间充质干细胞移植干预2-OA/BSA诱导的原发性胆汁性胆管炎小鼠肝内胆管上皮细胞自噬蛋白表达的变化  

Mesenchymal stem cell transplantation alleviating hepatic injury by modulating intrahepatic biliary epithelial cell autophagic flux in mice with 2-O A-BSA–induced primary biliary cholangitis

作　　者：朱赟[1] 姚根宏[1] 唐小军[1] Zhu Yun;Yao Genhong;Tang Xiaojun(Department of Rheumatology and Immunology,Affiliated Drum Tower Hospital,Nanjing University Medical School,Nanjing 210008,Jiangsu Province,China)

机构地区：[1]南京大学医学院附属鼓楼医院风湿免疫科,南京市210008

出　　处：《实用肝脏病杂志》2019年第3期337-340,共4页Journal of Practical Hepatology

基　　金：国家自然科学基金资助项目(编号:81401348);"十三五"南京市卫生青年人才培养工程计划项目(编号:QRX17040)

摘　　要：目的观察异基因间充质干细胞(MSCs)移植干预原发性胆汁性胆管炎(PBC)小鼠对肝内胆管上皮细胞(IBECs)自噬的调控作用及其分子机制。方法将30只C57BL/6小鼠随机分为模型组(n=18)、BSA组(n=6)和未处理组(n=6)。采用2-辛炔酸结合牛血清白蛋白(2-OA-BSA)注射诱导PBC模型。再将PBC模型小鼠随机分为模型组(PBC组,n=4)、MSCs移植干预组(MSCs组,n=6)和转录激活因子3(STAT3)抑制剂干预组(Stattic组,n=6)。采用灌流法分离肝内胆管树纯化IBECs,采用WB法检测IBECs组织STAT3/pSTAT3、p62、LC3、PKR/pPKR、Beclin-1蛋白、e IF2α/peIF2α、LAMP-1表达,采用RT-PCR法检测STAT3、LC3和p62-m RNA,使用电镜观察IBECs内自噬泡形成。结果模型组肝组织汇管区淋巴细胞浸润并有散在的肉芽肿形成,而MSCs和Stattic干预组小鼠汇管区炎性细胞浸润减少;模型组小鼠IBECs自噬蛋白Beclin-1、STAT3和pSTAT3表达较BSA组增强,而MSCs移植和Stattic干预组上述蛋白表达减弱;模型组和MSCs组p62 m RNA水平较BSA组下降,模型组STAT3 m RNA水平较BSA组下降。结论我们成功建立了2-OA-BSA诱导的PBC小鼠模型,MSCs移植干预通过下调STAT3表达减轻了PBC小鼠肝组织汇管区损害,可能与调控小鼠体内自噬相关蛋白表达有关。Objective To establish a reliable animal model of primary biliary cholangitis(PBC) and to investigate the therapeutic effect of umbilical cord-derived mesenchymal stem cells (UC-MSC) on STAT3 signal in intrahepatic biliary epithelial cells(IBECs) of this PBC animal. Methods C57BL/6 mice were intraperitoneally injected with 2-octynyl acid (2-OA)-bovine serum albumin (BSA) adjuvanted with Freund's adjuvant/incomplete Freund's adjuvant (CFA/IFA) or with same amount of BSA adjuvanted with IFA,or untreated as control. The model mice 22 weeks later were randomly divided into model (n=4),MSCs transplantation (n=6) and STAT3 inhibitors-treated group(n=6). The intrahepatic biliary tree and IBECs were obtained,and STAT3/pSTAT3,p62,LC3,PKR/pPKR,Beclin-1,eIF2α/peIF2α and LAMP-1 expression were detected by WB,and STAT3,LC3 and p62-mRNA were detected by RT-PCR. Results The lymphocyte infiltration and granuloma in portal area were found in liver tissues of model mice,while they were obviously alleviated in MSCs-or Stattic-intervened groups;the expressions of Beclin-1,STAT3 and pSTAT3 in IBECs from model intensified as compared to those in BSA-treated group,while they were weaken in MSCs-or Stattic-intervened groups;p62 mRNA levels decreased in model and MSC transplanted grous as compared to that in BSA-treated group,and STAT3 mRNA level in model decreased as compared to that in BSA-treated group. Conclusion Our findings indicate that MSC transplantation might regulate the autophagy and decrease the expression of STAT3 signals in mice with primary biliary cholangitis.

关 键 词：原发性胆汁性胆管炎 间充质干细胞 自噬 STAT3信号通路 

分 类 号：R575.7[医药卫生—消化系统]