【摘要】 目的:探究护肝片通过调控酪氨酸蛋白激酶类固醇受体辅激活因子(Src)激活肝细胞法尼醇X受体(FXR),从而减少胆汁酸蓄积治疗胆汁淤积性肝损伤(CLI)的作用机制。方法:将雄性健康SD大鼠42只随机分为正常组，模型组，熊去氧胆酸(UDCA)组，护肝片低、中、高剂量(10、20、40 mg/kg)组，护肝片+PP2组，每组6只；各组分别给予对应药物或溶剂灌胃，1次/d,连续7 d;第8天除正常组外其余6组均给予75 mg/kg α-萘异硫氰酸酯(ANIT)诱导建立大鼠胆汁淤积模型，护肝片+PP2组建模后腹腔注射1.5 mg/kg p-Src抑制剂PP2溶液，1次/d,连续2 d。采用酶标仪检测大鼠血清肝功能指标；采用苏木精-伊红(HE)染色观察肝组织病理学变化；采用Western Blotting检测护肝片对CLI大鼠肝组织中Src、p-Src、FXR蛋白表达的影响；检测PP2对CLI大鼠血清肝功能指标含量的影响；HE染色检测p-Src/Src蛋白对CLI大鼠肝组织形态学的影响；免疫共沉淀实验检测FXR与Src蛋白间的相互作用；Western Blotting检测PP2对p-Src/Src及FXR蛋白表达的影响。结果:(1)UDCA组和护肝片低、中、高剂量组均不同程度地降低ANIT引起的大鼠血清肝功能指标升高；与模型组比较，UDCA组及护肝片低、中、高剂量组大鼠肝组织形态学有明显改善；与正常组比较，模型组大鼠肝组织中p-Src/Src、FXR蛋白表达明显降低；与模型组比较，UDCA组及护肝片中、高剂量组大鼠肝组织中p-Src/Src、FXR蛋白表达明显上升。(2)与模型组比较，护肝片高剂量组大鼠血清肝功能指标明显下降；与护肝片高剂量组比较，护肝片+PP2组大鼠血清肝功能指标明显增加；与模型组比较，p-Src抑制剂PP2可明显抑制护肝片对大鼠肝组织形态学的改善。结论:护肝片可以通过调控Src磷酸化激活大鼠肝细胞FXR受体，使胆汁酸蓄积减少，从而治疗CLI。更多还原

【Abstract】 Objective:To explore the mechanism of Hugan tablet regulating steroid receptor coactivator(Src)to activate farnesoic X receptl(FXR) in hepatocytes to reduce bile acid accumulation in the treatment of cholestatic liver injury.Methods:The animal model of cholestatic liver injury in SD rats induced by α-naphthalene isothiocyanate(ANIT) was used and randomly divided into normal group, model group, ursodeoxycholic acid(UDCA) group, and low-, medium-and high-dose(10 mg/kg, 20 mg/kg, 40 mg/kg) groups of Hugan tablet.The serum biochemical indexes of rats were detected by enzyme-linked immunosorbent assay; the pathological changes of liver tissue were observed by HE staining; the effects of Hugan tablet on the expression of Src, p-Src and FXR proteins in liver tissue of CLI rats were detected by Western Blotting; the effects of p-Src inhibitor PP2 on the serum index content of CLI rats were detected; the effects of p-Src/Src protein on the morphology of liver tissue of CLI rats were detected by HE staining; the interaction between FXR and Src proteins were detected by Co-IP experiment; the effects of p-Src inhibitor PP2 on the expression of p-Src/Src and FXR proteins were detected by Western Blotting.Results:(1)UDCA group and Hugan tablet 10, 20, 40 mg/kg groups all reduced the increase of serum biochemical indexes caused by ANIT to varying degrees; the liver tissue morphology of rats in UDCA group and Hugan tablet 10 mg/kg, 20 mg/kg, 40 mg/kg groups were significantly improved compared with the model group; compared with the normal group, the expression of p-Src/Src and FXR proteins in liver tissue of rats in model group were significantly decreased, and the expression of p-Src/Src and FXR proteins in liver tissue of rats in UDCA group and Hugan tablet 20 mg/kg, 40 mg/kg groups were significantly increased compared with the model group.(2)The serum index content of rats in Hugan tablet group was significantly decreased compared with the model group, and the serum index content of rats in Huganpian +PP2 group was significantly increased compared with the Hugan tablet group. Compared with the model group, the p-Src inhibitor PP2 could significantly inhibit the improvement of liver tissue morph.Conclusion:The FXR receptor in rat hepatocytes is activated through the regulation of Src phosphorylation by Hugan tablet, leading to a reduction in bile acid accumulation and potentially providing therapeutic benefits for cholestasis-induced liver injury(CLI).更多还原