Objective To investigate the effect and mechanism of Fuzheng Huaxian Prescription on hepatic fibrosis mice based on PI3K/AKT/BAD signaling pathway. Methods A mouse model of hepatic fibrosis was established by intraperitoneal injection of 0.2 mL CCl4 solution (20％ in olive oil) three times per week for four consecutive weeks. C57B/L male mice were randomly divided into normal group,model group,Anluo Huaxian Pills group(0.78 g·kg-1),low-,medium-,and high-dose Fuzheng Huaxian Prescription groups(3.1,6.2,12.4 g·kg-1,respectively),with six mice in each group. The corresponding drug(10 mL·kg-1)was administered by gavage once a day for four weeks. A small-animal ultrasound device was used to measure the value of liver elasticity. Hematoxylin and eosin(HE)staining,Masson's trichrome staining and Sirius red staining were used to observe pathological changes in liver tissues. Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of hyaluronic acid(HA),laminin(LN),procollagen Ⅲ(P-ⅢC),and collagen Ⅳ(Ⅳ-C). Real-time quantitative polymerase chain reaction (qRT-PCR) and Western Blot were used to measure the mRNA and protein expression levels of protein kinase B (AKT),Bcl-2-associated agonist of cell death(BAD),the anti-apoptotic protein Bcl-2,and the pro-apoptotic protein Bax in liver tissues. Immunofluorescence was used to detect α-SMA expression in liver tissues. Results Compared with the normal group,the body mass and liver mass of rats in the model group were significantly reduced (P<0.05,P<0.01). There was significant increase in the value of liver elasticity (P<0.01),the proportion of collagen fiber area (P<0.01),serum levels of HA,LN,P-ⅢC,and Ⅳ-C(P<0.01). The proportion of α-SMA positive expression area in liver tissue significantly increased (P<0.01). The mRNA and protein expressions of AKT and Bcl-2 were significantly up-regulated (P<0.01),while those of BAD and Bax were significantly down-regulated (P<0.01). Compared with the model group,the treatment groups exhibited significant increase in the body mass and liver mass (P<0.05,P<0.01),significant reduction in the value of liver elasticity (P<0.01),and a decrease in the proportion of collagen fiber area (P<0.05,P<0.01). Serum levels of HA,LN,P-ⅢC,and Ⅳ-C were significantly reduced (P<0.01). The proportion of α-SMA positive expression area in liver tissue significantly decreased(P<0.01). Bcl-2 mRNA and protein expression were obviously down-regulated(P<0.01). AKT protein expression was significantly down-regulated (P<0.01). The mRNA expressions of BAD and Bax were obviously up-regulated (P<0.01). AKT mRNA expression in medium-and high-dose Fuzheng Huaxian Prescription groups was significantly down-regulated(P<0.01),but Bax protein expression was significantly up-regulated(P<0.01). BAD protein expression in low-and high-dose Fuzheng Huaxian Prescription groups was remarkably up-regulated (P<0.01). Conclusion Fuzheng Huaxian Prescription can significantly improve interstitial deposition of intrahepatic fibrous in hepatic fibrosis mice induced by CCl4,possibly by regulating downstream Bcl-2 and Bax protein expression through PI3K/AKT/BAD signaling pathway,promoting the apoptosis of activated hepatic stellate cells(HSC),as well as reducing extracellular matrix(ECM)and collagen fiber synthesis.
维普
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