Beneficial Activities of Alisma orientale Extract in a Western Diet-Induced Murine Non-Alcoholic Steatohepatitis and Related Fibrosis Model via Regulation of the Hepatic Adiponectin and Farnesoid X Receptor Pathways

Figures

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  Figure 1. UHPLC chromatogram of the AO extract and UV spectrums of the seven peaks.

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  Figure 2. Administration of Alisma orientale (AO) extract decreases WD-induced body and liver weight gain and prevents hepatomegaly (a–f). (a) The food intake of mice was measured during the experiment period. (b) The body weight changes in mice following Western diet-induced nonalcoholic steatohepatitis (NASH). (c) The body weight gain was calculated. After the mice were sacrificed, (d) epididymis fat weights and (e) liver weights were measured. (f) Representative images of liver tissues. The data were analyzed by one-way analysis of variance (ANOVA) and (B) two-way ANOVA. *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01 vs. WD-treated group (n = 12 per group). Plus sign (+) and minus sign (−) indicate the presence and absence of oral administration of WD or AO extract in male C57BL/6 mice, respectively.

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  Figure 3. Administration of AO extract improves serum liver function, lipid profile, and glucose level (a–f). The levels of (a) alanine transaminase (ALT), (b) aspartate transaminase (AST), (c) cholesterol, (d) triglycerides (TG), and (e) low-density lipoprotein in serum. (f) Oral glucose tolerance test was assessed. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WD-treated group (n = 12 per group). Plus sign (+) and minus sign (−) indicate the presence and absence of oral administration of WD or AO extract in male C57BL/6 mice, respectively.

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  Figure 4. AO extract intervention resolves histological injury of NASH with fibrosis (a–i). (a) Representative images of H&E, picro sirius red (PSR) (n = 6 per groups), and F4/80 staining (n = 4 per groups) of mice liver tissues (scale bar = 50 μm; arrow: steatosis; bold arrow: lobular inflammation; dotted line arrow: ballooning). (b) NAFLD activity scores and (c) fibrosis scores were evaluated. The mRNA expression of (d) TIMP metallopeptidase inhibitor 1 (TIMP1), (e) platelet-derived growth factor (PDGF), (f) transforming growth factor beta 1 (TGFβ1), (g) F4/80, and the protein production of (h) tumor necrosis factor-α (TNF-α) as well as (i) interleukin-1β (IL-1β) in the liver (n = 6 per group) were evaluated. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WD-induced group. Plus sign (+) and minus sign (−) indicate the presence and absence of oral administration of WD or AO extract in male C57BL/6 mice, respectively.

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  Figure 5. Administration of AO extract affects adiponectin and farnesoid X receptor (FXR) activation. (a) Representative immunoblotting images and quantitative protein expressions for (b) adiponectin and (c) AMP-activated protein kinase (AMPK) in mice liver. (d) Representative immunoblotting images and quantitative protein expressions for (e) FXR, (f) osteopontin (OPN), (g) focal adhesion kinase (FAK), and (h) protein kinase B (AKT). The mRNA expressions of (i) phosphoenolpyruvate carboxykinase-cytosolic (PEPCK-c), (j) PEPCK-mitochondrial (PEPCK-m), (k) OPN, and (l) CYP7A1 in the mice liver. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WD-treated group (n = 6 per group). Plus sign (+) and minus sign (−) indicate the presence and absence of oral administration of WD or AO extract in male C57BL/6 mice, respectively.