The mechanism of Biejiajian pill in inhibiting hepatic stellate cell activation and alleviating liver fibrosis
ZUO Yan-jie,LI Xin,CUI Jia-lin
Abstract:
Objective To investigate the mechanism of Biejiajian pill in regulating AMP dependent protein kinase (AMPK), inhibiting hepatic stellate cell activationand liver fibrosis. Methods Healthy male SPF grade C57BL/6 mice were randomly divided into control, liver fibrosis, and Biejiajian pill group (n=8). Mice of the liver fibrosis and Biejiajian pill group were induced liver fibrosis model with a high-fat and high sugar diet combined with intraperitoneal injection of carbon tetrachloride. The Biejiajian pill group was given Biejiajian pill (1.365 g/kg) by gavage once a day, starting from the 6th week. At the 12th week, serum TG, CHO, HDL, LDL, ALT, AST, and ALP were detect, and HE staining, Sirius Red staining, and α- Smooth muscle actin(α- SMA) immunohistochemical staining were used to observe the pathological changes, collagen fiber proliferation, and hepatic stellate cells (HSCs) activation respectively. In addition, western blot was used to detect the expression and activity of AMPK protein. Results The serum ALT, AST, ALP, and LDL of mice in the liver fibrosis group increased，while TG, CHO, and HDL decreased compared to the control group (P<0.05); Compared with the liver fibrosis group, there was a significant difference in the decrease of serum ALT, AST, and ALP in the Biejiajian pill group (P<0.05), while there was no statistically significant difference in TG, CHO, HDL, and LDL (P>0.05)。HE staining showed disordered arrangement of liver cells, severe steatosis, and partial ballooning in the liver fibrosis group, accompanied by inflammatory cell infiltration and patchy necrotic lesions; Sirius red staining shows obvious fibrous tissue proliferation, α- SMA expression is positive; The expression of total AMPK and p-AMPK proteins increased compared to the control group; Compared with the liver fibrosis group, the liver pathological changes, fibrous tissue proliferation, and the positive expression level of α- SMA were decreased, and both the expression of AMPK and p-AMPK proteins were decreased (P<0.05). Conclusion Biejiajian pill inhibits HSC activation and liver fibrosis, and its mechanism is related to the inhibition of AMPK protein expression and activity.
