基于法尼醇X受体通路探讨壮药依山红对胆汁淤积大鼠的影响  

Exploring the effects of Zhuang medicine Yishanhong on cholestasis in rats via the farnesoid X receptor pathway

作　　者：赵心怡 唐秀松 苏华[2] 李汶玲 揭洁 林雪婷 罗宇东[3] 庞宇舟 ZHAO Xinyi;TANG Xiusong;SU Hua;LI Wenling;JIE Jie;LIN Xueting;LUO Yudong;PANG Yuzhou(Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学壮医药学院,南宁530200 [2]广西中医药研究院 [3]广西中医药大学百年乐制药有限公司

出　　处：《环球中医药》2024年第10期1955-1962,共8页Global Traditional Chinese Medicine

基　　金：南宁市科学研究与技术开发计划项目(20213010);全国名老中医药专家传承工作室(国中医药人教函[2022]75号);广西岐黄学者培养项目(桂中医药科教发[2022]10号);广西中医药多学科交叉创新团队(GZKJ2304);广西中医药大学研究生教育创新计划项目(YCBXJ2022005)。

摘　　要：目的探讨壮药依山红对α-萘异硫氰酸酯(alpha-naphtyl isothiocyanate,ANIT)诱导胆汁淤积大鼠的影响及其作用机制。方法采用120只SD大鼠,随机分成6组,分别为正常组、模型组、熊去氧胆酸组及壮药依山红高、中、低剂量组,每组20只。除正常组外,余用ANIT灌胃建立胆汁淤积大鼠模型。连续给药15天后,称量计算大鼠肝脾指数;血清生化法检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,AKP)等转氨酶指标,测定总胆红素(total bilirubin,TBIL)、直接胆红素(direct bilirubin,DBIL)、总胆汁酸(total bile acids,TBA)等胆红素指标;苏木素—伊红(hematoxylin-Eosin,HE)染色法评估肝组织病理学变化,油红O染色评估肝脏脂滴代谢情况;蛋白免疫印迹法及实时荧光定量聚合酶链式反应检测法尼醇X受体(farnesoid X receptor,FXR)、钠牛磺胆酸共转运肽(Na+-taurocholate co-transporting polypeptide,NTCP)、多药耐药相关蛋白2(multidrug resistance-associated protein 2,MRP2)、胆汁盐转运蛋白(bile salt export pump,BSEP)蛋白和mRNA表达水平。结果与正常组比,模型组大鼠肝脾指数、血清转氨酶(ALT、AST、AKP)及胆红素(TBIL、DBIL、TBA)显著升高(P<0.05),肝组织受损。壮药依山红各剂量及熊去氧胆酸显著降低这些指标(P<0.05),改善肝组织学病变,减少脂滴沉积,效果呈剂量依赖性。同时,依山红各剂量组大鼠肝组织中NTCP、MRP2蛋白的表达量均明显升高(P<0.05);高剂量组大鼠肝组织中BSEP蛋白相对表达量显著下降(P<0.05)。与模型组相比,依山红高、中剂量组大鼠肝组织中Fxr、Ntcp、Mrp2 mRNA的相对表达量显著升高(P<0.05);依山红低剂量组大鼠仅Mrp2 mRNA表达量显著升高(P<0.05);依山红各剂量大鼠肝组织中Bsep mRNA表达水平显著下降(P<0.05)。结论壮药依山红可通过调节FXR及其调控的胆汁转运相关�Objective To investigate the therapeutic effect of Zhuang medicine Yishanhong on alpha-naphthyl isothiocyanate(ANIT)-induced cholestasis in rats,as well as to elucidate its mechanism of action.Methods A total of 120 Sprague-Dawley rats were randomly allocated into six groups:normal group,model group,ursodeoxycholic acid(UDCA)positive control group,and high-,medium-,and low-dose groups of Yishanhong.Each group consisted of 20 rats.Except for the normal group,the others were gavaged with ANIT to establish a cholestasis rat model.After 15 days of continuous administration,the liver and the spleen indices were calculated.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and alkaline phosphatase(AKP)in serum were measured.Bilirubin biomarkers,including total bilirubin(TBIL),direct bilirubin(DBIL),and total bile acids(TBA),were quantified.The pathological changes in the liver were observed through hematoxylin-eosin(HE)Staining,while the accumulation of hepatic lipid droplets was assessed using Oil Red O(ORO)Staining.The protein expression and mRNA expression levels of farnesol X receptor(FXR),Na+-taurocholate cotransporting peptide(NTCP),multidrug resistance-associated protein 2(MRP2),and bile salt transporter protein(BSEP)were assessed via Western blot and quantitative real-time polymerase chain reaction.Results Compared to the normal group,the model group rats exhibited significantly elevated liver and the spleen indices,serum transaminases(ALT,AST,AKP),and bilirubin levels(TBIL,DBIL,TBA)(P<0.05),accompanied by liver tissue damage.Zhuang medicine Yishanhong at various doses and UDCA significantly reduced these indices(P<0.05),improved histological liver lesions,and decreased lipid droplet deposition,with a dose-dependent effect.Moreover,the expression levels of NTCP and MRP2 proteins in the liver tissues of rats treated with Yishanhong at various doses were significantly increased(P<0.05);specifically,the relative expression of BSEP protein was markedly decreased in the high-dose Yishanhong

关 键 词：依山红 胆汁淤积 Α-萘异硫氰酸酯 法尼醇X受体 钠牛磺胆酸共转运肽 多药耐药相关蛋白2 胆汁盐转运蛋白 壮药 

分 类 号：R285.5[医药卫生—中药学]