基础研究(Basic Research)

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

Published at: 2017年第26卷第6期

汪海波 1 , 朱健 1 , 郗二平 1 , 张瑜 1 , 王正 1 , 谢彪 1 , 朱水波 1
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
通讯作者 水波 朱 Email: zhutian29@163.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2017.06.011
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(ROCK I/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ROCK I siRNA、ROCK II siRNA、TGF-β1单独处理、转染ROCK I siRNA或ROCK II siRNA后加TGF-β1处理的HA-VSMCs中ROCK I和ROCK II蛋白表达;比较TGF-β1单独处理、转染ROCK I siRNA或ROCK II siRNA后加TGF-β1处理以及用ROCK I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ROCK I蛋白表达明显升高(P<0.05),而ROCK II蛋白表达无明显变化(P>0.05);ROCK I siRNA或ROCK II siRNA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ROCK I蛋白表达升高被ROCK I siRNA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ROCK I siRNA转染及Y-27632处理明显抑制(均P<0.05),而不被ROCK II siRNA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ROCK I siRNA、ROCK II siRNA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:ROCK I可能在TGF-β1诱导的HA-VSMCs迁移中起主要作用,而ROCK I和ROCK II可能均不参与TGF-β1诱导的HA-VSMCs增殖。


Effects of ROCK I/II gene down-regulation on TGF-β1 induced migration and proliferation in human aortic vascular smooth muscle cells

Abstract

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 proliferation of HA-VSMCs.


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引用

引用本文: 海波 汪, 健 朱, 二平 郗, 瑜 张, 正 王, 彪 谢, 水波 朱. 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[J]. 中国普通外科杂志, 2017, 26(6): 735-741.
Cite this article as: WANG Haibo, ZHU Jian, XI Erping, ZHANG Yu, WANG Zheng, XIE Biao, ZHU Shuibo . Effects of ROCK I/II gene down-regulation on TGF-β1 induced migration and proliferation in human aortic vascular smooth muscle cells[J]. Chin J Gen Surg, 2017, 26(6): 735-741.