MicroRNA-484通过靶向肝星状细胞中Fis1调控肝纤维化进程  

MicroRNA-484 regulates liver fibrosis course through targeting Fis1 in hepatic stellate cells

作　　者：张利芬[1] 李彬彬[1] 余宏宇[1] 

机构地区：[1]第二军医大学长征医院病理科,上海200003

出　　处：《第二军医大学学报》2017年第9期1146-1151,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81370553)~~

摘　　要：目的探讨微RNA-484(miR-484)对肝纤维化发生和发展过程中关键细胞肝星状细胞(HSC)的作用和影响。方法以前期芯片筛选结果为基础,通过细胞转染人为干预miR-484在HSC-T6细胞株中的表达。利用qPCR、蛋白质印迹法检测肝纤维化及凋亡相关指标的表达变化;以AnnexinⅤ-FITC/PI双染细胞后,用流式细胞仪检测miR-484对细胞凋亡的影响。结果与对照组相比,人为下调miR-484在HSC-T6细胞株中的表达后,其靶基因Fis1及促凋亡因子caspase-3在mRNA和蛋白水平的表达均升高(P<0.05),凋亡抑制因子Bcl-2表达下降(P<0.05);流式细胞仪检测到HSC-T6细胞凋亡率升高[(32.81±3.21)%vs(57.54±6.76)%,P<0.05];同时肝纤维化相关指标α-平滑肌肌动蛋白、Ⅰ型胶原的mRNA和蛋白水平均下调(P<0.05)。结论 miR-484通过靶向Fis1抑制HSC凋亡、促进其活化,从而促进肝纤维化的发生和发展。Objective To explore the effects of microRNA-484（miR-484）on hepatic stellate cells（HSCs）,the key cell in the occurrence and development of liver fibrosis,and to investigate the functions.Methods On the basis of our previous microarray analysis results,we transfected HSC-T6 cell lines with miR-484 inhibitor to intervene the expression of miR-484 in vitro.The expressions of mRNA and proteins related to liver fibrosis and apoptosis were detected by qPCR and Western blotting,respectively.The cell apoptosis with AnnexinⅤ-FITC/PI double staining was determined by flow cytometry.Results Compared with the control group,the mRNA and protein expression of miR-484-targeted gene Fis1 and proapoptotic factor caspase-3were both significantly up-regulated（P〈0.05）,and apoptosis inhibitory factor Bcl-2 was significantly down-regulated（P 〈0.05）in the HSC-T6 cells transfected with miR-484inhibitor;the apoptosis rate of HSC-T6 cells was significantly increased（[32.81±3.21]% vs[57.54±6.76]%,P〈0.05）,andα-smooth muscle actin and collagen typeⅠ were both significantly down-regulated（P〈0.05）in the HSCT6 cells transfected with miR-484 inhibitor.Conclusion MiR-484 promotes the occurrence and progress of liver fibrosis through inhibiting the apoptosis and promoting the activation of HSCs by targeting Fis1.

关 键 词：微RNA-484 线粒体分裂蛋白1 肝纤维化 肝星状细胞 活化 细胞凋亡 

分 类 号：R575.2[医药卫生—消化系统]