利胆合剂对胆汁淤积大鼠胆汁酸代谢相关蛋白的影响  

Effect of Lidan Heji on the expression of bile acid metabolism associated proteins in cholestatic rats

作　　者：李红 丘燕燕 周俪姗[1] 江治霞[1] 汤建桥[1] 鄢素琪[1] LI Hong;QIU Yan-yan;ZHOU Li-shan;JIANG Zhi-xia;TANG Jian-qiao;YAN Su-qi(Department of Integrating Traditional Chinese&Western Medicine,Wuhan Children's Hospital(Wuhan Maternal&Child Healthcare Hospital,Tongji Medical College,Huazhong University of Science&Technology,Hubei Wuhan 430016,China;Clinical College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Hubei Wuhan 430065,China)

机构地区：[1]华中科技大学同济医学院附属武汉儿童医院/武汉市妇幼保健院中西医结合科,湖北武汉430016 [2]湖北中医药大学中医临床学院,湖北武汉430065

出　　处：《中国医院药学杂志》2021年第13期1283-1287,共5页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:81704119);武汉市卫计委青年项目(编号:WZ17Q07)。

摘　　要：目的:探讨利胆合剂对胆汁淤积大鼠的影响及其作用机制。方法:采用部分胆管结扎法构建幼龄胆汁淤积SD大鼠模型。将18只部分胆管结扎幼龄SD大鼠随机分为3组:模型组、利胆合剂组(74 g·kg^(-1)·d^(-1))、熊去氧胆酸组(60 mg·kg^(-1)·d^(-1))每组6只,另取6只未经处理幼龄SD大鼠设为正常组。术后连续7 d灌胃给药后麻醉处死大鼠,取腹主动脉血及肝肠组织,采用HE染色观察肝组织病变程度;生化检测血清中总胆汁酸(total bile acid,TBA)、总胆红素(total bilirubin,TBIL)浓度及谷草转氨酶(aspartate aminotransferase,AST)、谷丙转氨酶(alanine aminotransferase,ALT)、碱性磷酸酶(alkaline phosphatase,ALP)、谷氨酰转肽酶(glutamyl transpeptidase,GGT)活性;Western blot检测肝组织中胆酸盐输出泵(bile salt export pump,BSEP)、胆固醇7α-羟化酶(cholesterol 7α-hydroxylase,CYP7A1)及肠组织中钠依赖性胆汁酸转运体(apical sodium-dependent bile acid transporter,ASBT)、成纤维细胞生长因子15(fibroblast growth factor 15,FGF15)、法尼酯X受体(farnesoid X receptor,FXR)的蛋白表达水平。结果:与正常组相比,模型组大鼠肝组织出现空泡样细胞且排列紊乱,发生胆管增生,炎症细胞浸润的现象。血清中TBA、TBIL浓度及AST、ALT、ALP、GGT活性显著增加(P<0.05)。经利胆合剂、熊去氧胆酸治疗后,对应2组上述指标均得到明显改善(P<0.05)。模型组肝肠组织中CYP7A1、ASBT蛋白表达水平较正常组显著升高(P<0.05),BSEP、FGF15、FXR蛋白表达水平则显著降低(P<0.05)。与模型组比较,利胆合剂组及熊去氧胆酸组中上述蛋白表达水平较模型组均发生明显的回转(P<0.05)。结论:利胆合剂可改善幼龄SD大鼠胆汁淤积相关检测指标,其作用机制可能与FXR依赖的胆汁酸合成、外排及重吸收相关蛋白的调控有关。OBJECTIVE To explore the effect of Lidan Heji on cholestatic rats and elucidate its mechanism.METHODS A model of young cholestatic Sprague-Dawley(SD)rat was constructed by partial bile duct ligation.Eighteen young SD rats were randomly divided into 3 groups of model,Lidan Heji(74 g·kg^(-1)·d^(-1))and ursodeoxycholic acid(60 mg·kg^(-1)·d^(-1))(n=6 each).Another 6 rats were set as normal group.After operation,rats were sacrificed with anesthesia after 7 consecutive days of intragastric administration.And abdominal aortic blood,liver tissues and ileal tissues were harvested.Hematoxylin-eosin(HE)staining was employed for observing the lesion degree of liver tissue.The serum concentrations of total bile acid(TBA)and total bilirubin(TBIL)and the activities of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP)and glutamyl transpeptidase(GGT)were measured.Western blot analysis of bile salt export pump(BSEP),cholesterol 7α-hydroxylase(CYP7A1)for liver tissues and apical sodium-dependent bile acid transporter(ASBT),fibroblast growth growth factor 15(FGF15)and farnesoid X receptor(FXR)for ileal tissues were performed for detecting the expression levels of the proteins.RESULTS As compared with normal group,the cells appeared to be vacuolated and the arrangement of cells was disordered for liver tissues in model group.Bile duct hyperplasia and inflammatory cell infiltration were present in liver tissues.As compared with normal group,serum concentrations of TBA and TBIL spiked sharply and of the activities of AST/ALT/ALP/GGT were detected in model group(P<0.05).After the treatments of Lidan Heji and ursodeoxycholic acid,the above parameters improved in each group(P<0.05).Meanwhile,in model group,the expression levels of CYP7A1 in liver and ASBT in ileal tissues were significantly higher than those in normal group while the expression levels of BSEP in liver,FGF15 and FXR in ileal tissues declined markedly(P<0.05).The expression levels of the above proteins in Lidan Heji and ursodeoxych

关 键 词：利胆合剂 FXR通路 胆汁淤积 胆汁酸代谢 

分 类 号：R963[医药卫生—微生物与生化药学]