Pueraria lobata–Prunus mume Complex Alleviates Alcoholic Liver Disease by Regulating Lipid Metabolism and Inhibiting Inflammation: A Transcriptome and Gut Microbiota Analysis
<p>Component analysis of PPC and liver transcriptomics results of mice with ALD. (<b>A</b>) UPLC-MS/MS results of PPC. Black, red, and green lines represent for the UPLC (PDA detector), positive ion mode, and negative ion mode results, respectively. (<b>B</b>) Volcano map to screen for gene expression related to ALD using the screening criteria of |log<sub>2</sub> (fold change)| > 1.5 and <span class="html-italic">p</span> < 0.05. The blue inverted and red positive triangle represent for the downregulated and upregulated genes. (<b>C</b>,<b>D</b>) GO (BP, MF, CC) and KEGG results for the selected genes related to ALD. GO annotations related to lipid metabolism are circled in red.</p> "> Figure 2
<p>16S rRNA results of mice with ALD with/without PPC administration (<span class="html-italic">n</span> = 4). (<b>A</b>) Bar chart of species composition at the phylum level. (<b>B</b>) Differences in alpha-diversity (ACE and Shannon indices). (<b>C</b>) Petal diagram with common unique features. (<b>D</b>) ANOSIM based on the weighted Unifrac algorithm (R = 0.425, <span class="html-italic">p</span> = 0.001). (<b>E</b>) NMDS analysis based on the weighted Unifrac algorithm (Stress = 0.1908). (<b>F</b>) Cladogram of LEfSe. o, order; f, family; g, genus; s, species. Con, control group; Mod, Baijiu treatment group; Pos, silybin group; Low, low PPC dosing group; Med, medium PPC dosing group; Hig, high PPC dosing group.</p> "> Figure 3
<p>KEGG results of 16S rRNA in mice with ALD with/without PPC administration. Mod, Baijiu treatment; Hig, high dose of PPC. An annotation related to lipid metabolism is circled in red. 7.72e-2 = 0.0772, the same representation for the remaining data.</p> "> Figure 4
<p>(<b>A</b>) Weight gain, (<b>B</b>) liver index, and (<b>C</b>) serum biochemical indicators of liver function (AST, ALT, ALP, TC, TG, LDL-C) in mice with ALD in the different groups (<span class="html-italic">n</span> = 10). ELISA results were displayed as enzyme activity concentration (U/L) or molar concentration (mmol/L). Statistical comparisons were made between the control group and the Baijiu group, as well as between the Baijiu group and the treatment group. Data were expressed as the mean ± standard deviation. <span class="html-italic">p</span> ≤ 0.05, 0.01, 0.001 indicates for difference, significant difference, and extremely significant difference in results, respectively. Con, control group; Mod, Baijiu treatment group; Pos, silymarin group; Low, low-dose PPC group; Med, medium-dose PPC group; Hig, high-dose PPC group.</p> "> Figure 5
<p>H&E (HE) and oil red O (ORO) staining of liver tissue. In the histological images, the magnification of the microscope is 10× and 40×, and the length of the scale bar is 100 μm and 20 μm. Black arrow, location of lymphocyte infiltration, red arrow, degenerated and necrotic liver cells. Con, control group; Mod, Baijiu treatment group; Pos, silymarin group; Low, low-dose PPC group; Med, medium-dose PPC group; Hig, high-dose PPC group.</p> "> Figure 6
<p>PPC alleviates inflammation in mice with ALD through antioxidant activity and the restoration of intestinal barrier integrity (<span class="html-italic">n</span> = 10). (<b>A</b>,<b>B</b>) ELISA results of serum antioxidant stress and inflammatory markers (MDA, CAT, GSH, SOD, IL-6, IL-1β and TNF-α), displayed as mass (pg), molar (μmol, nmol) or enzyme activity units (U) of the detection marker per volume units (mL) or mgprot. (<b>C</b>–<b>E</b>) WB results of NF-κB pathway (TLR4, CD14, IκBα, and P65) and intestinal proteins (ZO-1, occludin and claudin-4). Statistical comparisons were made between the control group and the Baijiu model group, as well as between the Baijiu model group and the treatment group. Data were expressed as the mean ± standard deviation. <span class="html-italic">p</span> ≤ 0.05, 0.01, 0.001 indicates for difference, significant difference, and extremely significant difference in results, respectively. Con, control group; Mod, Baijiu treatment group; Pos, silymarin group; Low, low-dose PPC group; Med, medium-dose PPC group; Hig, high-dose PPC group.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Preparation and Liquid Chromatography–Mass Spectrometry (LC-MS)
2.2. Animals and Model Establishment
2.3. Transcriptome and 16S rRNA Analysis
2.4. Serum Biochemical Analysis
2.5. Tissue Staining and Immunohistochemistry
2.6. Western Blotting
2.7. Statistical Analysis
3. Results and Discussion
3.1. Component Analysis of PPC and Liver Transcriptomics Results in Mice with ALD
3.2. 16S rRNA Results of Mice with ALD with/without PPC Administration
3.3. Validation of Liver Transcriptomics and Gut Microbiota Results
3.4. Discussion
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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No | Retention Time (min) | Compound Name | Molecular Weight | Adduct Ion | Molecular Formula | MS/MS Fragments |
---|---|---|---|---|---|---|
1 | 12.90 | 3′-Hydroxypuerarin | 431.0995 | [M−H]− | C21H20O10 | 431.0991, 311.0566, 283.0616 |
2 | 14.99 | Puerarin | 415.1047 | [M−H]− | C21H20O9 | 415.1043, 295.0615, 267.0677 |
3 | 15.80 | 3′-Methoxypuerarin | 445.1153 | [M−H]− | C22H22O10 | 445.1148, 325.0723, 282.0539 |
4 | 16.77 | Puerarin apioside | 547.1473 | [M−H]− | C26H28O13 | 547.1467, 295.0616, 267.0667 |
5 | 17.41 | Daidzin | 417.1183 | [M+H]+ | C21H20O9 | 297.0748, 255.0656, 199.0754 |
6 | 25.07 | Daidzein | 253.0512 | [M−H]− | C15H10O4 | 253.0509, 209.0609, 135.0078 |
7 | 25.57 | Glycitein | 283.0621 | [M−H]+ | C16H12O5 | 283.0616, 268.0381, 211.0394 |
8 | 29.02 | Luteolin | 285.0414 | [M−H]− | C15H10O6 | 285.0409, 255.0300, 151.0031 |
9 | 36.51 | Ursolic acid | 455.3546 | [M−H]− | C30H48O3 | 455.3540, 407.3370, 238.0641 |
10 | 37.43 | Palmitic acid | 255.2334 | [M−H]− | C16H32O2 | 255.2332, 233.1788, 193.8156 |
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Gao, R.; Huang, Q.; Zeng, Y.; Chen, D.; Jia, Z.; Han, B.; Huang, X.; Wang, Q.; Hu, X.; Liao, M.; et al. Pueraria lobata–Prunus mume Complex Alleviates Alcoholic Liver Disease by Regulating Lipid Metabolism and Inhibiting Inflammation: A Transcriptome and Gut Microbiota Analysis. Foods 2024, 13, 2431. https://doi.org/10.3390/foods13152431
Gao R, Huang Q, Zeng Y, Chen D, Jia Z, Han B, Huang X, Wang Q, Hu X, Liao M, et al. Pueraria lobata–Prunus mume Complex Alleviates Alcoholic Liver Disease by Regulating Lipid Metabolism and Inhibiting Inflammation: A Transcriptome and Gut Microbiota Analysis. Foods. 2024; 13(15):2431. https://doi.org/10.3390/foods13152431
Chicago/Turabian StyleGao, Ruixi, Qi Huang, Yanfeng Zeng, Dandan Chen, Ziming Jia, Bingchen Han, Xianju Huang, Qiang Wang, Xin Hu, Maochuan Liao, and et al. 2024. "Pueraria lobata–Prunus mume Complex Alleviates Alcoholic Liver Disease by Regulating Lipid Metabolism and Inhibiting Inflammation: A Transcriptome and Gut Microbiota Analysis" Foods 13, no. 15: 2431. https://doi.org/10.3390/foods13152431