-italic">p</span> &lt; 0.05 compared with CF).</p> Full article ">Figure 8
<p>DPPH (<b>a</b>) and MDA (<b>b</b>) of CF and PF as they pass through an in vitro human digestion model (means ± SD, n = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 compared with CF).</p> Full article ">
12 pages, 23889 KiB  
Article
Effects of Organic Acids on the Release of Fruity Esters in Water: An Insight at the Molecular Level
by Yu Liu, Hui Xi, Yingjie Fu, Peng Li, Shihao Sun and Yongli Zong
Molecules 2022, 27(9), 2942; https://doi.org/10.3390/molecules27092942 - 4 May 2022
Cited by 10 | Viewed by 2401
Abstract
It is well known that organic acids (OAs) could affect the flavour of fruit juices and beverages. However, the molecular mechanism of aroma release is still unclear. In this study, the effects of citric acid (CA), L-(-)-malic acid (MA) and L-lactic acid (LA) [...] Read more.
It is well known that organic acids (OAs) could affect the flavour of fruit juices and beverages. However, the molecular mechanism of aroma release is still unclear. In this study, the effects of citric acid (CA), L-(-)-malic acid (MA) and L-lactic acid (LA) on the release of six selected esters and their sensory perception were investigated by means of HS-GC-MS analyses and odour detection threshold determination, respectively. Meanwhile, the density functional theory (DFT) calculation was employed to explore the interaction modes between esters and OAs. HS-GC-MS analyses showed that the concentration and the type of OAs regulated the release of esters. The results were basically consistent with the detection threshold change of those esters. The DFT calculation suggested that the main intermolecular interaction was hydrogen bonds, and several esters could form a ternary ring structure with OAs through hydrogen bonds. The interactions can induce the different release behaviours of esters in OAs water solution. The number of carboxyl functional groups in OAs and the spatial conformation of esters appeared to influence the magnitude of the interaction. The above results demonstrated the mechanism of OAs affecting the release of esters and indicated a possible flavour control way by using different OAs and OA concentrations. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Weak interactions between esters and citric acid. (<b>A</b>) C2C3, (<b>B</b>) 2MeC3C2, (<b>C</b>) C4C2, (<b>D</b>) C2C4, (<b>E</b>) 3MeC4C2, (<b>F</b>) C5C2, H-white, C-gray, O-red.</p> Full article ">Figure 2
<p>Weak interactions between esters and malic acid. (<b>A</b>) C2C3, (<b>B</b>) 2MeC3C2, (<b>C</b>) C4C2, (<b>D</b>) C2C4, (<b>E</b>) 3MeC4C2, (<b>F</b>) C5C2, H-white, C-gray, O-red.</p> Full article ">Figure 3
<p>Weak interactions between esters and lactic acid. (<b>A</b>) C2C3, (<b>B</b>) 2MeC3C2, (<b>C</b>) C4C2, (<b>D</b>) C2C4, (<b>E</b>) 3MeC4C2, (<b>F</b>) C5C2, H-white, C-gray, O-red.</p> Full article ">Figure 4
<p><span class="html-italic">R</span> values of esters in 1, 2.5 and 5 g/L of CA solution. Different letters indicate significant difference for each ester (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Relationship between logarithmic value of <span class="html-italic">R</span> values and log <span class="html-italic">P</span> of five retained esters in 1 g/L of CA solution.</p> Full article ">Figure 6
<p><span class="html-italic">R</span> values of esters in 1, 2.5 and 5 g/L of LA solution. Different letters indicate significant difference for each ester (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 7
<p><span class="html-italic">R</span> values of esters in 1, 2.5 and 5 g/L of MA solution. Different letters indicate significant difference for each ester (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 8
<p>The workflow of theoretical calculation.</p> Full article ">
12 pages, 2877 KiB  
Article
Effect of Aucubin-Containing Eye Drops on Tear Hyposecretion and Lacrimal Gland Damage Induced by Urban Particulate Matter in Rats
by Su-Bin Park, Woo Kwon Jung, Hwa-Young Yu, Yong Hwan Kim and Junghyun Kim
Molecules 2022, 27(9), 2926; https://doi.org/10.3390/molecules27092926 - 4 May 2022
Cited by 4 | Viewed by 1917
Abstract
Exposure to particulate matter is a causative factor of dry eye disease. We aimed to investigate the beneficial effect of eye drops containing aucubin on dry eye disease induced by urban particulate matter (UPM). Dry eye was induced in male SD rats (6 [...] Read more.
Exposure to particulate matter is a causative factor of dry eye disease. We aimed to investigate the beneficial effect of eye drops containing aucubin on dry eye disease induced by urban particulate matter (UPM). Dry eye was induced in male SD rats (6 weeks old) by topical exposure to UPM thrice a day for 5 d. Eye drops containing 0.1% aucubin or 0.5% aucubin were topically administered directly into the eye after UPM exposure for an additional 5 d. Tear secretion was evaluated using a phenol red thread tear test and corneal irregularity. The oxidative damage in the lacrimal gland was evaluated using TUNEL and immunohistochemical staining. The topical administration of aucubin significantly attenuated UPM-induced tear hyposecretion (control group: 9.25 ± 0.62 mm, UPM group: 4.55 ± 0.25 mm, 0.1% aucubin: 7.12 ± 0.58 mm, and 0.5% aucubin: 7.88 ± 0.75 mm) and corneal irregularity (control group: 0.00 ± 0.00, UPM group: 3.40 ± 0.29, 0.1% aucubin: 1.80 ± 0.27, and 0.5% aucubin: 1.15 ± 0.27). In addition, aucubin also reduced the UPM-induced apoptotic injury of lacrimal gland cells induced by oxidative stress through the increased expression of HMGB1 and RAGE. These findings indicate that the topical administration of aucubin eye drops showed a beneficial effect against UPM-induced abnormal ocular changes, such as tear hyposecretion and lacrimal gland damage. Therefore, our results reveal the pharmacological activities of aucubin in dry eye disease. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Effects of aucubin eye drops on tear hyposecretion, and corneal irregularity in UPM-induced dry eye rats. (<b>A</b>) The chemical structure of aucubin. (<b>B</b>) The tear volume was measured using a phenol red thread. (<b>C</b>) Representative reflected images of the ring illuminator on the corneal surface. Bar graph indicates the quantitative analysis of the corneal irregularity score. Data shown are mean ± standard deviation (SD) (<span class="html-italic">n</span> = 8). * <span class="html-italic">p</span> &lt; 0.05 vs. Control group, # <span class="html-italic">p</span> &lt; 0.05 vs. UPM group. Control group: Control, UPM-treated group: UPM, UPM + 0.1% aucubin-eye-drop-treated group: 0.1% aucubin, and UPM + 0.5% aucubin-eye-drop-treated group: 0.5% aucubin.</p> Full article ">Figure 2
<p>Effect of aucubin eye drops on ROS generation in lacrimal glands of UPM-induced dry eye rats. (<b>A</b>) Total levels of ROS in the lacrimal gland. (<b>B</b>) Immunohistochemical staining for 8-OHdG. Bar graph indicates the quantification of the 8-OHdG signal intensity. Data shown are mean ± SD (<span class="html-italic">n</span> = 8). * <span class="html-italic">p</span> &lt; 0.05 vs. Control group, # <span class="html-italic">p</span> &lt; 0.05 vs. UPM group. Control group: Control, UPM-treated group: UPM, UPM + 0.1% aucubin-eye-drop-treated group: 0.1% aucubin, and UPM + 0.5% aucubin-eye-drop-treated group: 0.5% aucubin.</p> Full article ">Figure 3
<p>Effect of aucubin eye drops on apoptosis in lacrimal glands of UPM-induced dry eye rats. Representative images for TUNEL staining are presented. The numbers of TUNEL-positive cells were counted. Data shown are mean ± SD (<span class="html-italic">n</span> = 8). * <span class="html-italic">p</span> &lt; 0.05 vs. Control group, # <span class="html-italic">p</span> &lt; 0.05 vs. UPM group. Control group: Control, UPM-treated group: UPM, UPM + 0.1% aucubin-eye-drop-treated group: 0.1% aucubin, and UPM + 0.5% aucubin-eye-drop-treated group: 0.5% aucubin.</p> Full article ">Figure 4
<p>Effect of aucubin eye drops on the expression of HMGB1 in lacrimal glands of UPM-induced dry eye rats. (<b>A</b>) Representative images for immunohistochemical staining for HMGB1. (<b>B</b>) Bar graph indicates the quantification of the HMGB1 signal intensity. (<b>C</b>) Relative mRNA levels of HMGB1 were assessed using a quantitative RT-PCR. Data shown are mean ± SD (<span class="html-italic">n</span> = 8). * <span class="html-italic">p</span> &lt; 0.05 vs. Control group, # <span class="html-italic">p</span> &lt; 0.05 vs. UPM group. Control group: Control, UPM-treated group: UPM, UPM + 0.1% aucubin-eye-drop-treated group: 0.1% aucubin, and UPM + 0.5% aucubin-eye-drop-treated group: 0.5% aucubin.</p> Full article ">Figure 5
<p>Effect of aucubin eye drops on the expression of RAGE in lacrimal glands of UPM-induced dry eye rats. (<b>A</b>) Representative images for immunohistochemical staining for RAGE. (<b>B</b>) Bar graph indicates the quantification of the RAGE signal intensity. (<b>C</b>) Relative mRNA levels of RAGE were assessed using a quantitative RT-PCR. Data shown are mean ± SD (<span class="html-italic">n</span> = 8). * <span class="html-italic">p</span> &lt; 0.05 vs. Control group, # <span class="html-italic">p</span> &lt; 0.05 vs. UPM group. Control group: Control, UPM-treated group: UPM, UPM + 0.1% aucubin-eye-drop-treated group: 0.1% aucubin, and UPM + 0.5% aucubin-eye-drop-treated group: 0.5% aucubin.</p> Full article ">Figure 6
<p>Experimental design for the animal study.</p> Full article ">
15 pages, 3109 KiB  
Article
Pro-Angiogenic Effects of Natural Antioxidants Extracted from Mango Leaf, Olive Leaf and Red Grape Pomace over Endothelial Colony-Forming Cells
by Ismael Sánchez-Gomar, Josefa Benítez-Camacho, Cristina Cejudo-Bastante, Lourdes Casas, Rafael Moreno-Luna, Casimiro Mantell and Mª Carmen Durán-Ruiz
Antioxidants 2022, 11(5), 851; https://doi.org/10.3390/antiox11050851 - 27 Apr 2022
Cited by 11 | Viewed by 3170
Abstract
Cardiovascular diseases remain the leading cause of death worldwide, mainly triggered by the formation of atherosclerotic plaques that reduce blood flow. Angiogenic cell therapy based on endothelial colony forming cells (ECFCs) constitutes a promising alternative to promote vascular revascularization; however, under the oxidative [...] Read more.
Cardiovascular diseases remain the leading cause of death worldwide, mainly triggered by the formation of atherosclerotic plaques that reduce blood flow. Angiogenic cell therapy based on endothelial colony forming cells (ECFCs) constitutes a promising alternative to promote vascular revascularization; however, under the oxidative environment that prevails in ischemic areas, these cells become impaired. Thus, it is necessary to investigate strategies to enhance their regenerative properties. Antioxidant substances, such as polyphenols, have been shown to be useful for this purpose. In the current study we evaluated the potential of mango leaves, olive leaves and red grape pomace extracts, rich in polyphenols, to promote ECFC reparative effects. For this, aqueous and ethanolic extracts of the aforementioned raw materials were obtained by pressurized liquid extraction (PLE). After evaluating the polyphenol content and the antioxidant activity, in vitro assays were carried out, and we found that ethanolic extracts at low concentrations improved angiogenic capacities of ECFCs and reduced proliferation, apoptosis, and the inflammatory response of these cells. Overall, mango leaves ethanolic extract provided the most promising results, but all three extracts ameliorated the functionality of ECFCs. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Chemical and functional characterization of the extracts obtained by PLE. (<b>A</b>) Graphical representation of global yield of the aqueous and ethanolic extracts of mango leaf, olive leaf, and red grape pomace expressed as g/100 g dry extract (%). (<b>B</b>) Total phenolic content expressed as g/100 g dry extract. (<b>C</b>) Antioxidant activity represented by the efficient concentration (EC<sub>50</sub>) (µg/mL). (<b>D</b>) Antioxidant activity index (AAI) (µg DPPH/µg extract)<b>.</b> Values are represented as the mean ± SD. *** <span class="html-italic">p</span>-value &lt; 0.001.</p> Full article ">Figure 2
<p>Effect of selected extracts on the angiogenic process of ECFCs. (<b>A</b>) Representative images of the reticular structures formed by ECFCs after 24, 48 and 72 h incubation in basal medium, with the inhibitor sulforaphane (15 mM), the activator FGF (35 ng/mL), or with mango (4.99 µg/mL), olive (46.67 µg/mL), and grape (16.67 µg/mL) ethanolic extracts. (<b>B</b>) Graphical representation of the changes seen for the number of meshes, number of segments, and total length of the segments analyzed in ECFCs treated with mango leaves, olive leaves, and red grape pomace ethanolic extracts after 24, 48, and 72 h. (<b>C</b>) Temporal evolution of the number of meshes in ECFCs treated with the selected extracts. * <span class="html-italic">p</span> &lt; 0.05; *** <span class="html-italic">p</span> &lt; 0.001.</p> Full article ">Figure 3
<p>Effect of the selected extracts on ECFC proliferation and differentiation. (<b>A</b>) Representative images of Ki67 and vWF staining of ECFCs treated 48 h with the ethanolic extracts of mango (4.99 μg/mL) and olive (46.67 μg/mL) leaves, and red grape pomace (11.67 μg/mL). (<b>B</b>) Box and whisker diagrams of the effect of the selected extracts on the proliferation and differentiation of ECFCs after 48 h of treatment. * <span class="html-italic">p</span>-value: * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p> Full article ">Figure 4
<p>Effect of the selected extracts on the apoptosis of ECFCs. (<b>A</b>) Representative dot-plots of mango leaves, olive leaves, and red grape pomace extracts effect on ECFCs apoptosis after 24 and 48 h of treatment. (<b>B</b>) Graphical representation of the effect of mango leaves (4.99 μg/mL), olive leaves (46.67 μg/mL), and red grape pomace (11.67 μg/mL) on early and late apoptosis in ECFCs after 24 and 48 h of treatment. * <span class="html-italic">p</span>-value: * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001.</p> Full article ">Figure 5
<p>Anti-inflammatory effect of selected extracts. (<b>A</b>) Representative histograms of the effect of treatment with TNFα alone and TNFα with the mango (4.99 μg/mL), olive (46.67 μg/mL), and grape (16.67 μg/mL) ethanolic extracts on the expression of the adhesion molecules E-selectin and VCAM-1, at 4 h. (<b>B</b>) Box and whisker diagrams of the effect of selected extracts on the expression of adhesion molecules. MFI: Mean Fluorescence Signal. ### Significant differences with respect to the control condition (-TNFα). * Significant differences between the conditions. * <span class="html-italic">p</span> &lt; 0.05; *** <span class="html-italic">p</span> &lt; 0.001.</p> Full article ">
13 pages, 913 KiB  
Article
Evaluation of Different Black Mulberry Fruits (Morus nigra L.) Based on Phenolic Compounds and Antioxidant Activity
by Ri-Si Wang, Pan-Hao Dong, Xi-Xiang Shuai and Ming-Shun Chen
Foods 2022, 11(9), 1252; https://doi.org/10.3390/foods11091252 - 26 Apr 2022
Cited by 29 | Viewed by 3102
Abstract
This study evaluated thirteen different black mulberry fruits (Morus nigra L.) grown in the Guangdong region in order to select the best cultivar for health benefits and commercial applications. The phenolic compounds were identified and quantified using UPLC-ESI-MS/MS. The antioxidant activity was [...] Read more.
This study evaluated thirteen different black mulberry fruits (Morus nigra L.) grown in the Guangdong region in order to select the best cultivar for health benefits and commercial applications. The phenolic compounds were identified and quantified using UPLC-ESI-MS/MS. The antioxidant activity was evaluated by three in vitro methods. Significant differences among samples were found regarding total soluble solids (6.20–15.83 °Brix), titratable acidity (5.82–48.49 mg CA/g), total phenolic contents (10.82–27.29 mg GAE/g), total flavonoid contents (1.21–2.86 mg RE/g) and total anthocyanin contents (2.91–11.86 mg CE/g). Fifty-five different phenolic compounds were identified, of which fifteen were reported in mulberry for the first time, but only forty-six of them were quantitated. The DPPH radical scavenging activity, ABTS radical scavenging activity and ferric ion-reducing antioxidant power varied significantly among the samples. Overall, cultivars with better combinations of phenolic compounds and antioxidant activity were Qiong46 (M-2), Yuebanguo (M-4) and Heizhenzhu (M-10), which were recommended for commercial cultivation. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Antioxidant activities of different mulberry samples measured by (<b>A</b>) DPPH, (<b>B</b>) ABTS and (<b>C</b>) FRAP assays. Values in the different letters are significant differences (<span class="html-italic">p</span> &lt; 0.05). M-1, Yichuanhong; M-2, Qiong46; M-3, Guosang8632; M-4, Yuebanguo; M-5, Yueshen28; M-6, Yueshen74; M-7, D10; M-8, Jinqiang63; M-9, Yunguo1; M-10, Heizhenzhu; M-11, Xuanguo1; M-12, Guiyou12; M-13, Shansang.</p> Full article ">Figure 2
<p>(<b>A</b>) PC scores plot of the mulberry samples; (<b>B</b>) loadings plot of the mulberry samples. Salad, salicylic acid; TraCin, trans-cinnamic acid; Proad, protocatechuic acid; Chlad, chlorogenic acid; Cryad, cryptochlorogenic acid; Ferad, ferulic acid; Sinad, sinapic acid; Nar, narcissin; Aro, aromadendrin; Tax, taxifolin; Quegal, quercetin-3-galactoside; Isoglu, isorhamnetin-3-O-glucoside; Kaerut, kaempferol-3-O-rutinoside; Rut, rutin; But, butein; Eri, eriodictyol; Pru, prunin; Hes, hesperidin; Tri, trilobatin; Lut, luteolin; Mor, morin; Que, quercetin; Cat, catechin; Epi, epicatechin; Pel, pelargonidin; Delglu, delphinidin-3-glucoside; Cya, cyanidin-3,5-diglucoside; Cyarut, cyanidin-3-O-rutinoside; ProB1, procyanidin B1; ProB3, procyanidin B3; Fra, fraxin; Res, resveratrol.</p> Full article ">Figure 3
<p>Hierarchical cluster analysis of the mulberry samples.</p> Full article ">
14 pages, 3026 KiB  
Article
Extract from Black Soybean Cultivar A63 Extract Ameliorates Atopic Dermatitis-like Skin Inflammation in an Oxazolone-Induced Murine Model
by Banzragch Dorjsembe, Chu Won Nho, Yongsoo Choi and Jin-Chul Kim
Molecules 2022, 27(9), 2751; https://doi.org/10.3390/molecules27092751 - 25 Apr 2022
Cited by 7 | Viewed by 2789
Abstract
Black soybean has been used in traditional medicine to treat inflammatory diseases, cancer, and diabetes and as a nutritional source since ancient times. We found that Korean black soybean cultivar A63 has more cyanidin-3-O-glucoside, (C3G), procyanidin B2 (PB2), and epicatechin (EPC) [...] Read more.
Black soybean has been used in traditional medicine to treat inflammatory diseases, cancer, and diabetes and as a nutritional source since ancient times. We found that Korean black soybean cultivar A63 has more cyanidin-3-O-glucoside, (C3G), procyanidin B2 (PB2), and epicatechin (EPC) contents than other cultivars and has beneficial effects on cell viability and anti-oxidation. Given the higher concentration of anthocyanidins and their strong anti-oxidant activity, we predicted that A63 extract could relieve inflammatory disease symptoms, including those of atopic dermatitis (AD). Here, we evaluated the anti-AD activity of A63 extract in an oxazolone (OXA)-induced mouse model. A63 extract treatment significantly reduced epidermal thickness and inflammatory cell infiltration, downregulated the expression of AD gene markers, including Interleukin (IL)-4 and IL-5, and restored damaged skin barrier tissues. Furthermore, A63 extract influenced the activation of the signal transducer and activator of transcription (STAT) 3 and STAT6, extracellular regulatory kinase (ERK), and c-Jun N-terminal kinase (JNK) signaling pathways, which play a crucial role in the development of AD. Altogether, our results suggest that A63 can ameliorate AD-like skin inflammation by inhibiting inflammatory cytokine production and STAT3/6 and Mitogen-activated protein kinase (MAPK) signaling and restoring skin barrier function. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>HPLC analysis of ST and A63 extract and its major compounds. Typically extracted ion chromatogram of three major anti-oxidants, Cyanidin-3-<span class="html-italic">O</span>-Glucoside (C3G), procyanidin B2 (PB2), and epicatechin (EP), in Korean black soybean cultivar ST (<b>A</b>) and A63 (<b>B</b>).</p> Full article ">Figure 2
<p>A63 extract attenuates OXA-induced AD-like skin inflammation in mice. Histological features were analyzed by H&amp;E staining for epidermal thickness (<b>A</b>), Toluidine Blue staining for mast cells (<b>B</b>), and macrophages with surface marker F4/80 proteins by IHC (<b>C</b>). Additionally, measurement of epidermal thickness (<b>D</b>) and counting of mast cell (<b>E</b>) and F4/80 positive macrophages (<b>F</b>) were performed. All representative images were obtained at 20× magnification, and epidermal thickness and the number of mast cells and F4/80 macrophages were examined with an Olympus DP27 microscope (Olympus Cell Sens Standard software). Results are expressed as mean ± standard deviation (n = 5, # <span class="html-italic">p</span> &lt; 0.05, ### <span class="html-italic">p</span> &lt; 0.001 versus control group, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 versus 1% OXA group, scale bar = 50 µm). OXA, oxazolone; AD, atopic dermatitis; H&amp;E, hematoxylin and eosin; IHC, immunohistochemistry.</p> Full article ">Figure 3
<p>A63 extract application enhanced the restoration of normal skin integrity. Immunohistochemical analysis of ears in an OXA-induced AD murine model. A63 extract treatment increased expression of skin barriers components, filaggrin (<b>A</b>, FLG), keratin 14 (<b>B</b>, K14) and loricrin (<b>C</b>, LOR) in OXA-triggered mice similar to the Dex group (scale bar = 50 µm). OXA, oxazolone; AD, atopic dermatitis; Dex, dexamethasone.</p> Full article ">Figure 4
<p>A63 extract suppresses the expression of atopic dermatitis hallmark genes in mouse tissue. qPCR analysis of atopic marker expression in mouse skin. The elevated production of atopic cytokines, IL-4 (<b>A</b>), IL-5 (<b>B</b>), CCL17 (<b>C</b>), and CCL26 (<b>D</b>), in OXA-treated groups was modestly suppressed by A63 extract in ear tissue. Results are expressed as mean ± standard deviation (### <span class="html-italic">p</span> &lt; 0.001 versus control group, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 versus 1% OXA group). OXA, oxazolone.</p> Full article ">Figure 5
<p>The activation of MAPK and STAT signaling was hindered by treatment with A63 extract. MAPK and STAT signaling pathways play crucial roles in the maturation of naive T cells, recruitment of immune cells, and secretion of atopic cytokines. A63 extract (1%) abolished the activation of MAPK signaling in atopic mice (<b>A</b>). STAT3/6 protein phosphorylation was suppressed dose-dependently upon A63 extract treatment (<b>B</b>).</p> Full article ">Figure 6
<p>A63 extract prevents eosinophil infiltration toward the inflammatory site via CCL26 expression downregulation and the inactivation of JAK1/STAT6 signaling. qPCR analysis of CCL26 expression in IL-4/IL-13-induced HS68 cells (<b>A</b>). Western blot results of IL-4/IL-13-induced HS68 cells (<b>B</b>). Results are expressed as mean ± standard deviation (### <span class="html-italic">p</span> &lt; 0.001 versus control group, *** <span class="html-italic">p</span> &lt; 0.001 versus IL-4/13 treated group).</p> Full article ">
11 pages, 2793 KiB  
Article
Comparative Analysis of Phytochemical Profiles and Antioxidant Activities between Sweet and Sour Wampee (Clausena lansium) Fruits
by Xiaoxiao Chang, Yutong Ye, Jianping Pan, Zhixiong Lin, Jishui Qiu, Cheng Peng, Xinbo Guo and Yusheng Lu
Foods 2022, 11(9), 1230; https://doi.org/10.3390/foods11091230 - 25 Apr 2022
Cited by 11 | Viewed by 2162
Abstract
As a local medicine and food, wampee fruit, with abundant bioactive compounds, is loved by local residents in Southern China. Titratable acid (TA), total sugar (TS), and total phenolic and flavonoid contents were detected, and phytochemical profiles and cellular antioxidant activities were analyzed [...] Read more.
As a local medicine and food, wampee fruit, with abundant bioactive compounds, is loved by local residents in Southern China. Titratable acid (TA), total sugar (TS), and total phenolic and flavonoid contents were detected, and phytochemical profiles and cellular antioxidant activities were analyzed by the HPLC and CAA (cellular antioxidant activity) assay in five sweet wampee varieties and five sour wampee varieties. Results showed that the average TS/TA ratio of sweet wampee varieties was 29 times higher than sour wampee varieties, while TA content was 19 times lower than sour wampee varieties. There were much lower levels of total phenolics, flavonoids, and antioxidant activities in sweet wampee varieties than those in sour wampee varieties. Eight phytochemicals were detected in sour wampee varieties, including syringin, rutin, benzoic acid, 2-methoxycinnamic acid, kaempferol, hesperetin, nobiletin, and tangeretin, while just four of them were detected in sweet wampee varieties. Syringin was the only one that was detected in all the sour wampee varieties and was not detected in all sweet wampee varieties. Correlation analysis showed significant positive correlations between TA with phenolics, flavonoids, and total and cellular (PBS wash) antioxidant activities, while there were significant negative correlations between TS/TA with phenolic and cellular (no PBS wash) antioxidant activities. This suggested that the content of titratable acid in wampee fruit might have some relationship with the contents of phenolics and flavonoids. Sour wampee varieties should be paid much attention by breeders for their high phytochemical contents and antioxidant activities for cultivating germplasms with high health care efficacy. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The varieties of wampee fruits. THP: TianHuangPi; ZFHP: ZaoFengHuangPi; CCTP: CongChengTianPi; LTDH: LuTianDuHe; TXTP: TaXiaTianPi; M3H: Min3Hao; JZP: JiZiPi; M4H: Min4Hao; TXSP: TaXiaSuanPi; JXHP: JiXinHuangPi.</p> Full article ">Figure 2
<p>Total phenolic contents of five sweet and five sour wampee varieties. Bars with no letters in common are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Total flavonoid contents of five sweet and five sour wampee varieties. Bars with no letters in common are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Total antioxidant activities of the five sweet and five sour wampee varieties obtained by the ORAC assay. Bars with no letters in common are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>The cellular activities of the five sweet and five sour wampee varieties obtained by the ORAC assay. Bars with no letters in common are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 6
<p>Heat map of the correlation analysis among titratable acid, total sugar, total phenolics, total flavonoids, phytochemical components, and antioxidant activities. TA: titratable acid; TS: total sugar; TSTA: total sugar/titratable acid; TPC: total phenolic content; TFC: total flavonoid content; ORAC: the oxygen radical absorbance capacity; CAANW: CAA value without PBS wash; CAAW: CAA value with PBS wash; SY: syringin; RU: rutin; BE: benzoic acid; ME: 2-methoxycinnamic acid; KA: kaempferol; HE: hesperetin; NO: nobiletin; TG: tangeretin. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p> Full article ">
11 pages, 1716 KiB  
Article
Purification and Identification of Novel Myeloperoxidase Inhibitory Antioxidant Peptides from Tuna (Thunnas albacares) Protein Hydrolysates
by Bingna Cai, Peng Wan, Hua Chen, Jingtong Huang, Ziqing Ye, Deke Chen and Jianyu Pan
Molecules 2022, 27(9), 2681; https://doi.org/10.3390/molecules27092681 - 21 Apr 2022
Cited by 17 | Viewed by 2370
Abstract
Antioxidative peptides that inhibit myeloperoxidase (MPO) enzyme activity can effectively defend against oxidative stress damage. The antioxidant peptides from tuna protein were produced using alcalase hydrolysis and purified by ultrafiltration and Sephadex G-15, and the fractions with the highest free radicals scavenging ability [...] Read more.
Antioxidative peptides that inhibit myeloperoxidase (MPO) enzyme activity can effectively defend against oxidative stress damage. The antioxidant peptides from tuna protein were produced using alcalase hydrolysis and purified by ultrafiltration and Sephadex G-15, and the fractions with the highest free radicals scavenging ability and oxygen radical absorbance capacity (ORAC) values were sequenced using HPLC–MS/MS. Fifty-five peptide sequences were identified, 53 of which were successfully docked into MPO. The representative peptide ACGSDGK had better antioxidant activity and inhibition of MPO chlorination and peroxidation than the reference peptide hLF1-11. The docking model further showed intense molecular interactions between ACGSDGK and MPO, including hydrogen bonds, charge, and salt bridge interactions, which occluded the active site and blocked the catalytic activity of MPO. These results suggested that the antioxidant peptide ACGSDGK has the potential to inhibit oxidative stress and alleviate inflammation in vivo because of its inhibitory effect on the MPO enzyme. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Hydrolysis degree (<b>A</b>) and antioxidant activity (<b>B</b>) of hydrolysate at different enzymolysis times. Different lowercase letters on the bar indicated significant difference (<span class="html-italic">p</span> &lt; 0.05) between groups.</p> Full article ">Figure 2
<p>The antioxidant activity of ultrafiltration fractions (<b>A</b>). Fraction &lt;3 kDa with the strongest antioxidant activity was separated by Sephadex G-15 gel (<b>B</b>). Seven fractions were collected, and their antioxidant activity was determined by ABTS and ORAC assays (<b>C</b>). Different lowercase letters on the bar indicated significant difference (<span class="html-italic">p</span> &lt; 0.05) between groups.</p> Full article ">Figure 3
<p>Three-dimensional and 2D structure graphs of the molecular interactions between peptide ACGSDGK (<b>A</b>) and MPO, as well as between peptide KFCSGHA (<b>B</b>) and MPO.</p> Full article ">
20 pages, 2626 KiB  
Article
Effect of Exogenous Melatonin Application on the Grain Yield and Antioxidant Capacity in Aromatic Rice under Combined Lead–Cadmium Stress
by Ye Jiang, Suihua Huang, Lin Ma, Leilei Kong, Shenggang Pan, Xiangru Tang, Hua Tian, Meiyang Duan and Zhaowen Mo
Antioxidants 2022, 11(4), 776; https://doi.org/10.3390/antiox11040776 - 13 Apr 2022
Cited by 23 | Viewed by 4346
Abstract
This study aimed to determine the mechanism of exogenous melatonin application in alleviating the combined Pb and Cd (Pb-Cd) toxicity on aromatic rice (Oryza sativa L.). In this study, a pot experiment was conducted; two aromatic rice varieties, Yuxiangyouzhan and Xiangyaxiangzhan, were [...] Read more.
This study aimed to determine the mechanism of exogenous melatonin application in alleviating the combined Pb and Cd (Pb-Cd) toxicity on aromatic rice (Oryza sativa L.). In this study, a pot experiment was conducted; two aromatic rice varieties, Yuxiangyouzhan and Xiangyaxiangzhan, were selected, and sprays using 50, 100, 200, and 400 μmol L−1 melatonin (denoted as S50, S100, S200, and S400) and irrigation using 100, 300, and 500 μmol L−1 melatonin (denoted as R100, R300, and R500) were also selected. The results showed that, under the S50, S100, and S200 treatments, the Pb content of aromatic rice grain decreased, and the grain yield increased significantly. Moreover, the application of exogenous melatonin significantly reduced the accumulation of H2O2 in rice leaves at maturity under Cd–Pb stress and reduced the MDA content in Xiangyaxiangzhan leaves. In addition, the microbial community structure changed significantly under S50 and R300 treatments. Some pathways, such as the synthesis of various amino acids and alanine, aspartate, and glutamate metabolism, were regulated by S50 treatment. Overall, melatonin application improved aromatic rice grain yield while reducing heavy metal accumulation by regulating the antioxidant capacity and metabolites in aromatic rice plants and altering the physicochemical properties and microbial community structures of the soil. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The average monthly temperature, humidity, and sunshine hours during the experiment period in 2019.</p> Full article ">Figure 2
<p>Effects of exogenous melatonin application on GSH content in Yuxiangyouzhan (<b>a</b>) and Xiangyaxiangzhan (<b>b</b>); ASA content in Yuxiangyouzhan (<b>c</b>) and Xiangyaxiangzhan (<b>d</b>); MTs content in Yuxiangyouzhan (<b>e</b>) and Xiangyaxiangzhan (<b>f</b>) at the heading stage (HS), 15 day after HS, and the maturity stage (MS). Means sharing similar letters indicate no significant difference at <span class="html-italic">p</span> &lt; 0.05 according to the LSD test.</p> Full article ">Figure 3
<p>Effects of exogenous melatonin application on SOD activity in Yuxiangyouzhanand (<b>a</b>) and Xiangyaxiangzhan (<b>b</b>); POD activity in Yuxiangyouzhan (<b>c</b>) and Xiangyaxiangzhan (<b>d</b>); CAT activity in Yuxiangyouzhan (<b>e</b>) and Xiangyaxiangzhan (<b>f</b>); MDA content in Yuxiangyouzhan (<b>g</b>) and Xiangyaxiangzhan (<b>h</b>); and H<sub>2</sub>O<sub>2</sub> content in Yuxiangyouzhan (<b>i</b>) and Xiangyaxiangzhan (<b>j</b>) at the heading stage (HS), 15 day after HS, and the MS. Means sharing similar letters indicate no significant difference at <span class="html-italic">p</span> &lt; 0.05 according to the LSD test.</p> Full article ">Figure 3 Cont.
<p>Effects of exogenous melatonin application on SOD activity in Yuxiangyouzhanand (<b>a</b>) and Xiangyaxiangzhan (<b>b</b>); POD activity in Yuxiangyouzhan (<b>c</b>) and Xiangyaxiangzhan (<b>d</b>); CAT activity in Yuxiangyouzhan (<b>e</b>) and Xiangyaxiangzhan (<b>f</b>); MDA content in Yuxiangyouzhan (<b>g</b>) and Xiangyaxiangzhan (<b>h</b>); and H<sub>2</sub>O<sub>2</sub> content in Yuxiangyouzhan (<b>i</b>) and Xiangyaxiangzhan (<b>j</b>) at the heading stage (HS), 15 day after HS, and the MS. Means sharing similar letters indicate no significant difference at <span class="html-italic">p</span> &lt; 0.05 according to the LSD test.</p> Full article ">Figure 4
<p>Rarefaction curve (<b>a</b>); histogram of species distribution (<b>b</b>); PCA plot of fungal communities in 18 soil samples under the XCK, XS50, YCK, YR300 treatments (<b>c</b>); RDA considering the fungal relative abundance at the operational taxonomic unit (OTU) level; SOM, soil total N, P, and K contents, Cd and Pb concentrations, and pH (<b>d</b>). XCK: Xiangyaxiangzhan growing under Cd–Pb stress without melatonin treatment; XR300: Xiangyaxiangzhan growing under Cd–Pb stress and irrigated with 300 μmol L<sup>−1</sup> exogenous melatonin; XS50: Xiangyaxiangzhan growing under Cd–Pb stress and sprayed with 50 μmol L<sup>−1</sup> exogenous melatonin; YCK: Yuxiangyouzhan growing under Cd–Pb stress without melatonin treatment; YR300: Yuxiangyouzhan growing under Cd–Pb stress and irrigated with 300 μmol L<sup>−1</sup> exogenous melatonin; YS50: Yuxiangyouzhan growing under Cd–Pb stress and sprayed with 50 μmol L<sup>−1</sup> exogenous melatonin.</p> Full article ">Figure 5
<p>Overview of the enriched KEGG pathways. 1: Alanine, aspartate, and glutamate metabolism; 2: Arginine biosynthesis; 3: Aminoacyl-tRNA biosynthesis; 4: Nicotinate and nicotinamide metabolism; 5: Phenylpropanoid biosynthesis; 6: Citrate cycle (TCA cycle); 7: Butanoate metabolism; 8: Arginine and proline metabolism; 9: C5-Branched dibasic acid metabolism; 10: Valine, leucine, and isoleucine biosynthesis; 11: Pantothenate and CoA biosynthesis; 12: Glyoxylate and dicarboxylate metabolism; 13: Carbon fixation in photosynthetic organisms; 14: Pyrimidine metabolism; 15: Isoquinoline alkaloid biosynthesis; 16: Starch and sucrose metabolism; 17: beta-Alanine metabolism; 18: Glycine, serine, and threonine metabolism; 19: Vitamin B6 metabolism; 20: Betalain biosynthesis.</p> Full article ">Figure 6
<p>Multivariate pattern recognition analysis of metabolomics. (<b>a</b>) PCA plot of metabolites under XCK, XS50, YCK, and YS50 treatments; (<b>b</b>) PLS-DA plot of metabolites under XCK, XS50, YCK, and YS50 treatments; (<b>c</b>) VIP score of metabolites under XCK, XS50, YCK, and YS50 treatments; (<b>d</b>) PCA plot of metabolites under XCK and XS50 treatments; (<b>e</b>) PLS-DA plot of metabolites under XCK and XS50 treatments; (<b>f</b>) VIP score of metabolites under XCK and XS50 treatments; (<b>g</b>) PCA plot of metabolites under YCK and YS50 treatments; (<b>h</b>) PLS-DA plot of metabolites under YCK and YS50 treatments; and (<b>i</b>) VIP score of metabolites under YCK and YS50 treatments.</p> Full article ">Figure 7
<p>(<b>a</b>) Venn diagram showing the numbers of differential metabolites in XCK vs. XS50, XCK vs. YCK, XS50 vs. YS50, and YCK vs. YS50 comparisons. (<b>b</b>) Significant differential metabolites in XCK vs. XS50, XCK vs. YCK, XS50 vs. YS50, and YCK vs. YS50.</p> Full article ">
13 pages, 1364 KiB  
Article
Components of Banisteriopsis caapi, a Plant Used in the Preparation of the Psychoactive Ayahuasca, Induce Anti-Inflammatory Effects in Microglial Cells
by Beatriz Werneck Lopes Santos, Daniel Carneiro Moreira, Tatiana Karla dos Santos Borges and Eloisa Dutra Caldas
Molecules 2022, 27(8), 2500; https://doi.org/10.3390/molecules27082500 - 13 Apr 2022
Cited by 9 | Viewed by 4468
Abstract
Banisteriopsis caapi is used to prepare the psychoactive beverage ayahuasca, and both have therapeutic potential for the treatment of many central nervous system (CNS) conditions. This study aimed to isolate new bioactive compounds from B. caapi extract and evaluate their biological activity, and [...] Read more.
Banisteriopsis caapi is used to prepare the psychoactive beverage ayahuasca, and both have therapeutic potential for the treatment of many central nervous system (CNS) conditions. This study aimed to isolate new bioactive compounds from B. caapi extract and evaluate their biological activity, and that of the known β-carboline components of the plant (harmine, harmaline, and tetrahydroharmine), in BV-2 microglial cells, the in vivo activation of which is implicated in the physiopathology of CNS disorders. B. caapi extract was fractionated using semipreparative liquid chromatography (HPLC-DAD) and the exact masses ([M + H]+m/z) of the compounds in the 5 isolated fractions were determined by high-resolution LC-MS/MS: F1 (174.0918 and 233.1289), F2 (353.1722), F3 (304.3001), F4 (188.1081), and F5 (205.0785). Harmine (75.5–302 µM) significantly decreased cell viability after 2 h of treatment and increased the number of necrotic cells and production of reactive oxygen species at equal or lower concentrations after 24 h. F4 did not impact viability but was also cytotoxic after 24 h. Most treatments reduced proinflammatory cytokine production (IL-2, IL-6, IL-17, and/or TNF), especially harmaline and F5 at 2.5 µM and higher concentrations, tetrahydroharmine (9.3 µM and higher), and F5 (10.7 µM and higher). The results suggest that the compounds found in B. caapi extract have anti-inflammatory potential that could be explored for the development of treatments for neurodegenerative diseases. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Structures of the main β-carbolines found in <span class="html-italic">Banisteriopsis caapi</span>.</p> Full article ">Figure 2
<p>HPLC-DAD chromatogram of a <span class="html-italic">Banisteriopsis caapi</span> methanolic extract, at 230 nm. Putative identification of procyanidin and epicatechin (Samoylenko et al., 2010) and harmalinic acid (Hashimoto, 1975). <span class="html-italic">m</span>/<span class="html-italic">z</span> refers to the protonated ion ([M + H]<sup>+</sup>) in the UHPLC-TripleTOF 5600+. THH: tetrahydroharmine.</p> Full article ">Figure 3
<p>Viability of BV-2 cells treated at different concentrations for 2 h with <span class="html-italic">Banisteriopsis caapi</span> extract (BC, (<b>A</b>)), the isolated fractions F1 to F5 (<b>B</b>–<b>F</b>), and β-carbolines (<b>G</b>–<b>I</b>). Values are presented as mean ± SEM (<span class="html-italic">n</span> = 3, except for control, <span class="html-italic">n</span> = 6). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared to cells with no treatment (control).</p> Full article ">Figure 4
<p>Cell count (left) and ROS production (right) after 24 h in BV-2 cells treated with <span class="html-italic">Banisteriopsis caapi</span> extract fractions F2 (<b>A</b>,<b>B</b>) and F4 (<b>C</b>,<b>D</b>) and harmine (<b>E</b>,<b>F</b>). Values are presented as mean ± SEM (<span class="html-italic">n</span> = 4–5); * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared to cells with no treatment (control). Viable + early apoptotic and necrotic cells in all treatments is equal to 100%.</p> Full article ">
17 pages, 2769 KiB  
Article
Oriented Deep Eutectic Solvents as Efficient Approach for Selective Extraction of Bioactive Saponins from Husks of Xanthoceras sorbifolia Bunge
by Jinteng Cao, Guangwei Wu, Lei Wang, Fuliang Cao, Yan Jiang and Linguo Zhao
Antioxidants 2022, 11(4), 736; https://doi.org/10.3390/antiox11040736 - 8 Apr 2022
Cited by 11 | Viewed by 3320
Abstract
The husks of Xanthoceras sorbifolia Bunge (X. sorbifolia), as by-products of industrial production, have brought a severe burden to the environment and caused an enormous waste of resources. Bioactive triterpenoid saponins are rich in the husks. To reuse the husks and [...] Read more.
The husks of Xanthoceras sorbifolia Bunge (X. sorbifolia), as by-products of industrial production, have brought a severe burden to the environment and caused an enormous waste of resources. Bioactive triterpenoid saponins are rich in the husks. To reuse the husks and gain high-quality saponin products, saponin-oriented deep eutectic solvents (DESs), as an efficient and selective extraction strategy from X. sorbifolia husks, were designed for the first time. The enhancement of the extraction rate was investigated by screening solvents from acidic DESs and response surface methodology (RSM) optimization. As a result, the tetrapropylammonium bromide-lactic acid (TPMBr-La) was the most efficient DESs, with an extraction efficiency of up to 135% higher than 70% ethanol. A maximum extraction rate of 72.11 ± 0.61 mg Re/g dw was obtained under the optimized parameters. Scanning electron microscope graphs revealed that damage to the microstructure caused by DESs enhanced the extraction efficiency. Moreover, the recovery of total saponins with D101 macroporous resin was consistent with the pseudo-second-order kinetic model. Seven saponins were also identified by HPLC-MS analysis. Finally, TPMBr-La extracts exhibited 92.30 ± 1.10% DPPH radical scavenging rate at 100 μg/mL, and 92.20 ± 0.30% ABTS radical scavenging rate at 1200 μg/mL. Our current research proposes a selective and high-efficiency substitute for the extraction of saponins and might contribute to further DESs application in the recycling of by-products. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The extraction rate of total saponins and total flavonoids from <span class="html-italic">X. sorbifolia</span> husks using different solvents: (<b>a</b>) the extraction rate of total saponins; (<b>b</b>) the extraction rate of total flavonoids. There is a significant difference when * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01 and *** <span class="html-italic">p</span> &lt; 0.001 versus 70% ethanol.</p> Full article ">Figure 2
<p>The field emission scanning electron microscope images of <span class="html-italic">X. sorbifolia</span> powder: (<b>a</b>) <span class="html-italic">X. sorbifolia</span> crude drug powder; (<b>b</b>) <span class="html-italic">X. sorbifolia</span> powder extracted by water; (<b>c</b>) <span class="html-italic">X. sorbifolia</span> powder extracted by ethanol; (<b>d</b>) <span class="html-italic">X. sorbifolia</span> powder extracted by TPMBr-La.</p> Full article ">Figure 3
<p>Comprehensive score of the response surface of the total saponins extraction rate. (<b>a</b>) The interaction between extraction time and temperature; (<b>b</b>) the interaction between liquid–solid ratio and extraction time; (<b>c</b>) the interaction between extraction time and water content; (<b>d</b>) the interaction between liquid–solid ratio and extraction temperature; (<b>e</b>) the interaction between water content and extraction temperature; (<b>f</b>) the interaction between water content and liquid–solid ratio.</p> Full article ">Figure 4
<p>Adsorption and desorption rates of different kinds of macroporous resins.</p> Full article ">Figure 5
<p>D101 static adsorption kinetic curve of macroporous resin (<b>a</b>); intra-particle diffusion (<b>b</b>); pseudo-first-order (<b>c</b>); pseudo-second-order (<b>d</b>).</p> Full article ">Figure 6
<p>Total ion chromatogram (TIC) 200–2000 <span class="html-italic">m</span>/<span class="html-italic">z</span> of the TPMBr-La-extract saponins of <span class="html-italic">X. sorbifolia</span>.</p> Full article ">Figure 7
<p>Chemical structure of the saponins identified from <span class="html-italic">X. sorbifolia</span> husks.</p> Full article ">Figure 8
<p>Determination of the DPPH and ABTS radical scavenging capacity of bioactive saponins recovered from water, ethanol, and TPMBr-La. There is a significant difference when * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p &lt;</span> 0.001 versus ethanol.</p> Full article ">
20 pages, 1781 KiB  
Article
Polyphenolic Compounds from Lespedeza bicolor Protect Neuronal Cells from Oxidative Stress
by Darya V. Tarbeeva, Evgeny A. Pislyagin, Ekaterina S. Menchinskaya, Dmitrii V. Berdyshev, Anatoliy I. Kalinovskiy, Valeria P. Grigorchuk, Natalia P. Mishchenko, Dmitry L. Aminin and Sergey A. Fedoreyev
Antioxidants 2022, 11(4), 709; https://doi.org/10.3390/antiox11040709 - 3 Apr 2022
Cited by 7 | Viewed by 2428
Abstract
Pterocarpans and related polyphenolics are known as promising neuroprotective agents. We used models of rotenone-, paraquat-, and 6-hydroxydopamine-induced neurotoxicity to study the neuroprotective activity of polyphenolic compounds from Lespedeza bicolor and their effects on mitochondrial membrane potential. We isolated 11 polyphenolic compounds: a [...] Read more.
Pterocarpans and related polyphenolics are known as promising neuroprotective agents. We used models of rotenone-, paraquat-, and 6-hydroxydopamine-induced neurotoxicity to study the neuroprotective activity of polyphenolic compounds from Lespedeza bicolor and their effects on mitochondrial membrane potential. We isolated 11 polyphenolic compounds: a novel coumestan lespebicoumestan A (10) and a novel stilbenoid 5’-isoprenylbicoloketon (11) as well as three previously known pterocarpans, two pterocarpens, one coumestan, one stilbenoid, and a dimeric flavonoid. Pterocarpans 3 and 6, stilbenoid 5, and dimeric flavonoid 8 significantly increased the percentage of living cells after treatment with paraquat (PQ), but only pterocarpan 6 slightly decreased the ROS level in PQ-treated cells. Pterocarpan 3 and stilbenoid 5 were shown to effectively increase mitochondrial membrane potential in PQ-treated cells. We showed that pterocarpans 2 and 3, containing a 3’-methyl-3’-isohexenylpyran ring; pterocarpens 4 and 9, with a double bond between C-6a and C-11a; and coumestan 10 significantly increased the percentage of living cells by decreasing ROS levels in 6-OHDA-treated cells, which is in accordance with their rather high activity in DPPH• and FRAP tests. Compounds 9 and 10 effectively increased the percentage of living cells after treatment with rotenone but did not significantly decrease ROS levels. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Structures of polyphenolic compounds isolated from <span class="html-italic">L. bicolor</span> root bark.</p> Full article ">Figure 1 Cont.
<p>Structures of polyphenolic compounds isolated from <span class="html-italic">L. bicolor</span> root bark.</p> Full article ">Figure 2
<p>Experimental and calculated ECD spectra for compounds <b>9</b> (<b>a</b>) and <b>10</b> (<b>b</b>).</p> Full article ">Figure 3
<p>The influence of polyphenolic compounds from <span class="html-italic">L. bicolor</span> on cell viability (<b>a</b>–<b>c</b>), ROS levels (<b>d</b>), and mitochondrial membrane potential (<b>e</b>) in Neuro-2a cells treated with PQ (1 mM). The percentage of living cells treated with various compounds and PQ was measured by MTT assay. Each bar represents the mean ± SEM of three independent replicates. (*), (**), and (***) indicate, respectively, <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.005, and <span class="html-italic">p</span> &lt; 0.001 versus PQ-treated cells. The difference between control and PQ-treated cells was considered significant.</p> Full article ">Figure 3 Cont.
<p>The influence of polyphenolic compounds from <span class="html-italic">L. bicolor</span> on cell viability (<b>a</b>–<b>c</b>), ROS levels (<b>d</b>), and mitochondrial membrane potential (<b>e</b>) in Neuro-2a cells treated with PQ (1 mM). The percentage of living cells treated with various compounds and PQ was measured by MTT assay. Each bar represents the mean ± SEM of three independent replicates. (*), (**), and (***) indicate, respectively, <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.005, and <span class="html-italic">p</span> &lt; 0.001 versus PQ-treated cells. The difference between control and PQ-treated cells was considered significant.</p> Full article ">Figure 4
<p>The influence of polyphenolic compounds from <span class="html-italic">L. bicolor</span> on cell viability (<b>a</b>–<b>c</b>) and ROS levels (<b>d</b>) in Neuro-2a cells treated with 6-OHDA (80 µM). The percentage of living cells treated with compounds and 6-OHDA was measured by MTT assay. Each bar represents the mean ± SEM of three independent replicates. (*), (**), and (***) indicate, respectively, <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.005, and <span class="html-italic">p</span> &lt; 0.001 versus 6-OHDA-treated cells. The difference between control and 6-OHDA-treated cells was considered significant.</p> Full article ">Figure 5
<p>The influence of polyphenolic compounds from <span class="html-italic">L. bicolor</span> on cell viability (<b>a</b>–<b>c</b>) and ROS levels (<b>d</b>) in Neuro-2a cells treated with rotenone (10 µM). The percentage of living cells treated with compounds and rotenone was measured by the MTT assay. Each bar represents the mean ± SEM of three independent replicates. (*), (**), and (***) indicate, respectively, <span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">p</span> &lt; 0.005, and <span class="html-italic">p</span> &lt; 0.001 versus rotenone-treated cells. The difference between control and rotenone-treated cells was considered significant.</p> Full article ">
14 pages, 1696 KiB  
Article
Impact of Quercetin against Salmonella Typhimurium Biofilm Formation on Food–Contact Surfaces and Molecular Mechanism Pattern
by Pantu Kumar Roy, Min Gyu Song and Shin Young Park
Foods 2022, 11(7), 977; https://doi.org/10.3390/foods11070977 - 28 Mar 2022
Cited by 28 | Viewed by 4223
Abstract
Quercetin is an active nutraceutical element that is found in a variety of foods, vegetables, fruits, and other products. Due to its antioxidant properties, quercetin is a flexible functional food that has broad protective effects against a wide range of infectious and degenerative [...] Read more.
Quercetin is an active nutraceutical element that is found in a variety of foods, vegetables, fruits, and other products. Due to its antioxidant properties, quercetin is a flexible functional food that has broad protective effects against a wide range of infectious and degenerative disorders. As a result, research is required on food-contact surfaces (rubber (R) and hand gloves (HG)) that can lead to cross-contamination. In this investigation, the inhibitory effects of quercetin, an antioxidant and antibacterial molecule, were investigated at sub-MIC (125; 1/2, 62.5; 1/4, and 31.25; 1/8 MIC, μg/mL) against Salmonella Typhimurium on surfaces. When quercetin (0–125 μg/mL) was observed on R and HG surfaces, the inhibitory effects were 0.09–2.49 and 0.20–2.43 log CFU/cm2, respectively (p < 0.05). The results were confirmed by field emission scanning electron microscopy (FE-SEM), because quercetin inhibited the biofilms by disturbing cell-to-cell connections and inducing cell lysis, resulting in the loss of normal cell morphology, and the motility (swimming and swarming) was significantly different at 1/4 and 1/2 MIC compared to the control. Quercetin significantly (p < 0.05) suppressed the expression levels of virulence and stress response (rpoS, avrA, and hilA) and quorum-sensing (luxS) genes. Our findings imply that plant-derived quercetin could be used as an antibiofilm agent in the food industry to prevent S. Typhimurium biofilm formation. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Swimming motility assay for <span class="html-italic">Salmonella</span> Typhimurium with various amount of quercetin (μg/mL). Data are expressed as mean ± SEM of three independent replicates. <sup>a–c</sup> Values with different letters are significantly different by Duncan’s multiple-range test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Swarming motility assay for <span class="html-italic">Salmonella</span> Typhimurium with various amount of quercetin (μg/mL). Data are expressed as mean ± SEM of three independent replicates. <sup>a–c</sup> Values with different letters are significantly different by Duncan’s multiple-range test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Inhibition of <span class="html-italic">Salmonella</span> Typhimurium biofilm formation (24 h) on rubber surfaces by various concentrations of quercetin (1/8, 1/4, and 1/2 MIC). Data are expressed as mean ± SEM of three independent replicates. <sup>a–c</sup> Values with different letters are significantly different by Duncan’s multiple-range test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Inhibition of <span class="html-italic">Salmonella</span> Typhimurium biofilm formation (24 h) on hand gloves surfaces by various concentrations of quercetin (μg/mL). Data are expressed as mean ± SEM of three independent replicates. <sup>a–c</sup> Values with different letters are significantly different by Duncan’s multiple-range test (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Representative scanning electron micrographs of <span class="html-italic">Salmonella</span> Typhimurium biofilms formation in the presence of various amounts of quercetin on the hand gloves surfaces. (<b>A</b>) Blank; (<b>B</b>) Control (0% quercetin); (<b>C</b>) 1/8 MIC; (<b>D</b>) 1/2 MIC.</p> Full article ">Figure 6
<p>Relative expression levels of <span class="html-italic">rpoS</span>, <span class="html-italic">avrA</span>, <span class="html-italic">hilA</span>, and <span class="html-italic">luxS</span> genes in <span class="html-italic">Salmonella</span> Typhimurium suspension, supplemented with various amounts of quercetin. <sup>a,b</sup> Different superscript letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) with three independent replicates.</p> Full article ">
14 pages, 3528 KiB  
Article
Ripening-Induced Changes in the Nutraceutical Compounds of Differently Coloured Pepper (Capsicum annuum L.) Breeding Lines
by Zsófia Kovács, Janka Bedő, Bánk Pápai, Andrea Kitti Tóth-Lencsés, Gábor Csilléry, Antal Szőke, Éva Bányai-Stefanovits, Erzsébet Kiss and Anikó Veres
Antioxidants 2022, 11(4), 637; https://doi.org/10.3390/antiox11040637 - 26 Mar 2022
Cited by 8 | Viewed by 2625
Abstract
To date, several research studies addressed the topic of phytochemical analysis of the different coloured pepper berries during ripening, but none discussed it in the case of purple peppers. In this study we examine whether the anthocyanin accumulation of the berries in the [...] Read more.
To date, several research studies addressed the topic of phytochemical analysis of the different coloured pepper berries during ripening, but none discussed it in the case of purple peppers. In this study we examine whether the anthocyanin accumulation of the berries in the early stages of ripening could result in a higher antioxidant capacity due to the elevated amount of polyphenolic compounds. Therefore, enzymatic and non-enzymatic antioxidant capacity was measured in four distinct phenophases of fruit maturity. Furthermore, the expression of structural and regulatory genes of the anthocyanin biosynthetic pathway was also investigated. An overall decreasing trend was observed in the polyphenolic and flavonoid content and antioxidant capacity of the samples towards biological ripeness. Significant changes both in between the genotypes and in between the phenophases were scored, with the genotype being the most affecting factor on the phytonutrients. An extreme purple pepper yielded outstanding results compared to the other genotypes, with its polyphenolic and flavonoid content as well as its antioxidant capacity being the highest in every phenophase studied. Based on our results, besides MYBa (Ca10g11650) two other putative MYBs participate in the regulation of the anthocyanin biosynthetic pathway. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Cross-section of the hypocotyl (<b>a</b>) and berry (<b>b</b>) of ‘Pim. Ney.’.</p> Full article ">Figure 2
<p>Fold expression pattern of the tested genotypes compared to cv. ‘Soroksári’ in 4 phenophases, pseudo-colour bar is showing the level of fold expression on a normalized scale.</p> Full article ">
16 pages, 1442 KiB  
Review
Function, Mechanism, and Application of Plant Melatonin: An Update with a Focus on the Cereal Crop, Barley (Hordeum vulgare L.)
by Xinxing Yang, Jie Chen, Yuan Ma, Minhua Huang, Ting Qiu, Hongwu Bian, Ning Han and Junhui Wang
Antioxidants 2022, 11(4), 634; https://doi.org/10.3390/antiox11040634 - 25 Mar 2022
Cited by 33 | Viewed by 3807
Abstract
Melatonin is a multiple-function molecule that was first identified in animals and later in plants. Plant melatonin regulates versatile processes involved in plant growth and development, including seed germination, root architecture, flowering time, leaf senescence, fruit ripening, and biomass production. Published reviews on [...] Read more.
Melatonin is a multiple-function molecule that was first identified in animals and later in plants. Plant melatonin regulates versatile processes involved in plant growth and development, including seed germination, root architecture, flowering time, leaf senescence, fruit ripening, and biomass production. Published reviews on plant melatonin have been focused on two model plants: (1) Arabidopsis and (2) rice, in which the natural melatonin contents are quite low. Efforts to integrate the function and the mechanism of plant melatonin and to determine how plant melatonin benefits human health are also lacking. Barley is a unique cereal crop used for food, feed, and malt. In this study, a bioinformatics analysis to identify the genes required for barley melatonin biosynthesis was first performed, after which the effects of exogenous melatonin on barley growth and development were reviewed. Three integrated mechanisms of melatonin on plant cells were found: (1) serving as an antioxidant, (2) modulating plant hormone crosstalk, and (3) signaling through a putative plant melatonin receptor. Reliable approaches for characterizing the function of barley melatonin biosynthetic genes and to modulate the melatonin contents in barley grains are discussed. The present paper should be helpful for the improvement of barley production under hostile environments and for the reduction of pesticide and fungicide usage in barley cultivation. This study is also beneficial for the enhancement of the nutritional values and healthcare functions of barley in the food industry. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>Comparison of the melatonin biosynthesis pathways in animals and plants. The yellow arrows indicate the pathway of animals, while the green arrows indicate the pathway of plants. Plant SNATs are rate-limiting enzymes catalyzing two reactions. The light green arrows indicate a minor pathway, since the SNAT enzymes of most plants tend to accommodate 5-Methoxytryptamine as the prevailing substrate, rather than serotonin [<a href="#B21-antioxidants-11-00634" class="html-bibr">21</a>,<a href="#B27-antioxidants-11-00634" class="html-bibr">27</a>]. TDC, tryptophan decarboxylase; TPH, tryptophan hydroxylase; T5H, tryptamine 5-hydroxylase; AADC, aromatic amino acid decarboxylase; COMT, caffeic acid <span class="html-italic">O</span>-methyltransferase; SNAT, serotonin <span class="html-italic">N</span>-acetyltransferase; ASDAC, <span class="html-italic">N</span>-acetylserotonin deacetylase; AANAT, arylalkylamine <span class="html-italic">N</span>-acetyltransferase; ASMT, <span class="html-italic">N</span>-acetyl serotonin methyltransferase; HIOMT, hydroxylindole-<span class="html-italic">O</span>-methyltransferase.</p> Full article ">Figure 2
<p>A phylogenetic tree of tryptamine 5-hydroxylase (T5H) homologs in cereal crops and other plant species. The picture was constructed by the Neighbor-Joining method using the MEGA11 software. Bread wheat is an allohexaploid with the AABBDD genome, and, not surprisingly, bread wheat has three sets of melatonin biosynthesis genes. The phylogenetic profiles agree very well with the evolutionary relationships of these organisms.</p> Full article ">Figure 3
<p>The function and mechanism of melatonin in the regulation of plant growth, development, and abiotic stress tolerance. To regulate seed germination, shoot/root growth, flowering time, and fruit ripening and flavoring, melatonin mainly modulates plant hormone crosstalk. To regulate drought, salinity, cold, heavy metals, UV-B, and nanoplastics tolerance, melatonin serves as an antioxidant in plant cells and induces the biosynthesis of polyamines, proline, and other protectants. The dotted lines indicate that the mechanism is unclear.</p> Full article ">Figure 4
<p>Two mechanisms of melatonin on the regulation of plant disease resistance. (<b>A</b>) Melatonin modulates the salicylic acid (SA) and jasmonic acid (JA) signaling cascades to enhance plant innate immunity against infection by various pathogens. The dotted lines indicate that the detailed processes remain unknown. (<b>B</b>) The PMTR1/CAND2 pathway regulates stomatal closure under pathogen infection. flg22: highly conserved N-terminal of flagellin epitope, recognized by FLS2; FLS2/BAK1: Flagellin Sensing 2/Brassinosteroid Insensitive 1-associated kinase 1; GPA1: G protein α subunit; PMTR1/CAND2: phyto-melatonin receptor; MAPK: mitogen-activated protein kinase.</p> Full article ">
23 pages, 4054 KiB  
Article
Optimization of Ultrasound-Assisted Cellulase Extraction from Nymphaea hybrid Flower and Biological Activities: Antioxidant Activity, Protective Effect against ROS Oxidative Damage in HaCaT Cells and Inhibition of Melanin Production in B16 Cells
by Hui-Min Liu, Sheng-Nan Lei, Wei Tang, Meng-Han Xun, Zhi-Wei Zhao, Ming-Yan Cheng, Xiao-Dan Zhang and Wei Wang
Molecules 2022, 27(6), 1914; https://doi.org/10.3390/molecules27061914 - 16 Mar 2022
Cited by 5 | Viewed by 2893
Abstract
In this study, ultrasonic-assisted cellulase extraction (UCE) was applied to extract flavonoids and polyphenols from the Nymphaea hybrid flower. The extraction conditions were optimized using the response surface method (RSM) coupled with a Box-Behnken design. The crude extract of Nymphaea hybrid (NHE) was [...] Read more.
In this study, ultrasonic-assisted cellulase extraction (UCE) was applied to extract flavonoids and polyphenols from the Nymphaea hybrid flower. The extraction conditions were optimized using the response surface method (RSM) coupled with a Box-Behnken design. The crude extract of Nymphaea hybrid (NHE) was further purified using AB-8 macroporous resins, and the purified extract (NHEP) was characterized by FTIR and HPLC. In vitro activity determination by chemical method showed that NHEP displayed strong free radical scavenging abilities against the DPPH and ABTS radicals, good reduction power, and hyaluronidase inhibition. The cell viability by CCK-8 assays showed that NHEP had no significant cytotoxicity for B16 and HaCaT cells when the concentration was below 100 μg/mL and 120 μg/mL, respectively. NHEP with a concentration of 20–160 μg/mL can more effectively reduce the ROS level in H2O2 damaged HaCaT cells compared with 10 μg/mL of VC. The 40 μg/mL of NHEP had similar activity against intracellular melanin production in the B16 melanoma cells compared with 20 μg/mL Kojic acid. Good activities of antioxidation, whitening and protective effect against H2O2-induced oxidative damage promote the potential for NHEP as a functional raw material in the field of cosmetics and medicine. Full article
(This article belongs to the Topic Antioxidant Activity in Plants, Plant-Derived Bioactive Compounds and Foods)
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<p>The influence of different factors on the content of flavonoids in NHE. (<b>a</b>) Liquid-to-solid ratio, (<b>b</b>) Cellulase amount (wt%), (<b>c</b>) Enzymatic hydrolysis time (min), (<b>d</b>) Ethanol concentration (%). The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 3). *: <span class="html-italic">p</span> &lt; 0.05 compared between groups.</p> Full article ">Figure 2
<p>Response surface 3D and contour plots of different factors to effect the flavonoid content of NHE. (<b>a</b>) Liquid-to-solid ratio and cellulase amount; (<b>b</b>) Liquid-to-solid ratio and ethanol concentration; (<b>c</b>) Cellulase amount and ethanol concentration.</p> Full article ">Figure 2 Cont.
<p>Response surface 3D and contour plots of different factors to effect the flavonoid content of NHE. (<b>a</b>) Liquid-to-solid ratio and cellulase amount; (<b>b</b>) Liquid-to-solid ratio and ethanol concentration; (<b>c</b>) Cellulase amount and ethanol concentration.</p> Full article ">Figure 3
<p>Effects of different factors on static adsorption and desorption of AB-8. (<b>a</b>) Adsorption and desorption curve; (<b>b</b>) Influence of pH on adsorption; (<b>c</b>) Influence of sample concentration on adsorption; (<b>d</b>) Influence of ethanol concentration on desorption. The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 3). *: <span class="html-italic">p</span> &lt; 0.05 compared between groups.</p> Full article ">Figure 4
<p>Effects of different factors on dynamic adsorption and desorption of AB-8. (<b>a</b>) Sample volume, (<b>b</b>) Adsorption flow velocity, (<b>c</b>) Adsorption and (<b>d</b>) Desorption curve. The data are shown as the mean ± S.D. (<span class="html-italic">n</span> = 3). The volume of the resin bed (BV) is 25 mL.</p> Full article ">Figure 5
<p>Activities of the NHE, NHEP and positive control by chemical method. (<b>a</b>) DPPH, (<b>b</b>) ABTS, (<b>c</b>) Reduction power, (<b>d</b>) Hyaluronidase inhibition assay. Dg stands for Dipotassium glycyrrhizinate, and the concentration is 10 μg/mL. The date is shown as the mean ± S.D. (<span class="html-italic">n</span> = 3).</p> Full article ">Figure 6
<p>FTIR-NHEP illustrating structural features.</p> Full article ">Figure 7
<p>HPLC of mixed standards (<b>a</b>) and NHEP (<b>b</b>). 1-gallic acid, 2-corilagin, 3-ellagic acid, 4-rutin, 5-myricetin, 6-quercetin, 7-naringin.</p> Full article ">Figure 8
<p>Cytotoxicity of NHEP on HaCaT cells. (<b>a</b>) cell viability of the HaCaT cells cultured with the NHEP for 24 h, (<b>b</b>) cell viability of the HaCaT cells cultured with the NHEP for 48 h, (<b>c</b>) cellular morphology of the HaCaT cells cultured with the NHEP for 24 h and 48 h (scale bar is 100 μm). VC stands for Vitamin C, and the concentration is 10 μg/mL. Significance: compared with the control group, *** <span class="html-italic">p</span> &lt; 0.001. The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 5).</p> Full article ">Figure 9
<p>Cytotoxicity of H<sub>2</sub>O<sub>2</sub> and NHEP on HaCaT cells. (<b>a</b>) viability of the HaCaT cells cultured with the H<sub>2</sub>O<sub>2</sub>, (<b>b</b>) viability of the HaCaT cells cultured with H<sub>2</sub>O<sub>2</sub> and NHEP. VC stands for Vitamin C, and the concentration is 10 μg/mL. Significance: ### <span class="html-italic">p</span> &lt; 0.001 compared with the control group. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with the H<sub>2</sub>O<sub>2</sub> stimulation group. The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 5).</p> Full article ">Figure 10
<p>Effects of NHEP on the ROS level of HaCaT cells with stimulation of H<sub>2</sub>O<sub>2</sub>: (<b>a</b>) ROS production, (<b>b</b>) fluorescence intensity by detected the DCFH-DA. VC stands for Vitamin C, and the concentration is 10 μg/mL (scale bar is 100 μm). Significance: ### <span class="html-italic">p</span> &lt; 0.001 compared with the control group. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared with the H<sub>2</sub>O<sub>2</sub> stimulation group. The data are shown as the mean ± S.D. (<span class="html-italic">n</span> = 5).</p> Full article ">Figure 11
<p>Cytotoxicity of NHEP on B16 cells: (<b>a</b>) viability of the B16 cells cultured with the NHEP for 24 h, (<b>b</b>) viability of the B16 cells cultured with the NHEP for 48 h, (<b>c</b>) cellular morphology of the B16 cells cultured with the NHEP for 24 h and 48 h (scale bar is 100 μm). Significance: compared with the control group, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.0001. The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 5). KA stands for Kojic acid, and the concentration is 20 μg/mL.</p> Full article ">Figure 12
<p>Effects of the NHEP on B16 cells melanin production. Significance: compared with the control group, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001. The data shows as the mean ± S.D. (<span class="html-italic">n</span> = 5). KA stands for Kojic acid, and the concentration is 20 μg/mL.</p> Full article ">
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