摘要
目的:探讨微小RNA(miR)-126/整合素和金属蛋白酶9(ADAM9)在自发性高血压大鼠(SHR)心肌纤维化中的作用.方法:实验分为正常组、SHR组、SHR+AAV9组和SHR+miR-126-AAV9组.超声心动图检查心功能,荧光显微镜观察绿色荧光蛋白(GFP)表达情况,Masson和HE染色观察心肌组织形态并统计胶原沉积评分(CVF),qRT-PCR 检测 miR-126、ADAM9 mRNA 表达,Werstern Blot 检测 α-平滑肌肌动蛋白(SMA)、ADAM9、转化生长因子(TGF)-β1蛋白表达,双萤光素酶实验验证miR-126与ADAM9的靶向关系.结果:miR-126在SHR组大鼠心肌中低表达,ADAM9高表达.与SHR+AAV9组比较,SHR+miR-126-AAV9组心肌炎症和纤维化损伤减轻,左心室缩短分数(LVFS)、左心室射血分数(LVEF)显著增高,左心室收缩末期内径(LVESD)、左心室舒张末期内径(LVEDD)、CVF、ADAM9、TGF-β、α-SAM蛋白水平显著降低.miR-126可靶向调节ADAM9表达.结论:miR-126可能通过调节成纤维细胞活化及胶原合成,参与SHR心肌纤维化发展,且可能与靶向调控ADAM9表达有关.
Abstract
Objective:To investigate the role of microRNA(miR)-126 and AD AM9,a disintegrin and a me-talloprotease 9,in myocardial fibrosis in spontaneously hypertensive rats(SHR).Methods:The ex-perimental included:normal group,SHR group,SHR+AAV9 group,and SHR+miR-126-AAV9 group.The heart function was checked by echocardiography,and then the heart tissue was retained.The expression of green fluorescent protein(GFP)was observed with a fluorescence microscope,the myocardial tissue morphology was observed with Masson and HE staining and the collagen deposition score(CVF)was counted,the expression of miR-126 and ADAM9 mRNA was detected with qRT-PCR,the protein expression of a-smooth muscle actin(SMA),ADAM9,transforming growth factor(TGF)-β1 was detected with Western Blot,and the targeting relationship between miR-126 and ADAM9 was verified with dual luciferase experiment.Results:The expression of miR-126 was low in the myocardium of rats in the SHR group,and the expression of ADAM9 was high.Compared with those in the SHR+AAV9 group,the myocardial inflammation and fibrotic injury in the SHR+miR-126-AAV9 group were alleviated,the LVFS and LVEF were significantly increased,and the protein levels of LVESD,LVEDD,CVF,ADAM9,TGF-β,and a-SAM were significantly de-creased.miR-126 could target and regulate ADAM9 expression.Conclusion:MiR-126 may partici-pate in the development of myocardial fibrosis in SHR by regulating fibroblast activation and collagen synthesis,and may be related to the targeted regulation of ADAM9 expression.
基金项目
河南省医学科技攻关计划联合共建项目和软科学项目(LHGJ20210229)
NETL
NSTL
维普
万方数据