Corilagin prevents non-alcoholic fatty liver disease via improving lipid metabolism and glucose homeostasis in high fat diet-fed mice

Figures

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  FIGURE 1. Cori reduced the high NAFLD activity score (NAS) and inhibited liver damage in HFD-fed mice. (A) H&E staining of livers in indicated groups. As shown in high magnification image of HFD group, arrowhead (yellow color) indicates steatosis; arrow (blue color) indicates inflammatory cells infiltration; asterisk (red color) indicates a balloon-like change; cv indicates central vein. Bar = 50 μm. Liver gross morphology of indicated group was represented in the upper right corner. (B) Hepatic steatosis score, (C) inflammation score, (D) ballooning score, and (E) NAFLD activity score (NAS) were quantified from indicated groups. (F) Serum ALT and (G) AST levels were measured by biochemical analysis. (H) Liver weights of indicated groups. (I) Liver index (the ratio of liver weight to body weight) in indicated groups. NCD, normal chow diet; HFD, high fat diet; HCR-L, high fat diet plus treatment with low dose of Cori (5 mg/kg/2 days); HCR-H, high fat diet plus treatment with high dose of Cori (10 mg/kg/2 days). Values were means ± SD, and for statistical analysis, one-way ANOVA were performed between indicated groups.

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  FIGURE 2. Cori alleviated hepatic and plasma lipid concentrations in HFD-fed mice. (A) Oil Red O staining of liver in indicated groups. Bar = 50 μm. (B) TG and (C) TC levels in liver were detected. (D) Plasm TG, (E) TC, (F) HDL-c, (G) LDH-c were tested. (H) FABP4 in plasma was measured. (I) FFA in plasma was examined. Values were means ± SD, and for statistical analysis, one-way ANOVA were performed between indicated groups.

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  FIGURE 3. Cori improved glucose intolerance and insulin resistance in HFD-fed mice. (A) Levels of fasting blood glucose, and (B) levels of fasting insulin were detected in indicated groups. (C) HOMA-IR was calculated with the formula: [fasting glucose (mU/L)* fasting insulin (mmol/L)]/22.5. (D) Glucose tolerance test (GTT) was examined, and (E) the area under the curve (AUC) for GTT was calculated. (F) Insulin tolerance test (ITT) was tested, and (G) the area under the curve (AUC) for ITT was calculated.

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  FIGURE 4. Cori changed hepatic gene expression profiles in HFD-fed mice. (A) 58 down-regulated differently expressed genes (DEGs) by treatment with Cori, and (B) dendrograms of the corresponding DEGs in indicated groups. (C) 17 up-regulated differently expressed genes (DEGs), and (D) dendrograms of the corresponding DEGs by treatment with Cori in indicated groups. Group are shown in columns and genes in rows. Gene expression is indicated as a color, with brighter green and red for lower and higher values, respectively.

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  FIGURE 5. GO enrichment analysis was performed with the overlapped DEGs. (A) The categories in GO terms included cellular components (CC), molecular function (MF), and biological process (BP) were analyzed. (B) The top 20 significant GO terms of the overlapped DEGs in livers were chosen based on the order of p value from small to large.

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  FIGURE 6. KEGG pathway enrichment analysis was performed with the overlapped DEGs. (A) The enriched KEGG pathways were clustered into four main categories, including environmental information processing, human diseases, metabolism, and organismal systems, which were further divided into several subcategories. (B) The top 20 significant KEGG pathways of the overlapped DEGs in liver were selected based on the order of p value from small to large.

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  FIGURE 7. PPI network analysis was performed with the overlapped DEGs. The connections between nodes indicate the interactions between the proteins encoded by the overlapped DEGs (red nodes). The visualization for Cori-dependent hub proteins were selected with a PPI score over 0.9, such as Cyp7a1, Hsd3b5, Cyp17a1, Cyp2c38, Cyp2b9, Egfr, Erbb4, Cyp2c40, Gna14, Chrm1, Cspg5, and Gpc1.

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  FIGURE 8. Relative mRNA expression of hub genes via qPCR analysis. Compared with HFD group, The mRNA expression of Hsd3b5, Cyp2c40, Cspg5 in HCR group were increased; by contrast, the mRNA expression of Cyp7a1, Cpy17a1, Cyp2c38, Cyp2b9, Egfr, Erbb4, Gna14, Chrm1, and Gpc1 in HCR group were decreased.

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  FIGURE 9. The protective roles of Corilagin against NAFLD development in HFD-fed mice. HFD feeding induced NAFLD, which was accompanied by hepatic steatosis, high NAFLD activity score (NAS), liver injury, abnormal lipid metabolism and glucose homeostasis; however, Cori treatment significantly attenuated the progress of NAFLD.