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
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.