阻塞性胆汁淤积中肝脏miR-200b-3p高表达对肝细胞的保护作用

Hepatic overexpression of miR-200b-3p protects hepatocytes in patients with obstructive cholestasis

目的探讨阻塞性胆汁淤积中肝脏miR-200b-3p和胆汁酸排泌转运蛋白OATP3A1的表达及意义。方法收集2012年1月至2016年6月本院肝胆外科行肝脏手术的患者30例,其中胆汁淤积20例,对照10例。检测患者肝功能;采用RT-PCR检测肝组织miR-200b-3p和OATP3A1表达;分析miR-200b-3p与肝功能和OATP3A1相关关系;用多种细胞因子处理HepG2细胞,检测miR-200b-3p表达;采用miR-200b-3p mimic和inhibitor分别转染HepG2和PLC5细胞,检测miR-200b-3p表达;采用RT-PCR及蛋白质印迹法(Western blot)检测OATP3A1表达。结果胆汁淤积组miR-200b-3p和OATP3A1显著升高(P<0.05),且两者呈正相关关系(P<0.05),miR-200b-3p与ALP和GGT呈正相关关系(P<0.05),与TBA呈负相关关系(P<0.05);FGF19诱导miR-200b-3p升高具有时间和剂量依赖性(P<0.05);miR-200b-3p mimic转染后,OATP3A1表达显著升高(P<0.05);miR-200b-3p inhibitor转染后,OATP3A1表达显著降低(P<0.05)。结论阻塞性胆汁淤积下miR-200b-3p的升高可促进OATP3A1的表达,进而促进肝脏胆汁酸外排起到保护肝脏的作用。

Objective To investigate the expression of miR-200 b-3 p and OATP3 A1 in the liver of patients with obstructive cholestasis and their protective effects on the hepatocytes.Methods Thirty patients undergoing liver surgeries in our hospital for liver,pancreatic or intestinal malignancies or for calculi in the bile duct or pancreas from January 2012 to June 2016 were enrolled,including 20 with obstructive cholestasis and 10 without cholestasis(control group).The liver functions of the patients were examined and the expression of miR-200 b-3 p and OATP3 A1 in the liver tissues were detected using RT-PCR.The correlations of miR-200 b-3 p with the liver function and OATP3 A1 expression in the patients were analyzed.In the in vitro experiment,HepG2 cells were treated with different cytokines,and the changes in the expression of miR-200 b-3 p were detected.In both HepG2 and PLC5 cells,the effects of transfection with miR-200 b-3 p mimic or inhibitor were evaluated on miR-200 b-3 p and OATP3 A1 expression.Results Compared with those in the control patients,the expression of miR-200 b-3 p and OATP3 A1 were significantly increased in the liver tissues of patients with cholestasis(P<0.05).The expression of miR-200 b-3 p was positively correlated with OATP3 A1 expression(P<0.05)and the levels of ALP and GGT(P<0.05),but was negatively correlated with TBA(P<0.05).In the cell experiment,FGF19 treatment significantly enhanced the expression of miR-200 b-3 p in HepG2 cells in a time-and dose-dependent manner(P<0.05).Transfection of HepG2 and PLC5 cells with miR-200 b-3 p mimic caused significant upregulation of OATP3 A1 expression(P<0.05),while transfection with miR-200 b-3 p inhibitor significantly downregulated OATP3 A1 expression(P<0.05).Conclusion In patients with obstructive cholestasis,the upregulation of miR-200 b-3 p enhances OATP3 A1 expression in the liver to promote hepatic bile acid efflux and thereby offer protection of the hepatocytes.