基于FXR通路研究獐牙菜调控ANIT诱导的肝损伤大鼠胆汁酸代谢效应机制

Swertia bimaculata Regulates Bile Acid Metabolism in ANIT-Induced Liver Injury in Rats via FXR Signaling Pathway

目的:研究獐牙菜对α-异硫氰酸萘酯(ANIT)诱导的胆汁淤积型肝损伤大鼠的保护作用。方法:将40只雄性SD大鼠分为正常对照组、模型对照组、熊去氧胆酸100 mg/kg组、獐牙菜194、385 mg/kg组。采用ANIT诱导大鼠急性肝内胆汁淤积型肝损伤模型,以獐牙菜进行干预,熊去氧胆酸为阳性对照,观察大鼠体质量及肝质量变化;测定胆汁流速,检测其血清ALT、AST、ALP、γ-GT活力,DBIL、TBIL、TBA含量;HE染色观察肝组织病理变化;qRT-PCR检测肝组织Fxr、Mrp2、Bsep、Ntcp、Cyp7a1和Shp mRNA的表达。结果:与正常对照组相比,模型对照组大鼠在ANIT干预后状态不佳,体质量逐日下降,胆汁流速显著下降(P<0.01);血清中AST、ALT、ALP、γ-GT、TBA、TBIL、DBIL活力或含量显著增加,肝脏系数显著增加(P<0.01);肝组织病理损伤明显,肝组织Fxr、Mrp2、Bsep、Ntcp和Shp的mRNA表达均明显下调,Cyp7a1的mRNA表达明显上调(P<0.05或P<0.01);与模型对照组相比,獐牙菜各剂量组及熊去氧胆酸100 mg/kg组中肝脏系数显著下降,肝组织病理损伤显著减轻,胆汁流速显著加快,血清中ALT、AST、ALP、γ-GT活力,TBIL、TBA和DBIL含量不同程度降低,Fxr和Bsep的mRNA表达上调,Cyp7a1的mRNA表达下调(P<0.05或P<0.01);獐牙菜385 mg/kg组和熊去氧胆酸100 mg/kg组Mrp2和Ntcp的mRNA表达上调,獐牙菜各组大鼠肝脏中的Shp mRNA表达上调(P<0.05或P<0.01)。结论:獐牙菜具有保护ANIT诱导的胆汁淤积型肝损伤大鼠肝损伤的作用。

Objective:To study the protective effect of Swertia bimaculata(Sieb.et Zucc.)Hook.f.et Thoms.ex C.B.Clark(SBH)on cholestatic liver injury induced byα-naphthyl isothiocyanate(ANIT)in rats.Methods:Forty male SD rats were randomly assigned into a normal control group,a model group,an ursodeoxycholic acid(UDCA,100 mg/kg,positive control)group,and SBH(385 and 194 mg/kg)groups.The rat model of acute intrahepatic cholestasis was established with ANIT.The changes in body weight and liver weight were measured.The flow velocity of bile and the serum levels of alanine transaminase(ALT),aspartate transaminase(AST),alkaline phosphatase(ALP),gamma-glutamyltransferase(-GT),direct bilirubin(DBIL),total bilirubin(TBIL),and total bile acid(TBA)were determined.Hematoxylineosin(HE)staining was employed to observe the histopathological changes of liver in each group.The mRNA levels of Fxr,Mrp2,Bsep,Ntcp,Cyp7al,and Shp were determined by qRT-PCR.Results:Compared with the normal control group,the model group showed poor conditions,decreased body weight and bile flow rate,elevated serum levels of AST,ALT,ALP,γ-GT,TBA,TBIL,and DBIL,and increased liver index.In addition,the model group showed obvious pathological damage in the liver tssue,with down-regulated mRNA levels of Fxr,Mrp2,Bsep,Ntcp,and Shp and up-regulated mRNA level of Cyp7al in the liver.Compared with the model group,SBH and UDCA decreased the liver index,alleviated the pathological damage in the liver,accelerated the bile flow,and lowered the serum levels of ALT,AST,ALP,γ-GT,TBIL,TBA,and DBIL.Moreover,they up-regulated the mRNA levels of Fxr and Bsep and down-regulated the mRNA level of Cyp7al.In addition,SBH(385 mg/kg)and UDCA up-regulated the mRNA levels of Mrp2 and Ntcp and all doses of SBH up-regulated the mRNA level of Shp.Conclusion:SBH can protect rats from the cholestatic hepatic injury induced by ANIT.