目的:探讨缺氧条件下miR-145对血管平滑肌细胞(VSMC)增殖、迁移及凋亡的影响及可能作用机制。方法:采用组织贴壁方法培养获得兔原代VSMC。用脂质体转染法将miR-145mimics转入VSMC细胞。实验设置为常氧条件下(21%O_2,5%CO_2,74%N2)空白对照组、miR-NC组、miR-145mimic组、miR-145inhibitor组;缺氧条件下(3%O_2,5%CO_2,92%N_2)空白对照组,miR-NC组,miR-145mimic组,miR-145inhibitor组。应用real time PCR检测miR-145在各组细胞表达水平。采用EdU染色检测VSMC增殖能力,Transwell实验检测VSMC细胞迁移能力,流式细胞仪检测细胞凋亡。采用targetScan,miRanda和PicTar靶基因预测软件和DAVID数据库对miR-145进行生物信息学分析,预测可能的靶基因。结果:在常氧与缺氧条件下,与miR-NC组相比,转染miR-145mimic组增殖率和迁移率明显减少,细胞凋亡率增高(P<0.05);转染miR-145inhibitor组增殖率和迁移率明显增加,细胞凋亡率下降(P<0.05)。同时与常氧条件相比,缺氧组增殖率和迁移率明显增加以及细胞凋亡率下降(P<0.05)。生物信息学分析提示miR-145靶基因的通路信号富集主要为促分裂素原活化蛋白激酶1(MAPK1),细胞周期蛋白依赖性激酶6(CDK6),钙/钙调素依赖蛋白激酶2D(Ca2+/CAMK2D)信号通路等。结论:缺氧条件下,在VSMC细胞中miR-145低表达、miR-145过表达可以有效的抑制VSMC细胞增殖、迁移及促进该细胞凋亡
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