七叶皂苷钠对胆管结扎诱导的肝纤维化作用研究

Effect of sodium aescinate on hepatic fibrosis induced by bile duct ligation

目的探究七叶皂苷钠(SA)对胆管结扎诱导的肝纤维化作用机制。方法54只雄性SD大鼠随机均分为:假手术组(Sham组)仅行开关腹,术后每日注射等量生理盐水、生理盐水组(OJ组)行胆管结扎后术后每日注射等量生理盐水、SA组(SA组)行胆管结扎后术后每日注射SA 3.6 mg/kg。通过ELISA测定各组大鼠血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBIL)的水平。HE及Masson染色观察大鼠肝脏形态结构和胶原纤维的改变。免疫荧光检测Ⅰ、Ⅲ型胶原。免疫组化及Western blot检测eIF-4E、P-4EBP1蛋白表达。结果OJ组大鼠肝脏结构紊乱,血清中的ALT、AST、TBIL水平上升,Ⅰ、Ⅲ型胶原增多,eIF-4E、P-4EBP1在肝细胞中表达减少。SA干预的大鼠血清中ALT、AST、TBIL较OJ组下降,大鼠肝脏中Ⅰ、Ⅲ型胶原表达减少且上调OJ组大鼠肝细胞中原本下调的P-4EBP1、eIF-4E。结论七叶皂苷钠可以通过促进肝细胞中的4EBP1的磷酸化及eIF-4E的激活保护肝细胞从而减少了肝纤维化。

Objective To explore the effect of sodium aescinate(SA)on hepatic fibrosis induced by bile duct ligation.Methods 54 male SD rats were randomly divided into 3 groups.Rats in the control group(Sham)were only given equivalent normal saline(NS)injection each day after laparotomy.Rats in obstructive jaundice group(OJ group)were only given equivalent NS injection each day after operation.Rats in SA group were given SA injection at the dose of 3.6 mg/kg each day after operation.There were 18 rats in each group.The expression of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL)was detected by ELISA.The changes of morphological structure and the collagen fiber of rats’liver were revealed by HE and Masson staining.The expression of collagenⅠand collagenⅢwas detected by immunofluorescence.The expression of P-4EBP1、eIF-4E was detected by immunohistochemical and Western blot analysis.Results In OJ group,the structures of liver were disordered,the expression level of ALT,AST and TBIL in SA group significantly increased,the expression level of collagenⅠand collagenⅢsignificantly increased,and the expression of eIF-4E and P-4EBP1 in hepatocytes decreased.Compared with OJ group,the expression levels of ALT,AST and TBIL in SA group significantly decreased,the expression levels of collagenⅠand collagenⅢsignificantly decreased,and the expression of eIF-4E and P-4EBP1 in hepatocytes increased.Conclusion SA can reduce liver fibrosis by promoting the phosphorylation of 4EBP1 and activating the expression of eIF-4E to protect liver cells.