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.
Role of miR-126/ADAM9 in myocardial fibrosis in spontaneously hypertensive rats
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.
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万方数据
维普
