护肝片调控Src激活FXR减少胆汁酸蓄积治疗胆汁淤积性肝损伤的作用机制  

Mechanism of Hugan tablet in the treatment of cholestatic liver injury by regulating Src to activate hepatocyte FXR and reduce bile acid accumulation

作　　者：张斌[1] 孙沁梅 安梓铭 石磊[1] ZHANG Bin;SUN Qin-mei;AN Zi-ming;SHI Lei(Clinical Laboratory of Shuguang Hospital affiliated to Shanghai College of Traditional Chinese Medicine(Shanghai,200434),China)

机构地区：[1]上海中医药大学附属曙光医院检验科,上海200434

出　　处：《中西医结合肝病杂志》2025年第2期201-205,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：上海中医药大学科研项目(No.18TS080)。

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。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 byα-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:①UDCA 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.②The 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 wit

关 键 词：护肝片 类固醇受体辅激活因子 法尼醇X受体 胆汁淤积性肝损伤 胆汁淤积 

分 类 号：R575.5[医药卫生—消化系统]