益气活血利水汤调控Nrf2/GPX4信号通路抑制铁死亡改善肝纤维化大鼠的机制研究OA

Objective:To investigate the mechanism by which Yiqi Huoxue Lishui Decoction ameliorates hepatic fibrosis(HF)in rats by inhibiting hepatocyte ferroptosis via activation of the nuclear factor-E2-related factor 2(Nrf2)/glutathione peroxidase 4(GPX4)signaling pathway.Method:Forty Sprague-Dawley(SD)rats were randomly divided into five groups(n=8 per group):the control group(Control),model group(Model),traditional Chinese medicine group(TCM),traditional Chinese medicine+ferroptosis inhibitor group(TCM+Fer-1),and traditional Chinese medicine+ferroptosis inhibitor+autophagy inhibitor group(TCM+Fer-1+3-MA).The model was established by intraperitoneal injection of a 50%carbon tetrachloride(CCl4)solution(1.5 mL/kg),administered twice weekly for 4 weeks.Treatment groups received their respective interventions:the Yiqi Huoxue Lishui Decoction was administered via oral gavage(2 mL/100 g),while the ferroptosis inhibitor Ferrostatin-1(Fer-1)(1 mg/kg)and the autophagy inhibitor 3-Methyladenine(3-MA)(15 mg/kg)were administered via intraperitoneal injection.All administrations were performed once daily for 4 weeks,while the other groups were given an equivalent volume of normal saline.Hepatic histopathological changes were evaluated using hematoxylin-eosin(HE)and Masson staining.Serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD),malondialdehyde(MDA),and Fe2+content in serum and liver tissue were measured.Western blot(WB)was employed to assess protein expression of GPX4,Nrf2,solute carrier family 7 member 11(SLC7A11),transferring receptor 1(TFR1),Beclin-1,and the LC3 II/I ratio.Quantitative real-time polymerase chain reaction(qPCR)was utilized to determine mRNA expression levels of Nrf2 and GPX4.Results:Compared with the Control group,the Model group exhibited severe hepatic steatosis,significant fibrosis,elevated collagen deposition,increased serum AST,ALT,MDA,TNF-α,and Fe2+levels,and reduced SOD activity(P<0.05).Protein expression of GPX4,Nrf2,and SLC7A11 was upregulated,while TFR1 and Beclin-1 expression increased,and mRNA levels of GPX4 and Nrf2 were markedly suppressed(P<0.05).Intervention with Yiqi Huoxue Lishui Decoction and/or inhibitors significantly attenuated histopathological damage,reduced AST,ALT,MDA,TNF-α,Fe2+levels,and downregulated TFR1 and Beclin-1 expression(P<0.05).Concurrently,SOD activity,GPX4 and Nrf2 protein/mRNA expression,and SLC7A11 protein levels were elevated(P<0.05).No significant difference was observed in the LC3 II/I ratio across groups(P>0.05).Conclusion:Yiqi Huoxue Lishui Decoction may ameliorate hepatic fibrosis by inhibiting ferroptosis through activation of the Nrf2/GPX4 signaling pathway.