ROCK Ⅰ/Ⅱ基因下调对TGF-β1诱导的人主动脉平滑肌细胞迁移及增殖的影响  

Effects of ROCK Ⅰ/Ⅱ gene down-regulation on TGF-1 induced migration and proliferation in human aortic vascular smooth muscle cells

作　　者：汪海波[1] 朱健[1] 郗二平[1] 张瑜[1] 王正[1] 谢彪[1] 朱水波[1] 

机构地区：[1]南方医科大学附属武汉临床医学院/中国人民解放军广州军区武汉总医院心胸外科,湖北武汉430070

出　　处：《中国普通外科杂志》2017年第6期735-741,共7页China Journal of General Surgery

基　　金：湖北省武汉市科技局应用基础研究计划基金资助项目(2015060101010053)

摘　　要：目的:探讨Rho激酶Ⅰ/Ⅱ(ROCKⅠ/Ⅱ)对转化生长因子β1(TGF-β1)诱导的人主动脉平滑肌细胞(HA-VSMCs)迁移及增殖的影响。方法:比较转染ROCKⅠsiRNA、ROCKⅡsiRNA、TGF-β1单独处理、转染ROCKⅠsiRNA或ROCKⅡsiRNA后加TGF-β1处理的HA-VSMCs中ROCKⅠ和ROCKⅡ蛋白表达;比较TGF-β1单独处理、转染ROCKⅠsiRNA或ROCKⅡsiRNA后加TGF-β1处理以及用ROCKⅠ抑制剂Y-27632后加TGF-β1处理的HA-VSMCs的迁移与增殖能力。实验均以无处理的HA-VSMCs为空白对照。结果:与空白对照HA-VSMCs比较,TGF-β1单独处理后HA-VSMCs的ROCKⅠ蛋白表达明显升高(P<0.05),而ROCKⅡ蛋白表达无明显变化(P>0.05);ROCKⅠsiRNA或ROCKⅡsiRNA转染后HA-VSMCs中各自靶蛋白表达明显降低(均P<0.05),TGF-β1诱导的ROCKⅠ蛋白表达升高被ROCKⅠsiRNA转染明显抑制(P<0.05)。与空白对照HA-VSMCs比较,TGF-β1单独处理后HA-VSMCs细胞迁移数明显增加(P<0.05),该作用被ROCKⅠsiRNA转染及Y-27632处理明显抑制(均P<0.05),而不被ROCKⅡsiRNA转染影响(P<0.05);TGF-β1单独处理后HA-VSMCs增殖明显增强(P<0.05),而无论ROCKⅠsiRNA、ROCKⅡsiRNA转染或Y-27632处理均对TGF-β1的增殖诱导作用无明显影响(均P>0.05)。结论:ROCKⅠ可能在TGF-β1诱导的HA-VSMCs迁移中起主要作用,而ROCKⅠ和ROCKⅡ可能均不参与TGF-β1诱导的HA-VSMCs增殖。Objective： To investigate the effects of Rho-associated coiled-coil containing protein kinase I/II （ROCK I/II） on migration and proliferation in human aortic vascular smooth muscle cells （HA-VSMCs） induced by transforming growth factor β1 （TGF-β1）. Methods： The protein expressions of ROCK I and ROCK II among HA-VSMCs respectively treated with ROCK I siRNA transfection, ROCK II siRNA transfection, TGF-β1 alone, and ROCK I siRNA or ROCK II siRNA transfection plus TGF-β1 were compared. The migration and proliferation abilities among HA-VSMCs respectively treated with TGF-β1 alone, ROCK I siRNA or ROCK II siRNA transfection plus TGF-β1, and ROCK I inhibitor Y-27632 plus TGF-β1 were compared. Untreated HA-VSMCs were used as blank control for all experiments. Results： Compared with blank control HA-VSMCs, the ROCK I protein expression was significantly increased （P〈0.05） but ROCK II protein expression was unchanged （P〉0.05） in HA-VSMCs after TGF-β1 alone treatment, the corresponding target protein expression was significantly decreased in HA-VSMCs after ROCK I siRNA or ROCK II siRNA transfection （both P〈0.05）, and the increased ROCK I protein expression in HA-VSMCs induced by TGF-β1 was significantly inhibited by ROCK I siRNA transfection （P〈0.05）. Compared with blank control HA-VSMCs, the number of migrating cells in HA-VSMCs after TGF-β1 alone treatment was significantly increased （P〈0.05）, and this effect was significantly inhibited by ROCK I siRNA transfection or Y-27632 pretreatment （both P〈0.05）, but was not influenced by ROCK II siRNA transfection （P〉0.05）; the proliferation in HA-VSMCs was significantly enhanced by TGF-β1 alone treatment, and the TGF-β1-induced proliferation was not affected by either ROCK I siRNA, ROCK II siRNA transfection or Y-27632 pretreatment （all P〉0.05）. Conclusion： ROCK I may play a major role in TGF-β1-induced migration of HA-VSMCs, but either ROCK I or ROCK II may not participate in TGF-β1-induced proli

关 键 词：主动脉 肌 平滑 血管 RHO相关激酶类 细胞运动 

分 类 号：R654.3[医药卫生—外科学]