miR-217负调控转化生长因子β受体Ⅱ抑制肝纤维化发生及发展的研究  

Study of the negative regulation of transforming growth factor beta type II receptor to inhibit the occurrence and development of liver fibrosis with miR-217

作　　者：郭义威[1] 庞盼姣 孙永琨[1] Guo Yiwei;Pang Panjiao;Sun Yongkun(Department of Human Anatomy and Histoembryology,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]新乡医学院人体解剖与组织胚胎学系,新乡453003

出　　处：《中华肝脏病杂志》2022年第7期752-757,共6页Chinese Journal of Hepatology

基　　金：新乡医学院海外博士科研启动基金(505018)。

摘　　要：目的观察miR-217对血管紧张素Ⅱ(AngⅡ)诱导的肝星状细胞(HSC)活化的影响,观察四氯化碳(CCl4)诱导小鼠中miR-217过表达产生的效果,阐明miR-217在肝纤维化中的作用。方法使用AngⅡ刺激HSC并检测miR-217表达水平的变化情况。将HSC分为对照组、AngⅡ处理组和AngⅡ+miR-217处理组,检测各组中α-平滑肌肌动蛋白,成纤维细胞特异蛋白1和Ⅰ型胶原蛋白(Collagen I)的表达水平。采用TargetScan和双荧光素酶报告基因实验对miR-217的靶基因进行筛选及验证。荧光实时定量PCR和Western blot检测miR-217对转化生长因子β受体Ⅱ(TGFBR2)表达水平的影响。构建CCl4诱导的小鼠肝纤维化模型,Masson染色和天狼星红染色检测过表达miR-217对CCl4小鼠肝纤维化进程的影响。2组数据间的比较采用t检验,多组数据比较采用One-Way ANOVA进行统计分析。结果AngⅡ刺激HSC细胞能下调miR-217的表达水平,转染miR-217 mimics能抑制AngⅡ诱导的α-平滑肌肌动蛋白,成纤维细胞特异蛋白1和Collagen I的表达水平。miR-217能特异性结合TGFBR2的3’-UTR,转染miR-217 mimic能抑制HSC中TGFBR2的mRNA和蛋白表达水平。CCl4小鼠中过表达miR-217能抑制Collagen I和CollagenⅢ的表达水平,缓解肝纤维化进程。结论miR-217通过靶向调控TGFBR2调节肝纤维化,抑制AngⅡ诱导的HSC活化,减缓CCl4小鼠的肝纤维化进程,表明miR-217可能具有抑制肝纤维化的功能。Objective To observe the effect of miR-217 on angiotensin II(AngII)-induced hepatic stellate cells(HSCs)activation,and carbon tetrachloride(CCl4)-induced overexpression in mice,so as to clarify miR-217 role in liver fibrosis.Methods HSCs were stimulated with AngⅡand the changes condition in the expression level of miR-217 were detected.HSCs were divided into control group,AngII-treated group and AngⅡ+miR-217-treated group.The expression levels of alpha-smooth muscle actin,fibroblast-specific protein 1 and collagenⅠ(CollagenⅠ)in each group were detected.The target gene of mir-217 was screened and verified by Targetscan and Dual luciferase gene reporter assay.Real-time quantitative PCR and Western blot were used to detect the effect of miR-217 on the expression level of transforming growth factor beta typeⅡreceptor(TGFBR2).A CCl4-induced mouse liver fibrosis model was constructed.Masson staining and Sirius red staining were used to detect the effect of miR-217 overexpression on the progression of liver fibrosis in CCl4 mice.Data of two groups were compared using t-test.Data of multiple groups were statistically analyzed with one-way ANOVA.Results The expression level of miR-217 was downregulated by AngⅡ-stimulated HSC cells.The expression levels ofα-smooth muscle actin,fibroblast-specific protein 1 and CollagenⅠinduced by AngⅡwas inhibited by miR-217 mimics transfection.The 3'-UTR of TGFBR2 had specifically bind miR-217.The mRNA and protein expression levels of TGFBR2 was inhibited with miR-217 mimics transfection in HSCs.The overexpression of miR-217 had inhibited the expression levels of CollagenⅠandⅢin CCl4 mice and alleviated the progression of liver fibrosis.Conclusion miR-217 regulates liver fibrosis by targeting TGFBR2,inhibits AngII-induced HSC activation,and slows down the process of liver fibrosis in CCl4 mice,suggesting that miR-217 may have an inhibitory effect on liver fibrosis.

关 键 词：肝纤维化 肝星状细胞 血管紧张素Ⅱ miR-217 转化生长因子β受体Ⅱ 

分 类 号：R575.2[医药卫生—消化系统]