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Search Results (288)

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25 pages, 2000 KiB  
Review
The Role of Adipokines between Genders in the Pathogenesis of Osteoarthritis
by Alessio Economou, Ilenia Mallia, Antonella Fioravanti, Stefano Gentileschi, Francesca Nacci, Silvia Bellando Randone, Gemma Lepri and Serena Guiducci
Int. J. Mol. Sci. 2024, 25(19), 10865; https://doi.org/10.3390/ijms251910865 - 9 Oct 2024
Viewed by 339
Abstract
Osteoarthritis (OA) is a chronic, progressive, degenerative joint disease characterized by joint pain, stiffness, and limited movement. It presents significant intra- and inter-individual variability—in particular, between genders. Recent research has increasingly focused on the role of adipokines—especially leptin, adiponectin, and resistin—in the development [...] Read more.
Osteoarthritis (OA) is a chronic, progressive, degenerative joint disease characterized by joint pain, stiffness, and limited movement. It presents significant intra- and inter-individual variability—in particular, between genders. Recent research has increasingly focused on the role of adipokines—especially leptin, adiponectin, and resistin—in the development of OA. Adipokines, peptide hormones primarily secreted by adipose tissue, are involved in crucial physiological processes related to metabolism and immunity. They can also impact bone and cartilage turnover by interacting with joint cells such as osteoblasts, osteoclasts, chondrocytes, and mesenchymal stem cells, thereby linking inflammation with bone cartilage homeostasis. This review aims to elucidate the structure and functions of various adipokines, their serum and synovial levels, and their association with clinical presentation and radiographic progression in OA patients, with a focus on differences between sexes. A narrative literature review was conducted using three databases specifically analyzing sex differences. OA patients generally show elevated serum and synovial levels of leptin, chemerin, and visfatin, as well as high plasma levels of resistin and visfatin. In contrast, synovial levels of adiponectin and omentin are reduced in OA patients compared to healthy individuals, with an inverse relationship to disease severity, suggesting a potential protective role. Resistin and leptin were positively correlated with pain severity and radiographic progression, while adiponectin’s role in OA remains controversial. Regarding sex differences, male OA patients exhibited higher serum levels of leptin, chemerin, and omentin compared to healthy controls, with a positive correlation to the BMI and estrogen levels, potentially explaining the sexual dimorphism observed in this condition. Studies on visfatin and lipocalin did not reveal significant differences in synovial or serum levels between the sexes. The role of resistin remains controversial. Adipokines influence the joint microenvironment and contribute to the progression of osteoarthritis (OA). However, the precise biological mechanisms are not yet fully understood due to the complex interactions between the metabolic, mechanical, and immune systems. Further research is needed to clarify their roles in OA and to identify targeted therapies for managing this degenerative disease. Full article
(This article belongs to the Special Issue Osteoarthritis Biomarkers, Diagnosis and Treatments)
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<p>The role of the major adipokines in the pathogenesis of osteoarthritis, the main pathways influencing bone turnover and the joint microenvironment. Created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.</p>
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<p>The role of minor adipokines in the pathogenesis of osteoarthritis with their pathways. Created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.</p>
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<p>Adipokines, serum and synovial fluid levels, and gender differences. ↑: increased; ↑ (BMI): increasing related to BMI; ↑ (IL-6): increasing related to IL-6 sinovial fluid levels; *: Controversial result. Studies on the expression of resistin in osteoarthritis (OA) across genders have controversial results. A study by Presle did not find significant differences in the serum resistin levels between women and men [<a href="#B39-ijms-25-10865" class="html-bibr">39</a>]. In contrast, the research by Massengale et al. reported higher serum resistin levels in males (9.4 ng/mL) compared to females (7.3 ng/mL) [<a href="#B40-ijms-25-10865" class="html-bibr">40</a>].</p>
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13 pages, 1837 KiB  
Article
Rosehip Extract Decreases Reactive Oxygen Species Production and Lipid Accumulation in Hypertrophic 3T3-L1 Adipocytes with the Modulation of Inflammatory State
by Katarzyna Kowalska and Anna Olejnik
Nutrients 2024, 16(19), 3269; https://doi.org/10.3390/nu16193269 - 27 Sep 2024
Viewed by 499
Abstract
Background: Rosa canina L. (rosehip) is used worldwide in traditional medicine as a plant with medicinal properties. However, its anti-obesity effects are not fully explained on a transcriptional level. Methods: In the present work, the 3T3-L preadipocytes were utilized to explore the impact [...] Read more.
Background: Rosa canina L. (rosehip) is used worldwide in traditional medicine as a plant with medicinal properties. However, its anti-obesity effects are not fully explained on a transcriptional level. Methods: In the present work, the 3T3-L preadipocytes were utilized to explore the impact of R. canina fruit extract (RCE) on the cellular and molecular pathways involved in adipocyte hypertrophy. Results: Obtained results showed the ability of RCE to reduce lipid overloads in hypertrophic adipocytes associated with the down-regulation of mRNA expressions of adipogenic transcription factors such as PPARγ, C/EBPα, and SREBP-1c as well as genes involved in lipid biosyntheses such as FAS, LPL, and aP2. Moreover, obesity-associated oxidative stress (antioxidant enzyme activities and ROS generation) and inflammation were ameliorated in RCE-treated hypertrophic adipocytes. The mRNA and protein levels of adipokines such as leptin, resistin, and adiponectin were restored to more favorable levels. Conclusions: Rosa canina fruit might be a valuable source of phytochemicals in preventing obesity and obesity-related metabolic complications. Full article
(This article belongs to the Special Issue Effects and Modulatory Mechanisms of Dietary Flavonoids in Obesity)
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<p>Effect of RCE on lipid amounts after Oil Red O staining (<b>a</b>) and lipogenic genes’ expression (<b>b</b>,<b>c</b>) in hypertrophic 3T3-L1 adipocytes. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>a</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>b</sup> <span class="html-italic">p</span> &lt; 0.01, <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001. The images show the hypertrophic adipocytes after 14 days of differentiation (<b>d</b>), Oil Red-stained adipocytes non-treated (<b>e</b>)<b>,</b> and treated with 5 mg/mL RCE (<b>f</b>). The images in (<b>g</b>) show the different steps of the differentiation process, from preadipocytes to hypertrophic adipocytes. Adipocytes were photographed at a magnification of 100×.</p>
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<p>Effect of RCE on lipid amounts after Oil Red O staining (<b>a</b>) and lipogenic genes’ expression (<b>b</b>,<b>c</b>) in hypertrophic 3T3-L1 adipocytes. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>a</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>b</sup> <span class="html-italic">p</span> &lt; 0.01, <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001. The images show the hypertrophic adipocytes after 14 days of differentiation (<b>d</b>), Oil Red-stained adipocytes non-treated (<b>e</b>)<b>,</b> and treated with 5 mg/mL RCE (<b>f</b>). The images in (<b>g</b>) show the different steps of the differentiation process, from preadipocytes to hypertrophic adipocytes. Adipocytes were photographed at a magnification of 100×.</p>
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<p>Effect of rosehip extract (RCE) on the ROS production after NBT staining procedure (<b>a</b>), and pro-oxidant (<b>b</b>) and antioxidant enzymes’ (<b>c</b>–<b>e</b>) mRNA expression in hypertrophic adipocytes. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The effect of RCE on adiponectin (<b>a</b>), leptin (<b>b</b>), and resistin (<b>c</b>) gene expression and adiponectin (<b>e</b>), leptin (<b>f</b>), and resistin (<b>d</b>) protein secretion by hypertrophic adipocytes. Budesonide (BD) was used as a positive control. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>a</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>b</sup> <span class="html-italic">p</span> &lt; 0.01, <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The effect of RCE on adiponectin (<b>a</b>), leptin (<b>b</b>), and resistin (<b>c</b>) gene expression and adiponectin (<b>e</b>), leptin (<b>f</b>), and resistin (<b>d</b>) protein secretion by hypertrophic adipocytes. Budesonide (BD) was used as a positive control. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>a</sup> <span class="html-italic">p</span> &lt; 0.05, <sup>b</sup> <span class="html-italic">p</span> &lt; 0.01, <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The effect of RCE on pro-inflammatory IL-6 and MCP-1 (<b>a</b>) and anti-inflammatory IL-10 (<b>b</b>) gene expression in hypertrophic adipocytes. Budesonide (BD) was used as a positive control. Data are the mean values ± SD (<span class="html-italic">n</span> = 3). <sup>b</sup> <span class="html-italic">p</span> &lt; 0.01, <sup>c</sup> <span class="html-italic">p</span> &lt; 0.001.</p>
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13 pages, 306 KiB  
Article
Mediterranean Diet Prior to Ischemic Stroke and Potential Circulating Mediators of Favorable Outcomes
by María Castañón-Apilánez, Carmen García-Cabo, Cristina Martin-Martin, Belén Prieto, Eva Cernuda-Morollón, Pablo Rodríguez-González, Daniela Pineda-Cevallos, Lorena Benavente, Sergio Calleja and Elena López-Cancio
Nutrients 2024, 16(18), 3218; https://doi.org/10.3390/nu16183218 - 23 Sep 2024
Viewed by 821
Abstract
Background/Objectives. A Mediterranean diet (MD) has been associated with neuroprotective effects. We aimed to assess the MD’s association with stroke prognosis and the potential mediators involved. Methods. Seventy patients with acute anterior circulation ischemic stroke were included. Dietary patterns were evaluated [...] Read more.
Background/Objectives. A Mediterranean diet (MD) has been associated with neuroprotective effects. We aimed to assess the MD’s association with stroke prognosis and the potential mediators involved. Methods. Seventy patients with acute anterior circulation ischemic stroke were included. Dietary patterns were evaluated using the MEDAS scale, a food-frequency questionnaire, and a 24 h recall. Circulating biomarkers including insulin resistance (HOMA index), adipokines (resistin, adiponectin, leptin), choline pathway metabolites (TMAO, betaine, choline), and endothelial progenitor cells (EPCs) were measured. Early neurological improvement (ENI) at 24 h, final infarct volume, and functional outcome at 3 months were assessed. Results. Adherence to MD and olive oil consumption were associated with a lower prevalence of diabetes and atherothrombotic stroke, and with lower levels of fasting glycemia, hemoglobinA1C, insulin resistance, and TMAO levels. Monounsaturated fatty acids and oleic acid consumption correlated with lower resistin levels, while olive oil consumption was significantly associated with EPC mobilization. Multivariate analysis showed that higher MD adherence was independently associated with ENI and good functional prognosis at 3 months. EPC mobilization, lower HOMA levels, and lower resistin levels were associated with ENI, a smaller infarct volume, and good functional outcome. Conclusions. MD was associated with better prognosis after ischemic stroke, potentially mediated by lower insulin resistance, increased EPC mobilization, and lower resistin levels, among other factors. Full article
(This article belongs to the Special Issue Diet, Nutrition and Cardiovascular Health)
15 pages, 1533 KiB  
Article
10-Gingerol Increases Antioxidant Enzymes and Attenuates Lipopolysaccharide-Induced Inflammation by Modulating Adipokines in 3T3-L1 Adipocytes
by María Elizabeth Preciado-Ortiz, Erika Martínez-López, José Pedraza-Chaverri, Omar Noel Medina-Campos, Roberto Rodríguez-Echevarría, Samantha Desireé Reyes-Pérez and Juan José Rivera-Valdés
Antioxidants 2024, 13(9), 1093; https://doi.org/10.3390/antiox13091093 - 7 Sep 2024
Viewed by 1014
Abstract
Background: Obesity increases reactive oxygen species production and alters adipokines levels, resulting in a low-grade chronic inflammation state, which contributes to tissue metabolic dysfunction. 10-gingerol, a phenol present in ginger, has shown potential anti-obesogenic effects in vitro. However, the antioxidant and anti-inflammatory properties [...] Read more.
Background: Obesity increases reactive oxygen species production and alters adipokines levels, resulting in a low-grade chronic inflammation state, which contributes to tissue metabolic dysfunction. 10-gingerol, a phenol present in ginger, has shown potential anti-obesogenic effects in vitro. However, the antioxidant and anti-inflammatory properties of 10-gingerol have not been approached. The aim of this study was to investigate the effects of 10-gingerol on antioxidant enzymes’ expression and adipokine production in 3T3-L1 adipocytes in response to lipopolysaccharide (LPS)-induced inflammation. Methods: 10-gingerol antioxidant capacity was assessed through Oxygen Radical Absorbance Capacity (ORAC) , Ferric Reducing Antioxidant Power (FRAP), and radical scavenging activity of 2,2-diphenyl-2-picrylhydrazyl (DPPH) assays. 3T3-L1 cells were differentiated and stimulated with 100 ng/mL LPSs. Then, 15 µg/mL 10-gingerol was added for 48 h. The mRNA expression and protein abundance of antioxidant enzymes were evaluated by qPCR and Western blot, respectively. Adipokine levels were determined by ELISA. Results: 10-gingerol showed low FRAP and DPPH values but a moderate ORAC value. Moreover, 10-gingerol increased Gpx1 and Sod1 but downregulated Cat expression. Additionally, 10-gingerol significantly increased CAT and GPx1 levels but not SOD-1. Finally, adiponectin and leptin concentrations were increased while resistin and tumor necrosis factor alpha (TNFα) were decreased by 10-gingerol. Conclusions: 10-gingerol presented antioxidant potential by increasing antioxidant enzymes and attenuated LPS-induced inflammation by modulating adipokines in 3T3-L1 adipocytes. Full article
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<p>In vitro study design to determine 10-gingerol effects in 3T3-L1 adipocytes in response to lipopolysaccharide-induced inflammation.</p>
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<p>Percentage of DPPH radical scavenging activity of 10-gingerol. Values plotted represent mean ± SEM. The assays were performed in triplicate. DPPH: 2,2-diphenyl-1-picrylhydrazyl radical.</p>
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<p>Effect of 10-gingerol on mRNA levels (<b>A</b>) <span class="html-italic">Glutathione peroxidase 1</span> (<span class="html-italic">Gpx1)</span>, (<b>B</b>) <span class="html-italic">Superoxide dismutase 1</span> (<span class="html-italic">Sod1</span>), and (<b>C</b>) <span class="html-italic">Catalase</span> (<span class="html-italic">Cat</span>). The relative gene expression of each sample was normalized to <span class="html-italic">Rn18s</span>. Values plotted represent mean ± SEM. The assays were performed in quadruplicate. Differences were analyzed using one-way ANOVA and Tukey’s post hoc. * <span class="html-italic">p</span> &lt; 0.05, **** <span class="html-italic">p</span> &lt; 0.0001. LPS, lipopolysaccharide.</p>
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<p>10-gingerol increases antioxidant enzyme abundance in 3T3-L1 adipocytes. (<b>A</b>) Glutathione peroxidase 1/2 (GPx 1/2), (<b>B</b>) superoxide dismutase 1 (SOD-1), (<b>C</b>) catalase (CAT), and (<b>D</b>) representative Western blot. β-actin was used as a loading control and was run in a different gel. The assays were performed in duplicate. Values plotted represent mean ± SEM. Differences were analyzed using one-way ANOVA and Tukey’s post hoc. * <span class="html-italic">p</span> &lt; 0.05, **** <span class="html-italic">p</span> &lt; 0.0001. LPS, lipopolysaccharide.</p>
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<p>10-gingerol attenuates LPS-induced inflammation by modulating adipokines in 3T3-L1 adipocytes. 10-gingerol increases (<b>A</b>) adiponectin and (<b>B</b>) leptin concentrations, reduces (<b>C</b>) resistin and (<b>D</b>) TNFα concentrations, but does not affect (<b>E</b>) IL-6 levels in a state of LPS-induced inflammation in 3T3-L1 adipocytes. Values plotted represent mean ± SEM. The assays were performed in triplicate. Differences were analyzed using one-way ANOVA and Tukey’s post hoc. * <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, **** <span class="html-italic">p</span> &lt; 0.0001. LPS, lipopolysaccharide.</p>
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23 pages, 754 KiB  
Review
The Impact of the Endocrine and Immunological Function of Adipose Tissue on Reproduction in Women with Obesity
by Katarzyna Mączka, Olga Stasiak, Paulina Przybysz, Monika Grymowicz and Roman Smolarczyk
Int. J. Mol. Sci. 2024, 25(17), 9391; https://doi.org/10.3390/ijms25179391 - 29 Aug 2024
Viewed by 651
Abstract
Obesity, which leads to metabolic dysregulation and body function impairment, emerges as one of the pressing health challenges worldwide. Excessive body fat deposits comprise a dynamic and biologically active organ possessing its own endocrine function. One of the mechanisms underlying the pathophysiology of [...] Read more.
Obesity, which leads to metabolic dysregulation and body function impairment, emerges as one of the pressing health challenges worldwide. Excessive body fat deposits comprise a dynamic and biologically active organ possessing its own endocrine function. One of the mechanisms underlying the pathophysiology of obesity is low-grade systemic inflammation mediated by pro-inflammatory factors such as free fatty acids, lipopolysaccharides, adipokines (including leptin, resistin and visfatin) and cytokines (TNF-α, IL-1β, Il-6), which are secreted by adipose tissue. Together with obesity-induced insulin resistance and hyperandrogenism, the exacerbated immune response has a negative impact on the hypothalamic–pituitary–gonadal axis at all levels and directly affects reproduction. In women, it results in disrupted ovarian function, irregular menstrual cycles and anovulation, contributing to infertility. This review focuses on the abnormal intracellular communication, altered gene expression and signaling pathways activated in obesity, underscoring its multifactorial character and consequences at a molecular level. Extensive presentation of the complex interplay between adipokines, cytokines, immune cells and neurons may serve as a foundation for future studies in search of potential sites for more targeted treatment of reproductive disorders related to obesity. Full article
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<p>The impact of the secretory function of white adipose tissue on the HPG axis.</p>
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14 pages, 347 KiB  
Article
Development and Validation of a Predictive Score for Three-Year Mortality in Acute Ischemic Stroke Patients
by Ioana Cristina Bârsan, Silvina Iluţ, Nicoleta Tohănean, Raluca Maria Pop, Ştefan Cristian Vesa and Lăcrămioara Perju-Dumbravă
Medicina 2024, 60(9), 1413; https://doi.org/10.3390/medicina60091413 - 29 Aug 2024
Viewed by 498
Abstract
Background and Objectives: Acute ischemic stroke (AIS) is a leading cause of death and disability with poor long-term outcomes. Creating a predictive score for long-term mortality in AIS might be important for optimizing treatment strategies. The aim of this study is to [...] Read more.
Background and Objectives: Acute ischemic stroke (AIS) is a leading cause of death and disability with poor long-term outcomes. Creating a predictive score for long-term mortality in AIS might be important for optimizing treatment strategies. The aim of this study is to develop and validate a predictive score for three-year mortality in patients with AIS using several demographic, clinical, laboratory and imaging parameters. Materials and Methods: This study included 244 AIS patients admitted to a tertiary center and followed up for three years. The patients’ data included demographics, clinical features, laboratory tests (including resistin and leptin levels) and imaging parameters. The patients were randomly divided into a predictive group (n = 164) and a validation group (n = 80). Results: Advanced age, a high NIHSS score, low levels of hemoglobin, elevated resistin levels and the presence of carotid plaques were independently associated with three-year mortality. The predictive model incorporated these variables, and it was validated in a separate cohort. Leptin levels did not significantly predict mortality. Conclusions: This study developed and validated a promising predictive score for three-year mortality in patients with AIS. Advanced age, high NIHSS scores, low hemoglobin levels, elevated resistin levels and the presence of carotid plaques were the independent predictors of long-term mortality. Full article
(This article belongs to the Special Issue New Trends in Acute Ischemic Stroke)
11 pages, 528 KiB  
Article
Resistin and In-Hospital Mortality in Patients with Acute Ischemic Stroke: A Prospective Study
by Ioana Cristina Bârsan, Silvina Iluţ, Nicoleta Tohănean, Raluca Pop, Ştefan Cristian Vesa and Lăcrămioara Perju-Dumbravă
J. Clin. Med. 2024, 13(16), 4889; https://doi.org/10.3390/jcm13164889 - 19 Aug 2024
Viewed by 693
Abstract
Background/Objectives: Understanding the prognostic factors of acute ischemic stroke (AIS) is essential for improving patient outcomes. The aim of this study was to establish the predictive role of plasmatic resistin and leptin on short-term mortality in adult patients with a first episode [...] Read more.
Background/Objectives: Understanding the prognostic factors of acute ischemic stroke (AIS) is essential for improving patient outcomes. The aim of this study was to establish the predictive role of plasmatic resistin and leptin on short-term mortality in adult patients with a first episode of AIS. Methods: This study enrolled 277 patients who were consecutively hospitalized for AIS. Demographic data, cardiovascular risk, comorbidities, and laboratory tests were collected. Death was noted if it occurred during hospitalization. Results: Death was recorded in 33 (11.9%) patients. Conducting multivariate analysis, the following variables were independent variables associated with in-hospital mortality: a resistin value of >11 ng/mL (OR 10.81 (95%CI 2.31;50.57), p = 0.002), a lesion volume of >18.8 mL (OR 4.87 (95%CI 1.87;12.67), p = 0.001), a NIHSS score of >7 (OR 5.88 (95%CI 2.01;17.16), p = 0.001), and the presence of IHD (OR 4.33 (95%CI 1.66;11.27), p = 0.003). This study has some limitations: single-center design (which may affect the generalizability of the results) and the potential impact of the COVID-19 pandemic on patient outcomes. Conclusions: This study demonstrated that resistin is a significant predictor of in-hospital mortality in AIS patients. Other established factors, such as a high NIHSS score, large lesion volume, and the presence of IHD, were reaffirmed as important predictors. Full article
(This article belongs to the Special Issue Review Special Issue Series: Recent Advances in Clinical Neurology)
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<p>Flowchart of the patients included in the study.</p>
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11 pages, 1703 KiB  
Article
Study of the Effects of Deuterium-Depleted Water on the Expression of GLUT4 and Insulin Resistance in the Muscle Cell Line C2C12
by Masumi Kondo, Kaichiro Sawada, Yosuke Matsuda, Makiko Abe, Noriyuki Sanechika, Yumi Takanashi, Yoshitaka Mori, Moritsugu Kimura and Masao Toyoda
Biomedicines 2024, 12(8), 1771; https://doi.org/10.3390/biomedicines12081771 - 6 Aug 2024
Viewed by 842
Abstract
Deuterium-depleted water (DDW) is used in the treatment of many diseases, including cancer and diabetes. To detect the effect of DDW on gene expression and activation of the insulin-responsive transporter GLUT4 as a mechanism for improving the pathology of diabetes, we investigated the [...] Read more.
Deuterium-depleted water (DDW) is used in the treatment of many diseases, including cancer and diabetes. To detect the effect of DDW on gene expression and activation of the insulin-responsive transporter GLUT4 as a mechanism for improving the pathology of diabetes, we investigated the GLUT4 expression and glucose uptake at various concentrations of DDW using the myoblast cell line C2C12 differentiated into myotubes. GLUT4 gene expression significantly increased under deuterium depletion, reaching a maximum value at a deuterium concentration of approximately 50 ppm, which was approximately nine times that of natural water with a deuterium concentration of 150 ppm. GLUT4 protein also showed an increase at similar DDW concentrations. The membrane translocation of GLUT4 by insulin stimulation reached a maximum value at a deuterium concentration of approximately 50–75 ppm, which was approximately 2.2 times that in natural water. Accordingly, glucose uptake also increased by up to 2.2 times at a deuterium concentration of approximately 50 ppm. Drug-induced insulin resistance was attenuated, and the glucose uptake was four times higher in the presence of 10 ng/mL TNF-α and three times higher in the presence of 1 μg/mL resistin at a deuterium concentration of approximately 50 ppm relative to natural water. These results suggest that DDW promotes GLUT4 expression and insulin-stimulated activation in muscle cells and reduces insulin resistance, making it an effective treatment for diabetes. Full article
(This article belongs to the Special Issue Diabetes: Comorbidities, Therapeutics and Insights)
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<p>Effect of DDW on the GLUT4 expression in myodifferentiated C2C12 cells. The expression of GLUT4 was examined at the gene and protein levels in the fully differentiated myotubes of C2C12 cells cultured in media adjusted with various concentrations of DDW for 48 h and stimulated with insulin for 30 min before harvest. (<b>a</b>) Detection of the expression of GLUT4 by qRT-PCR. The graphs show the relative values compared with those of the natural water medium (150 ppm). Each sample number was ≥3. * Significant difference compared with the natural water medium (150 ppm), <span class="html-italic">p</span> &lt; 0.05. (<b>b</b>) Diagram of GLUT4 protein expression detected by Western blotting. The arrow indicates the position of the GLUT4 band (54 KDa).</p>
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<p>Effect of DDW on the membrane translocation of GLUT4. Fully differentiated C2C12 myotubes were cultured for 48 h in media containing various concentrations of DDW. After insulin stimulation to induce the membrane translocation of GLUT4, the cells were harvested and membrane proteins were extracted. GLUT4 protein was detected and quantified using Western blotting. (<b>a</b>) Western blotting of the membrane protein fraction; (<b>b</b>) GLUT4 bands were quantified using ImageJ. Values relative to those in cultures without insulin in natural water medium (150 ppm) are shown. The sample size was four. * Significant difference between the presence and absence of insulin in DDW medium at the same concentration (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Glucose uptake in myotube-differentiated C2C12 cells. Fully myotube-differentiated C2C12 cells were cultured in a medium containing various concentrations of DDW for 48 h, after which the uptake of fluorescence-conjugated glucose was observed. TNF-α and resistin were added 24 h before harvesting, whereas insulin and fluorescence-conjugated glucose were added 30 min before harvesting. Scale bar: 100 μm. (<b>a</b>–<b>c</b>) Images of C2C12 glucose uptake. From the left, first and third rows indicate no insulin added; second and fourth rows indicate insulin added. The upper left two images are phase-contrast images of cells cultured in natural water medium (150 ppm) with and without insulin, respectively. The others are fluorescence observations. (<b>a</b>) Cells without induction of insulin resistance, (<b>b</b>) cells treated with 10 ng/mL TNF-α, and (<b>c</b>) cells treated with 10 μg/mL resistin. (<b>d</b>–<b>f</b>) Fluorescence intensity was measured, and the relative values to culture in natural water medium (150 ppm) without insulin were graphed. The sample size was four. * Significant difference (<span class="html-italic">p</span> &lt; 0.05) between the presence (orange bar) and absence (blue bar) of insulin added to DDW medium at each concentration. (<b>d</b>) Cells without the induction of insulin resistance, (<b>e</b>) cells supplemented with 10 ng/mL TNF-α, and (<b>f</b>) cells supplemented with 10 μg/mL resistin.</p>
Full article ">Figure 3 Cont.
<p>Glucose uptake in myotube-differentiated C2C12 cells. Fully myotube-differentiated C2C12 cells were cultured in a medium containing various concentrations of DDW for 48 h, after which the uptake of fluorescence-conjugated glucose was observed. TNF-α and resistin were added 24 h before harvesting, whereas insulin and fluorescence-conjugated glucose were added 30 min before harvesting. Scale bar: 100 μm. (<b>a</b>–<b>c</b>) Images of C2C12 glucose uptake. From the left, first and third rows indicate no insulin added; second and fourth rows indicate insulin added. The upper left two images are phase-contrast images of cells cultured in natural water medium (150 ppm) with and without insulin, respectively. The others are fluorescence observations. (<b>a</b>) Cells without induction of insulin resistance, (<b>b</b>) cells treated with 10 ng/mL TNF-α, and (<b>c</b>) cells treated with 10 μg/mL resistin. (<b>d</b>–<b>f</b>) Fluorescence intensity was measured, and the relative values to culture in natural water medium (150 ppm) without insulin were graphed. The sample size was four. * Significant difference (<span class="html-italic">p</span> &lt; 0.05) between the presence (orange bar) and absence (blue bar) of insulin added to DDW medium at each concentration. (<b>d</b>) Cells without the induction of insulin resistance, (<b>e</b>) cells supplemented with 10 ng/mL TNF-α, and (<b>f</b>) cells supplemented with 10 μg/mL resistin.</p>
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<p>Glucose uptake in myotube-differentiated C2C12 cells. Fully myotube-differentiated C2C12 cells were cultured in a medium containing various concentrations of DDW for 48 h, after which the uptake of fluorescence-conjugated glucose was observed. TNF-α and resistin were added 24 h before harvesting, whereas insulin and fluorescence-conjugated glucose were added 30 min before harvesting. Scale bar: 100 μm. (<b>a</b>–<b>c</b>) Images of C2C12 glucose uptake. From the left, first and third rows indicate no insulin added; second and fourth rows indicate insulin added. The upper left two images are phase-contrast images of cells cultured in natural water medium (150 ppm) with and without insulin, respectively. The others are fluorescence observations. (<b>a</b>) Cells without induction of insulin resistance, (<b>b</b>) cells treated with 10 ng/mL TNF-α, and (<b>c</b>) cells treated with 10 μg/mL resistin. (<b>d</b>–<b>f</b>) Fluorescence intensity was measured, and the relative values to culture in natural water medium (150 ppm) without insulin were graphed. The sample size was four. * Significant difference (<span class="html-italic">p</span> &lt; 0.05) between the presence (orange bar) and absence (blue bar) of insulin added to DDW medium at each concentration. (<b>d</b>) Cells without the induction of insulin resistance, (<b>e</b>) cells supplemented with 10 ng/mL TNF-α, and (<b>f</b>) cells supplemented with 10 μg/mL resistin.</p>
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12 pages, 921 KiB  
Article
Prevention Is Better than Cure—Body Composition and Glycolipid Metabolism after a 24-Week Physical Activity Program without Nutritional Intervention in Healthy Sedentary Women
by Ewa Śliwicka, Natalia Popierz-Rydlewska, Anna Straburzyńska-Lupa, Jivko Nikolov, Łucja Pilaczyńska-Szcześniak and Anna Gogojewicz
Nutrients 2024, 16(15), 2536; https://doi.org/10.3390/nu16152536 - 2 Aug 2024
Cited by 1 | Viewed by 1179
Abstract
Women are generally less active than men; therefore, the search for an attractive form of physical activity that benefits women’s health is underway. This study aimed to investigate the influence of a 24-week physical activity program on body composition and indices of carbohydrates [...] Read more.
Women are generally less active than men; therefore, the search for an attractive form of physical activity that benefits women’s health is underway. This study aimed to investigate the influence of a 24-week physical activity program on body composition and indices of carbohydrates and lipid metabolism in sedentary, healthy women. The study comprised 18 female volunteers (mean age 35.0 ± 5.3 years). Dietary intake was assessed using a standardized seven-day food record. Before entering the program and after completing it, each participant’s body composition and indices of glycolipid metabolism were measured. Insulin resistance indexes were calculated based on the obtained data. After the physical activity program, significant decreases in body mass and composition, BMI, waist circumference, percentage of fat content, and fat mass were found. Moreover, there was a significant decrease in glucose, insulin, triglycerides (TG), and resistin concentrations, as well as in the mean values of HOMA-IR and HOMA-AD. A substantial increase in adiponectin levels was also found. To conclude, the combined endurance–resistance physical activity program had a beneficial effect on body mass and composition and improved carbohydrate and lipid metabolism in normal-weight, healthy women. Therefore, we recommend this activity to sedentary young women to prevent obesity and metabolic disorders. Full article
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<p>Flowchart of the study selection process.</p>
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13 pages, 560 KiB  
Article
Comparative Effects of Gymnema sylvestre and Berberine on Adipokines, Body Composition, and Metabolic Parameters in Obese Patients: A Randomized Study
by Cindy Bandala, Jazmín Carro-Rodríguez, Noemí Cárdenas-Rodríguez, Itzel Peña-Montero, Modesto Gómez-López, Ana Paola Hernández-Roldán, Juan Carlos Huerta-Cruz, Felipe Muñoz-González, Iván Ignacio-Mejía, Brayan Domínguez and Eleazar Lara-Padilla
Nutrients 2024, 16(14), 2284; https://doi.org/10.3390/nu16142284 - 16 Jul 2024
Viewed by 3077
Abstract
Gymnema sylvestre (GS) and berberine (BBR) are natural products that have demonstrated therapeutic potential for the management of obesity and its comorbidities, as effective and safe alternatives to synthetic drugs. Although their anti-obesogenic and antidiabetic properties have been widely studied, comparative research on [...] Read more.
Gymnema sylvestre (GS) and berberine (BBR) are natural products that have demonstrated therapeutic potential for the management of obesity and its comorbidities, as effective and safe alternatives to synthetic drugs. Although their anti-obesogenic and antidiabetic properties have been widely studied, comparative research on their impact on the gene expression of adipokines, such as resistin (Res), omentin (Ome), visfatin (Vis) and apelin (Ap), has not been reported. Methodology: We performed a comparative study in 50 adult Mexican patients with obesity treated with GS or BBR for 3 months. The baseline and final biochemical parameters, body composition, blood pressure, gene expression of Res, Ome, Vis, and Ap, and safety parameters were evaluated. Results: BBR significantly decreased (p < 0.05) body weight, blood pressure and Vis and Ap gene expression and increased Ome, while GS decreased fasting glucose and Res gene expression (p < 0.05). A comparative analysis of the final measurements revealed a lower gene expression of Ap and Vis (p < 0.05) in patients treated with BBR than in those treated with GS. The most frequent adverse effects in both groups were gastrointestinal symptoms, which attenuated during the first month of treatment. Conclusion: In patients with obesity, BBR has a better effect on body composition, blood pressure, and the gene expression of adipokines related to metabolic risk, while GS has a better effect on fasting glucose and adipokines related to insulin resistance, with minimal side effects. Full article
(This article belongs to the Special Issue Natural Products and Health: 2nd Edition)
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<p>Gene expression of resistin (<b>A</b>,<b>B</b>), omentin (<b>C</b>,<b>D</b>), visfatin (<b>E</b>,<b>F</b>), and apelin (<b>G</b>,<b>H</b>) before and after treatment with <span class="html-italic">Gymnema sylvestre</span> and berberine. * Statistical significance.</p>
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22 pages, 24639 KiB  
Article
Adipokines—A Cohort Prospective Study in Children with Severe Burns
by Silviu Constantin Badoiu, Dan Mircea Enescu, Raluca Tatar, Daniela Miricescu, Iulia-Ioana Stanescu-Spinu, Maria Greabu, Anca Magdalena Coricovac, Silvia Elena Badoiu and Viorel Jinga
Int. J. Mol. Sci. 2024, 25(14), 7630; https://doi.org/10.3390/ijms25147630 - 11 Jul 2024
Viewed by 702
Abstract
Burns generate every year an important burden of morbidity, being a major global public health problem through prolonged hospitalization, complications, and increased mortality. This study’s purpose was to evaluate the serum levels of three adipokines and to establish significant correlations with other circulating [...] Read more.
Burns generate every year an important burden of morbidity, being a major global public health problem through prolonged hospitalization, complications, and increased mortality. This study’s purpose was to evaluate the serum levels of three adipokines and to establish significant correlations with other circulating molecules and with some clinical parameters. We evaluated 32 children with severe burns (over 25% total burned surface area—TBSA) at 48 h, day 10, and day 21 post burn, and 21 controls. The serum levels of adiponectin, resistin, leptin, tumor necrosis factor-α (TNF-α), plasminogen activator inhibitor-1 (PAI-1), and C-reactive protein (CRP) (among nine other biochemical parameters) were detected by Multiplex technique. Significant statistical differences were obtained for resistin and leptin compared to the control group, in different moments of measurements. Adiponectin serum levels presented statistically significant correlations with hot liquid mechanism of burn, the Revised Baux score, TBSA, resistin, PAI-1, CRP, TNF-α, and triglycerides (TGLs) serum levels. Resistin serum levels presented statistically significant correlations with adiponectin, CRP, PAI-1, leptin, and TNF-α. Additionally, we found statistically significant correlations between leptin serum levels and length of hospitalization, TNF-α, resistin, adiponectin, and PAI-1 serum levels. In severely burned children, adiponectin, resistin, and leptin specifically correlate with clinical parameters and with proteins involved in the systemic inflammatory response and the hypermetabolic response. Full article
(This article belongs to the Special Issue Molecular and Biochemical Aspects in Burns)
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<p>The box-plot representation for adiponectin (<b>a</b>), resistin (<b>b</b>), and leptin (<b>c</b>) variations in the target group (serum levels expressed in pg/mL). The values of the studied molecules are medians with interquartile ranges. Friedman’s tests with Dunn–Bonferroni post hoc tests were applied for statistical analysis. A significant difference between measurements is marked by unidentical lowercase letters. No statistically significant differences were observed for adiponectin (<span class="html-italic">p</span> = 0.104), nor for leptin (<span class="html-italic">p</span> = 0.097). In the box-plot graphical representation, the ° symbol marks an outlier for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR. The * symbol marks an extreme outlier, for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR.</p>
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<p>Concentrations, in pg/mL, of the three adipokines (adiponectin (<b>a</b>), resistin (<b>b</b>), and leptin (<b>c</b>)) in the study group and the control group, expressed as medians with IQR. Mann–Whitney U tests for statistical analysis. A significant difference between measurements is marked by unidentical lowercase letters. No statistically significant differences were observed for adiponectin, nor for leptin (<span class="html-italic">p</span> &gt; 0.05). In the box-plot graphical representation, the ° symbol marks an outlier for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR. The * symbol marks an extreme outlier, for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR.</p>
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<p>The box-plot graphic illustration for acute-phase inflammation proteins (CRP (<b>a</b>), TNF-α (<b>b</b>), PAI-1 (<b>c</b>)) and for triacylglycerols (Triglycerides) (<b>d</b>) in the burned children group. The values of the studied molecules are medians with interquartile ranges. Concentrations measured in ng/mL for CRP and PAI-1, pg/mL for TNF-α and mg/dl for triglycerides. Friedman’s tests with Dunn–Bonferroni post hoc tests were applied for statistical analysis. For PAI-1, no statistically significant differences between measurements (<span class="html-italic">p</span> = 0.055) were identified. A significant difference between measurements is marked by unidentical lowercase letters. In the box-plot graphical representation, the ° symbol marks an outlier for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR. The * symbol marks an extreme outlier, for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR.</p>
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<p>Correlations between adiponectin value at T1 and resistin value at T1 (<b>a</b>), CRP level at 48 h (<b>b</b>), and PAI-1 levels at T1 (48 h) (<b>c</b>), along with the correlation between resistin levels at T1 and CRP levels at T1 (<b>d</b>).</p>
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<p>Correlations between resistin levels at T1 and PAI-1 levels at T1 (<b>a</b>), leptin value at T1 and TNF-α at T1 (<b>b</b>), and CRP value at T1 and PAI-1 value at T1 (<b>c</b>).</p>
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<p>Hierarchical clustering of adipokines, inflammatory parameters, and triglyceride values at 48 h (T1). Variables were transformed to base10 logarithm values for data normalization. Data show the clustering of Pearson’s correlation coefficients, where a cluster between adiponectin, resistin, PAI-1, and CRP is indicated by the outlined box.</p>
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<p>Correlations between adiponectin levels at T2 and resistin levels at T2 (<b>a</b>), leptin value at T2 (<b>b</b>), TNF-α value at T2 (<b>c</b>), and PAI-1 levels at T2 (<b>d</b>).</p>
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<p>Correlations between resistin levels at T2 and leptin levels at T2 (<b>a</b>), CRP value at T2 (<b>b</b>), TNF-α value at T2 (<b>c</b>), and PAI-1 levels at T2 (<b>d</b>).</p>
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<p>Correlations between leptin levels at T2 and TNF-α levels at T2 (<b>a</b>), PAI-1 value at T2 (<b>b</b>), and the correlation between TNF-α and PAI-1 values at T2 (<b>c</b>).</p>
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<p>Hierarchical clustering of adipokines, inflammatory parameters, and triglyceride values at 10 days (T2). Variables were transformed to base10 logarithm values for data normalization. Data show the clustering of Pearson’s correlation coefficients, where a cluster between adiponectin, resistin, and PAI-1 is indicated by the outlined box.</p>
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<p>Correlations between adiponectin values at T3 (21 days) and resistin levels at T3 (21 days) (<b>a</b>), PAI-1 levels at T3 (21 days) (<b>b</b>), triglycerides value at T3 (<b>c</b>), along with the correlation between resistin and CRP values at T3 (<b>d</b>).</p>
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<p>Correlation between resistin levels at T3 (21 days) and PAI-1 levels at T3 (21 days) (<b>a</b>), along with the correlations between triglycerides value at T3 and CRP value at T3 (<b>b</b>) and PAI-1 values at T3 (<b>c</b>).</p>
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<p>Hierarchical clustering of adipokines, inflammatory parameters, and triglyceride values at 21 days (T3). Variables were transformed to base10 logarithm values for data normalization. Data show the clustering of Pearson’s correlation coefficients, where a cluster between adiponectin, PAI-1, resistin, CRP, and triglycerides is indicated by the outlined box.</p>
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<p>Correlation between TBSA and adiponectin value at T1.</p>
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<p>Correlations between hospitalization period and leptin value at T2 (<b>a</b>) and T3 (<b>b</b>).</p>
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<p>The three adipokines variations in the target group, as related to the mechanism of burn: hot fluid versus flame (concentrations, in pg/mL). Medians with IQR are depicted. Mann–Whitney U tests—statistical analysis. A significant difference between measurements is marked by unidentical lowercase letters. No statistically significant differences were observed for resistin, nor for leptin (<span class="html-italic">p</span> &gt; 0.05). In the box-plot graphical representation, the ° symbol marks an outlier for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR. The * symbol marks an extreme outlier, for a measurement &lt; 1st quartile-1,5IQR or a measurement &gt; 3rd quartile +1,5IQR.</p>
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<p>Correlation between R-Baux Score and adiponectin value at T1.</p>
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11 pages, 3771 KiB  
Brief Report
Lingonberry (Vaccinium vitis-idaea L.) Skin Extract Prevents Weight Gain and Hyperglycemia in High-Fat Diet-Induced Model of Obesity in Mice
by Riitta Ryyti, Mari Hämäläinen, Tiina Tolonen, Marianne Mäki, Mari Jaakkola, Rainer Peltola and Eeva Moilanen
Nutrients 2024, 16(13), 2107; https://doi.org/10.3390/nu16132107 - 2 Jul 2024
Viewed by 1216
Abstract
The percentage of obese people is increasing worldwide, causing versatile health problems. Obesity is connected to diseases such as diabetes and cardiovascular diseases, which are preceded by a state called metabolic syndrome. Diets rich in fruits and vegetables have been reported to decrease [...] Read more.
The percentage of obese people is increasing worldwide, causing versatile health problems. Obesity is connected to diseases such as diabetes and cardiovascular diseases, which are preceded by a state called metabolic syndrome. Diets rich in fruits and vegetables have been reported to decrease the risk of metabolic syndrome and type 2 diabetes. Berries with a high polyphenol content, including lingonberry (Vaccinium vitis-idaea L.), have also been of interest to possibly prevent obesity-induced metabolic disturbances. In the present study, we prepared an extract from the by-product of a lingonberry juice production process (press cake/pomace) and investigated its metabolic effects in the high-fat diet-induced model of obesity in mice. The lingonberry skin extract partly prevented weight and epididymal fat gain as well as a rise in fasting glucose level in high-fat diet-fed mice. The extract also attenuated high-fat diet-induced glucose intolerance as measured by an intraperitoneal glucose tolerance test (IPGTT). The extract had no effect on the levels of cholesterol, triglyceride or the adipokines adiponectin, leptin, or resistin. The results extend previous data on the beneficial metabolic effects of lingonberry. Further research is needed to explore the mechanisms behind these effects and to develop further health-promoting lingonberry applications. Full article
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<p>Weight gain and food consumption. Body weight development during the study (<b>A</b>), and body (<b>B</b>) and epididymal fat (<b>C</b>) weight at the end of the study. The results are presented in grams (g). The cumulative food intake during the study, expressed as energy unit per body weight (kcal/g) (<b>D</b>). Low-fat diet (LF diet, 10% of energy from fat) or high-fat diet (HF diet, 46% of energy from fat) was given for the animals for six weeks. Mice were dosed each weekday with vehicle (LF and HF diet groups) or lingonberry skin extract (HF + LSE diet group) 8.5 µL/g body weight by intragastric administration. Repeated-measures two-way ANOVA with Bonferroni post-test was applied in the statistical analysis (<b>A</b>). The values in (<b>B</b>–<b>D</b>) were analyzed with one-way ANOVA and Bonferroni post-test. Mean values significantly different from the HF diet group are marked with * = <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, a = 0.068, b = 0.069, and c = 0.113. Values are given as the mean + SEM, <span class="html-italic">n</span> = 12 mice per group.</p>
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<p>The fasting blood glucose (<b>A</b>) and insulin (<b>B</b>) levels and the HOMA-IR index (<b>C</b>) of the mice at the end of the study, and an intraperitoneal glucose tolerance test (IPGTT) one week before the end of the experiment (<b>D</b>). Animals received low-fat diet (LF diet, 10% of energy from fat) or high-fat diet (HF diet, 46% of energy from fat) for six weeks. Mice were dosed each weekday with vehicle (LF and HF diet groups) or lingonberry skin extract (HF + LSE diet group) 8.5 µL/g body weight by intragastric administration. In the IPGTT test (<b>D</b>), the animals received glucose (2 g/kg body weight) by intraperitoneal injection and blood glucose was measured at indicated timepoints thereafter. The results are expressed as mmol/L (<b>A</b>,<b>D</b>) or pmol/L (<b>B</b>). In the statistical analysis, one-way ANOVA and Bonferroni post-test was used in (<b>A</b>–<b>C</b>) and repeated-measures two-way ANOVA with Bonferroni post-test in (<b>D</b>). Mean values significantly different from the HF diet group are marked with * = <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. Values are given as the mean + SEM, <span class="html-italic">n</span> = 12 mice per group.</p>
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19 pages, 7120 KiB  
Article
The Effect of KSK-94, a Dual Histamine H3 and Sigma-2 Receptor Ligand, on Adipose Tissue in a Rat Model of Developing Obesity
by Magdalena Kotańska, Monika Zadrożna, Monika Kubacka, Kamil Mika, Katarzyna Szczepańska, Barbara Nowak, Alessio Alesci, Anthea Miller, Eugenia Rita Lauriano and Katarzyna Kieć-Kononowicz
Pharmaceuticals 2024, 17(7), 858; https://doi.org/10.3390/ph17070858 - 1 Jul 2024
Viewed by 689
Abstract
Background: Numerous studies highlight the critical role that neural histamine plays in feeding behavior, which is controlled by central histamine H3 and H1 receptors. This is the fundamental motivation for the increased interest in creating histamine H3 receptor antagonists as [...] Read more.
Background: Numerous studies highlight the critical role that neural histamine plays in feeding behavior, which is controlled by central histamine H3 and H1 receptors. This is the fundamental motivation for the increased interest in creating histamine H3 receptor antagonists as anti-obesity medications. On the other hand, multiple other neurotransmitter systems have been identified as pharmacotherapeutic targets for obesity, including sigma-2 receptor systems. Interestingly, in our previous studies in the rat excessive eating model, we demonstrated a significant reduction in the development of obesity using dual histamine H3/sigma-2 receptor ligands. Moreover, we showed that compound KSK-94 (structural analog of Abbott’s A-331440) reduced the number of calories consumed, and thus acted as an anorectic compound. Therefore, in this study, we extended the previous research and studied the influence of KSK-94 on adipose tissue collected from animals from our previous experiment. Methods: Visceral adipose tissue was collected from four groups of rats (standard diet + vehicle, palatable diet + vehicle, palatable diet + KSK-94, and palatable diet + bupropion/naltrexone) and subjected to biochemical, histopathological, and immunohistochemical studies. Results: The obtained results clearly indicate that compound KSK-94 prevented the hypertrophy and inflammation of visceral adipose tissue, normalized the levels of leptin, resistin and saved the total reduction capacity of adipose tissue, being more effective than bupropion/naltrexon in these aspects. Moreover, KSK-94 may induce browning of visceral white adipose tissue. Conclusion: Our study suggests that dual compounds with a receptor profile like KSK-94, i.e., targeting histamine H3 receptor and, to a lesser extent, sigma-2 receptor, could be attractive therapeutic options for patients at risk of developing obesity or with obesity and some metabolic disorders. However, more studies are required to determine its safety profile and the exact mechanism of action of KSK-94. Full article
(This article belongs to the Special Issue Histamine Receptor Ligands in Medicinal Chemistry)
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<p>The structure of KSK-94 and affinity at the histamine H<sub>3</sub> and sigma-2 receptors. The key interactions between the ligand and these two biological targets, explaining its high activity, have been described in our previous article [<a href="#B32-pharmaceuticals-17-00858" class="html-bibr">32</a>].</p>
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<p>Effects of KSK-94 (10 mg/kg i.p.) or bupropion/naltrexone (20 + 1 mg/kg i.p.) on resistin (<b>a</b>) and leptin (<b>b</b>) concentration in visceral adipose tissue. Data are expressed as means ± 95% CI, (<span class="html-italic">n</span> = 6), * <span class="html-italic">p</span> &lt; 0.05 vs. standard diet + vehicle, ^ <span class="html-italic">p</span> &lt; 0.05, ^^ <span class="html-italic">p</span> &lt; 0.01 vs. palatable diet + vehicle group (one-way ANOVA, post hoc Tukey multiple comparisons test).</p>
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<p>Decrease in adipocyte number in palatable diet + vehicle group (<b>B</b>) compared to both standard diet + vehicle rats (<b>A</b>) and palatable diet + KSK-94 rats (<b>C</b>). The mean number of adipocytes in the adipose tissue of the palatable diet plus bupropion/naltrexone-treated rats (<b>D</b>) was intermediate between the adipocyte number in standard diet + vehicle (<b>A</b>) and palatable diet + vehicle rats (<b>B</b>). Adipose tissue from palatable diet + vehicle rats (<b>B</b>) showed dead or dying adipocyte surrounded by macrophages forming a crown-like structure (asterisk). Masson’s trichrome staining, scale bars 20 μm.</p>
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<p>Numerical density of adipocytes from visceral adipose tissue of the different groups of rats. Data are expressed as means ± SD, *** <span class="html-italic">p</span> &lt; 0.001 vs. standard diet + vehicle, ^ <span class="html-italic">p</span> &lt; 0.05 vs. palatable diet + vehicle group (Kruskal–Wallis test by ranks, post hoc Dunn test).</p>
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<p>Representative histological pictures of adipose tissue sections from studied groups of rats stained with Masson’s trichrome, showing inflammatory cell infiltration (<b>A</b>) and capillary congestion (<b>B</b>). Numerous inflammatory cells were visible only in palatable diet + vehicle rats. In adipose tissue of palatable diet + KSK-94 rats, only slight inflammation was observed involving mostly septa, but most capillaries were heavily congested (in enlarged box). Scale bars 50 μm.</p>
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<p>Inflammatory cell infiltration (<b>a</b>) and capillary congestion (<b>b</b>) in adipose tissue of the different groups of rats. (<b>a</b>). * <span class="html-italic">p</span> &lt; 0.05 standard diet + vehicle versus palatable diet + vehicle. (<b>b</b>). * <span class="html-italic">p</span> &lt; 0.05 palatable diet + KSK-94 versus palatable diet + bupropion/naltrexone and ** <span class="html-italic">p</span> &lt; 0.01 standard diet + vehicle vs. palatable diet + KSK-94. Data are expressed as median + IQR, Kruskal–Wallis test by ranks, and post hoc Dunn test.</p>
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<p>Histological pictures of visceral adipose tissue sections of palatable diet + KSK-94 group showing beige fat cells surrounded by white adipocytes. Masson’s trichrome stain. Scale bars: (<b>A</b>). 50 μm; (<b>B</b>). 20 μm.</p>
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<p>Effects of KSK-94 (10 mg/kg i.p.) and bupropion/naltrexone (20 + 1 mg/kg i.p.) on UCP-1 concentration in visceral adipose tissue. Data are expressed as means ± 95% CI, (<span class="html-italic">n</span> = 6), one-way ANOVA, post hoc Tukey multiple comparisons test.</p>
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<p>Effects of KSK-94 (10 mg/kg i.p.) and bupropion/naltrexone (20 + 1 mg/kg i.p.) on IL-6 (<b>a</b>) and TNF-a (<b>b</b>) and monocyte chemoattractant protein 1 (MCP-1, (<b>c</b>)) concentration in visceral adipose tissue. Data are expressed as means ± 95% CI, (<span class="html-italic">n</span> = 6), * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001 vs. standard diet + vehicle group (one-way ANOVA, post hoc Tukey multiple comparisons test).</p>
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<p>Immunoexpression of IL-6 was detected in some inflammatory cells (probably in macrophages, arrows, and enlarged box in (<b>C</b>) occurring more frequently in adipose tissue of palatable diet + vehicle rats (<b>B</b>). Only a few cells expressing IL-6 were observed in the adipose tissue of palatable diet+KSK-94 (<b>C</b>) and palatable diet + bupropion/naltrexone rats (<b>D</b>). There were no IL-6-positive cells in the adipose tissue of standard diet + vehicle rats (<b>A</b>). Immunohistochemical reaction counterstain with hematoxylin. Scale bars 50 μm.</p>
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<p>Effects of KSK-94 (10 mg/kg i.p.) and bupropion/naltrexone (20 +1 mg/kg i.p.) on MDA (<b>a</b>) and FRAP (<b>b</b>) concentration in visceral adipose tissue. Data are expressed as means ± 95% CI, (<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 vs. standard diet + vehicle, ^^ <span class="html-italic">p</span> &lt; 0.01 vs. palatable diet + vehicle group (one-way ANOVA, post hoc Tuckey multiple comparisons test).</p>
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<p>Timeline of the experiment.</p>
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16 pages, 9140 KiB  
Article
Supplemental Psyllium Fiber Increases Antimicrobial Proteins via the Tuft Cell-ILC2 Circuit and Type II Immune Response in the Mouse Small Intestine
by Yoshiki Ishii, Taiyo Matsunaga, Tomoki Yasui, Dina Mustika Rini, Ryo Inoue, Yoshinari Yamamoto and Takuya Suzuki
Nutraceuticals 2024, 4(2), 307-322; https://doi.org/10.3390/nutraceuticals4020019 - 13 Jun 2024
Viewed by 3491
Abstract
Dietary fibers regulate intestinal barrier function; however, the precise mechanisms remain unclear. This study investigated the effects of psyllium fibers on antimicrobial protein expression, focusing on the type II immunity and tuft cell-group 2 innate lymphoid cell (ILC2) circuit in the small intestine [...] Read more.
Dietary fibers regulate intestinal barrier function; however, the precise mechanisms remain unclear. This study investigated the effects of psyllium fibers on antimicrobial protein expression, focusing on the type II immunity and tuft cell-group 2 innate lymphoid cell (ILC2) circuit in the small intestine of the mouse. Supplemental psyllium fiber upregulated antimicrobial proteins, such as small proline-rich protein 2A (SPRR2A) and resistin-like beta (RELMβ), in mouse small intestine, evidently affecting cecal microbiota composition. The psyllium fibers also increased the RNA and protein expression of molecules related to ILC2 and tuft cells, such as IL-13, IL-25, DCLK1, Gfi-1b, SH2 domain-containing protein 3C, and Spi-B. In addition, ILC2 inhibitor (disulfiram) and bitter taste receptor blocker administration reduced psyllium-induced SPRR2A and RELMβ expression. Collectively, psyllium supplementation upregulates antimicrobial proteins such as SPRR2A and RELMß via the type II immune response and tuft cell-ILC2 circuit in the mouse small intestine. Full article
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Figure 1

Figure 1
<p>Psyllium supplementation increases antimicrobial protein expression in mouse small intestine. The mice were fed the control and a 10% psyllium diet for 5 d, and jejunal samples were collected. Total RNA from these samples was subjected to RNA sequencing and qRT-PCR. Data (transcripts per million, TPM) were log 2 transformed after the addition of 1 for TPM, and the MA plot is shown (<b>A</b>). “Average” represents the mean values of the control and 10% psyllium groups. “Minus” represents the differences between the values of the control and 10% psyllium groups. Each dot indicates a specific gene, and red and blue dots indicate genes &gt;2-fold higher or lower in the 10% psyllium group than those in the control group, respectively. The heatmap shows the Z-scores of the genes encoding antimicrobial proteins (<b>B</b>). <span class="html-italic">Srpp2a</span> and <span class="html-italic">Retnlb</span> mRNA expression levels were determined using qRT-PCR (<b>C</b>,<b>D</b>). The protein expression of the levels of SPRR2A and RELMβ were examined using immunoblot (<b>E</b>,<b>F</b>) and immunofluorescence (<b>G</b>,<b>H</b>) analyses. Results are shown as the mean ± SEM; n = 8. Statistical analysis was performed using an unpaired Student’s <span class="html-italic">t</span>-test, * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 1</span>.</p>
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<p>Psyllium supplementation alters the microbiome profile in the mouse cecum. The mice were fed the control and a 10% psyllium diet for 5 d, and the fresh cecal samples were subjected to 16S rRNA sequencing. The number of observed species (<b>A</b>), Chao1 index (<b>B</b>), and Shannon index (<b>C</b>) of the cecal microbiota were calculated. Principal coordinate analysis (PCoA) plots of intestinal microbiota were generated based on weighted UniFrac (<b>D</b>), unweighted UniFrac (<b>E</b>), and Bray-Curtis metrics (<b>F</b>) using QIIME. (<b>G</b>) The relative abundance of cecal microbiota at the phylum level. Genera with statistically significant differences observed between groups are shown (<b>H</b>,<b>I</b>). The genera representing relative abundances &gt;5% (<b>H</b>) and 1% (<b>I</b>) in either group are shown. Results are shown as the mean ± SEM, n = 8. Statistical analysis was performed using an unpaired Student’s <span class="html-italic">t</span>-test, * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 1</span>.</p>
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<p>Psyllium supplementation upregulates antimicrobial proteins, SPRR2A, and RELMβ in mouse jejunum and ileum. Mice were fed diets with and without psyllium fiber (2.5, 5, and 10%) for 5 d. Jejunal and ileal samples were subjected to qRT−PCR and immunoblotting. The mRNA expression levels of <span class="html-italic">Sprr2a</span> (<b>A</b>,<b>E</b>) and <span class="html-italic">Retnlb</span> (<b>C</b>,<b>G</b>) levels were determined in the mouse jejunum (<b>A</b>,<b>C</b>) and ileum (<b>E</b>,<b>G</b>) using qRT−PCR. The protein expression levels of SPRR2A (<b>B</b>,<b>F</b>) and RELMß (<b>D</b>,<b>H</b>) were determined in the mouse jejunum (<b>B</b>,<b>D</b>) and ileum (<b>F</b>,<b>H</b>) by immunoblot analysis. Results are shown as the mean ± SEM; n = 6. Statistical analysis was performed using the Tukey−Kramer post hoc test and Steel−Dwass test, * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 2</span>.</p>
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<p>Psyllium supplementation promotes tuft cell proliferation and the ILC2 immune response. The mice were fed diets with and without psyllium fiber (2.5, 5, 10%) for 5 d, and the jejunal samples were subjected to qRT-PCR, immunoblot, and immunofluorescence analyses. The mRNA expression levels of <span class="html-italic">Pou2f3</span> (<b>A</b>), <span class="html-italic">Dclk1</span> (<b>B</b>), <span class="html-italic">Gfi1b</span> (<b>C</b>), <span class="html-italic">Sh2d6</span> (<b>D</b>), <span class="html-italic">Tppp3</span> (<b>E</b>), <span class="html-italic">Spib</span> (<b>F</b>), <span class="html-italic">Il25</span> (<b>H</b>), <span class="html-italic">Il13</span> (<b>I</b>), and <span class="html-italic">Il4</span> (<b>J</b>) were determined using qRT−PCR. A tuft cell marker, DCLK1, in the mouse jejunum was visualized via immunofluorescence (scale bar: 200 µm), and the number of DCLK1<sup>+</sup> tuft cells in a villus was quantified (<b>G</b>). pSTAT6 protein expression in mouse jejunum was determined using immunoblot analysis (<b>K</b>). Results are shown as the mean ± SEM; n = 6. Statistical analyses were performed using the Tukey−Kramer post hoc test and unpaired Student’s <span class="html-italic">t</span>−test; * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 2</span>.</p>
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<p>ILC2 is involved in psyllium-induced antimicrobial protein expression. The mice were fed control and 5% psyllium diets for 5 d. An ILC2 inhibitor, disulfiram, was administered orally at 1.2 and 2.4 mg/g body weight 2 d before the start of the test diets. The jejunal samples were subjected to qRT−PCR, immunoblot, and immunofluorescence analyses. The mRNA expression levels of <span class="html-italic">Sprr2a</span> (<b>A</b>), <span class="html-italic">Retnlb</span> (<b>D</b>), <span class="html-italic">Pou2f3</span> (<b>G</b>), <span class="html-italic">Il25</span> (<b>H</b>), and <span class="html-italic">Il13</span> (<b>J</b>) in the mouse jejunum were determined using qRT-PCR analysis. The protein expression levels of SPRR2A (<b>B</b>), RELMβ (<b>E</b>), and pSTAT6 (<b>K</b>) were determined by immunoblot analysis. The expression of SPRR2A (<b>C</b>), RELMβ (<b>F</b>), DCLK1 (<b>I</b>), and pSTAT6 (<b>L</b>) was visualized using immunofluorescent analysis (scale bar: 200 µm). Results are shown as the mean ± SEM; n = 7. Statistical analysis was performed using the Tukey−Kramer post hoc test, * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 4</span>.</p>
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<p>Bitter taste receptors are involved in the psyllium-induced antimicrobial protein expression. The mice were fed control and 5% psyllium diets with and without bitter taste receptor blocker cocktail (0.25 and 0.5%) for 5 d. The jejunal samples were subjected to qRT−PCR, immunoblot, and immunofluorescence analyses. The mRNA expression levels of <span class="html-italic">Sprr2a</span> (<b>A</b>), <span class="html-italic">Retnlb</span> (<b>D</b>), <span class="html-italic">Pou2f3</span> (<b>G</b>), <span class="html-italic">Il25</span> (<b>H</b>), and <span class="html-italic">Il13</span> (<b>J</b>) in the mouse jejunum were determined using qRT-PCR analysis. The protein expression levels of SPRR2A (<b>B</b>), RELMβ (<b>E</b>), and pSTAT6 (<b>K</b>) were determined using immunoblot analysis. The expression of SPRR2A (<b>C</b>), RELMβ (<b>F</b>), DCLK1 (<b>I</b>), and pSTAT6 (<b>L</b>) was visualized using immunofluorescent analysis (Scale bar: 200 µm). Results are shown as the mean ± SEM; n = 7. Statistical analysis was performed using the Tukey−Kramer post hoc test, * <span class="html-italic">p</span> &lt; 0.05, related to <span class="html-italic">experiment 5</span>.</p>
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23 pages, 1854 KiB  
Article
Effects of Aging on Secreted Adipocytokines in Visceral Fat of Female C3H/HeJ Mice Consuming a Long-Term High-Fat Diet
by Noshin Mubtasim, Benjamin Barr, Caleb Boren and Lauren Gollahon
Dietetics 2024, 3(2), 191-213; https://doi.org/10.3390/dietetics3020016 - 13 Jun 2024
Viewed by 812
Abstract
The excess consumption of a high-fat diet has been identified as one of the factors contributing to obesity. Women are at higher risk of adult obesity than men, predisposing them to a different set of detrimental disease conditions. Furthermore, aging studies show that [...] Read more.
The excess consumption of a high-fat diet has been identified as one of the factors contributing to obesity. Women are at higher risk of adult obesity than men, predisposing them to a different set of detrimental disease conditions. Furthermore, aging studies show that physiological decline also has a serious impact on changes in the endocrine properties of white adipose tissue. However, there is still relatively little known about the factors associated with obesity and aging and their compounding impacts on women’s health. To investigate changes in adipocytokine secretion profiles, obesity was induced in female C3H/HeJ mice through the long-term consumption of a high-fat diet. Weight gain measurements and the Echo MRI analysis of fat composition showed that increases were due solely to the high fat content in the diet. Adipocytokine secretions were analyzed in media conditioned from harvested visceral fat tissue that was organ-cultured ex vivo. Adipocytokine analysis performed across diets and ages showed that there were significant increases in Adiponectin and Leptin secretion in high-fat diets, accelerating increases in Resistin secretion in high-fat diets. Aging induced the increased secretion of Lipcalin-2, Pentraxin-3, Serpin E1, MCP-1, and ICAM-1, regardless of diet. Furthermore, the comparisons of organoid-cultured protein secretions and flash-frozen tissue samples differed greatly, suggesting the WAT organoid cultures may yield information that is more reflective of in situ conditions. Taken together, our results show that high-fat diets and aging in C3H/HeJ female mice significantly impact secretions from adipose tissue, which may contribute to women’s health issues. Full article
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Figure 1
<p>Schematic representation of mouse adipokine array and coordinates for antibodies against 38 commonly occurring adipocytokines.</p>
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<p>Adipokine array showing dot blot results of different adipocytokines on nitrocellulose membrane. Dots are adipocytokine signals detected after treatment with a visceral, fat-derived, conditioned medium harvested from C3H/HeJ female mice fed on control or high-fat diets, both for Month 12 and Month 18. (<b>A</b>) The negative control is M199 media without FBS, used for conditioning cells. (<b>B</b>) Organ-cultured visceral fat harvested from control-diet-fed mice at Month 12. (<b>C</b>) Organ-cultured visceral fat harvested from high-fat-diet-fed mice at Month 12. (<b>D</b>) Organ-cultured visceral fat harvested from control fat-diet-fed mice at Month 18. (<b>E</b>) Organ-cultured visceral fat harvested from high-fat-diet-fed mice at Month 18.</p>
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<p>Changes in adipocytokine secretions, (<b>A</b>) Adiponectin, (<b>B</b>) Leptin, (<b>C</b>) MCP-1, (<b>D</b>) Resistin, (<b>E</b>) TIMP-1, and (<b>F</b>) VEGF in the Ex-WAT-CM harvested from control and high-fat-diet-fed female C3H/HeJ mice at Month 12 and Month 18. Each data point represents the mean value of the secretome from 2 technical replicates for 3 independent experiments. Adipocytokine secretion changes over time between the control and high-fat diet; this was determined using two-way ANOVA, followed by Tukey’s post hoc test for multiple comparisons. A <span class="html-italic">p</span> value ≤ 0.05 was considered statistically significant and identified by one asterisk (*); two asterisks (**) indicate <span class="html-italic">p</span> ≤ 0.01; three asterisks (***) indicate <span class="html-italic">p</span> ≤ 0.001; and four asterisks (****) indicate <span class="html-italic">p</span> ≤ 0.0001. <span class="html-italic">p</span>-value &gt; 0.05 was considered statistically nonsignificant and not marked on the graphs. Note: As ExV-WAT-CM samples are conditioned media collected from ex vivo visceral adipose tissue, the changes in adipocytokine secretions between the conditioned medium are designated as arbitrary units for the <span class="html-italic">y</span>-axis, following the manufacturer’s kit instructions.</p>
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<p>Changes in adipocytokine secretions, (<b>A</b>) IL-6, (<b>B</b>) ICAM-1, (<b>C</b>) Lipocalin-2, (<b>D</b>) Serpin E1, (<b>E</b>) Pentraxin-3, and (<b>F</b>) M-CSF in the ExV-WAT-CM harvested from control and high-fat-diet-fed mice female C3H/HeJ mice at Month 12 and Month 18. Each data point represents the mean value of the secretome from 2 technical replicates for 3 independent experiments. Adipocytokine secretion changes over time, between control and high-fat diets, were measured using two-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. A <span class="html-italic">p</span> value <span class="html-italic">p</span> ≤ 0.05 was considered statistically significant and identified by one asterisk (*), two asterisks (**) indicate <span class="html-italic">p</span> ≤ 0.01, and three asterisks (***) indicate <span class="html-italic">p</span> ≤ 0.001. Note: As ExV-WAT-CM samples are conditioned media collected from ex vivo visceral adipose tissue, the changes in adipocytokine secretions between the conditioned medium as designated as arbitrary units for the <span class="html-italic">y</span>-axis, following the manufacturer’s kit instruction.</p>
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<p>Graphical representation of observable changes in the expression of adipocytokine proteins extracted from visceral fat flash-frozen tissue (FF-WAT) and harvested from control and high-fat-diet-fed mice at Month 12 and Month 18. (<b>A</b>) Adiponectin, (<b>B</b>) Leptin, (<b>C</b>) MCP-1, (<b>D</b>) Resistin, (<b>E</b>) TIMP-1, and (<b>F</b>) VEGF in the FF-WAT tissue harvested from control and high-fat-diet-fed female C3H/HeJ mice at Month 12 and Month 18. Each data point represents the mean value of the protein analyzed from 3 technical replicates for 3 independent experiments. Adipocytokine protein changes over time between control and high-fat diets were determined using two-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. A <span class="html-italic">p</span> value ≤ 0.05 was considered statistically significant and identified by one asterisk (*), and two asterisks (**) indicate <span class="html-italic">p</span> ≤ 0.01. The changes in adipocytokine protein levels have been designated as arbitrary units for the <span class="html-italic">y</span>-axis following the manufacturer’s kit instruction. Note that the scale breaks in the <span class="html-italic">y</span>-axis units based on protein expression levels.</p>
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