吴茱萸碱对乙型肝炎大鼠肝损伤的影响及其机制

Effect and mechanism of evodiamine on liver injury in rats with hepatitis B

目的基于脂肪酸合成酶(Fas)/脂肪酸合成酶配体(FasL)信号通路探讨吴茱萸碱(EVO)对乙型肝炎大鼠肝损伤的作用机制。方法将SD大鼠随机分为对照组(未注射病毒)、模型组、吴茱萸碱低剂量组(EVO-L,5 mg/kg)、吴茱萸碱高剂量组(EVO-H,10 mg/kg)、拉米夫定组(10 mg/kg)。在大鼠体内注射乙肝病毒(HBV),建立乙型肝炎大鼠模型。EVO均使用羧甲基纤维素钠助悬。ELISA法检测血清中肝功能指标(ALT、AST、TBIL)、肝纤维化指标(HA、LN)、炎症因子(TNF-α、IL-1β)、HBsAg、HBeAg水平;HE染色观察大鼠肝组织病理学变化;微量法检测肝组织SOD、GSH、MDA水平;PCR法检测肝组织病毒载量;Western blot检测Fas、FasL、Caspase-3蛋白表达。结果与对照组比较,模型组出现肝组织结构受损,细胞坏死,炎性细胞浸润且肝组织纤维结缔增生等病理损伤,血清中ALT、AST、TBIL、HA、LN、TNF-α、IL-1β、HBsAg、HBeAg水平上升,肝组织中MDA、病毒载量、Fas、FasL、Caspase-3蛋白表达上升,SOD、GSH活性下降(均P<0.05)。与模型组比较,EVO-L组、EVO-H组和拉米夫定组肝组织病理学变化明显好转,血清中ALT、AST、TBIL、HA、LN、TNF-α、IL-1β、HBsAg、HBeAg水平下降,肝组织中MDA、病毒载量、Fas、FasL、Caspase-3蛋白表达下降,SOD、GSH活性上升(均P<0.05)。与EVO-L组比较,EVO-H组大鼠血清中ALT、AST、TBIL、HA、LN、TNF-α、IL-1β、HBsAg、HBeAg水平下降,肝组织中MDA、病毒载量、Fas、FasL、Caspase-3蛋白表达下降,SOD、GSH活性上升(均P<0.05)。与拉米夫定组比较,EVO-L组大鼠血清中ALT、AST、TBIL、HA、LN、TNF-α、IL-1β、HBsAg、HBeAg水平上升,肝组织中MDA、病毒载量、Fas、FasL、Caspase-3蛋白表达上升,SOD、GSH活性下降(均P<0.05)。结论EVO可以通过下调Fas/FasL信号通路表达,调节肝功能,抑制肝纤维化、氧化应激和炎症反应,减轻乙型肝炎大鼠的肝损伤。

Objective To investigate the effect and its mechanism of evodiamine(EVO)on liver injury in hepatitis B rats based on the fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods SD rats were randomly divided into control group(no virus injection),model group,low-dose evodiamine group(EVO-L,5 mg/kg),high-dose evodiamine group(EVO-H,10 mg/kg),Lamivudine group(10 mg/kg).All EVOs were suspended with sodium carboxymethylcellulose.A rat model of hepatitis B was established by injecting hepatitis B virus(HBV)into rats.ELISA method was used to detect the serum levels of liver function indexes(ALT,AST,TBIL),liver fibrosis indexes(HA,LN),inflammatory factors(TNF-α,IL-1β),HBsAg and HBeAg.HE staining was used to observe the pathological changes of rat liver.The levels of SOD,GSH and MDA in liver tissue were detected by micromethod.The viral load in liver tissue was detected by PCR.Western blot was used to detect the expression of Fas,FasL and Caspase-3 proteins.Results Compared with control group,the liver tissue structure was damaged in model group,with pathological damage such as cell necrosis,inflammatory cell infiltration and connective hyperplasia of liver tissue,the serum levels of ALT,AST,TBIL,HA,LN,TNF-α,IL-1β,HBsAg and HBeAg were increased,the MDA level,the viral load,and the expression of Fas,FasL,Caspase-3 proteins in liver tissue were increased,and the activities of SOD and GSH were decreased(all P<0.05).Compared with model group,the pathological changes of liver tissue were obviously improved in EVO-L group,EVO-H group and lamivudine group,the serum levels of ALT,AST,TBIL,HA,LN,TNF-αand IL-1βwere decreased,the MDA level,the viral load,and the expression of Fas,FasL,Caspase-3 proteins in liver tissue were decreased,and the activities of SOD and GSH were increased(all P<0.05).Compared with EVO-L group,the serum levels of ALT,AST,TBIL,HA,LN,TNF-αand IL-1βwere decreased in EVO-H group,the MDA level,the viral load,and the expression of Fas,FasL,Caspase-3 proteins in liver tissue were decreased,and the activities of SOD and GSH were increased(all P<0.05).Compared with Lamivudine group,the serum levels of ALT,AST,TBIL,HA,LN,TNF-αand IL-1βwere increased in EVO-L group,the MDA level,the viral load,and the expression of Fas,FasL,Caspase-3 proteins in liver tissue were increased,and the activities of SOD and GSH were decreased(all P<0.05).Conclusion EVO can regulate the liver function,inhibit the liver fibrosis,the oxidative stress and the inflammatory reaction,and alleviate the liver injury in hepatitis B rats by decreasing the expression of Fas/FasL signaling pathway.