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目的:探讨ROCKI/II在转化生长因子β1(TGF-β1)诱导的人主动脉平滑肌细胞(HA-VSMC)表型转化中的作用.方法:将HA-VSMC分别转染ROCKI和ROCKII的siRNA后荧光显微镜观察转染情况,并用Western blot方法检测不同处理的HA-VSMC(ROCKI siRNA转染、ROCKII siRNA转染、+TGF-β1、ROCKI siRNA转染+TGF-β1、ROCKII siRNA转染+TGF-β1)中ROCKI和ROCKII蛋白的表达;分别用Western blot和RT-PCR方法检测不同处理的HA-VSMC(+TGF-β1、ROCKI siRNA转染+TGF-β1、ROCKII siRNA转染+TGF-β1、ROCK非特异性抑制剂Y-27632预处理+TGF-β1)中细胞收缩表型标志物α-平滑肌肌动蛋白(α-SMA)、平滑肌22α(SM22α)与合成表型标志物骨桥蛋白(OPN)的蛋白与mRNA表达,均以无处理的HA-VSMC为空白对照.结果:免疫荧光观察与Western blot检测表明两种siRNA均成功转染;TGF-β1处理后,HA-VSMC中ROCKI蛋白表达明显升高(P0.05),ROCKI siRNA转染后TGF-β1上调ROCKI的作用被明显抑制(P<0.05).与空白对照组HA-VSMC比较,TGF-β1处理后的HA-VSMC中α-SMA、SM22α的蛋白和mRNA表达明显降低,而OPN蛋白与mRNA表达明显升高(均P<0.05),ROCKI siRNA转染或Y-27632预处理后,TGF-β1的上述作用均明显减弱(均P0.05).结论:TGF-β1可诱导HA-VSMC由收缩表型向合成型表型转化,ROCKI表达的升高可能在这一转化中起主要作用.“,”Objective: To investigate the actions of ROCKI/II in phenotypic transformation of human aortic vascular smooth muscle cells (HA-VSMCs) induced by transforming growth factor β1 (TGF-β1). Methods: HA-VSMCs were respectively transfected with ROCKI and ROCKII, and the transfection results were observed by fluorescence microscope. The ROCKI and ROCKII protein expressions in HA-VSMCs with different treatments (ROCKI siRNA transfection, ROCKII siRNA transfection, +TGF-β1, ROCKI siRNAtransfection+TGF-β1, and ROCKII siRNAtransfection+TGF-β1) were determined by Western blot analysis. The protein and mRNA expressions of the contractile phenotype maker α-smooth muscle actin (α-SMA) and smooth muscle 22α (SM22α) and synthetic phenotype marker osteopontin (OPN) in HA-VSMCs with different treatments (+TGF-β1, ROCKI siRNA transfection+TGF-β1, ROCKII siRNA transfection+TGF-β1, and pretreatment of ROCK non-specificity Y-27632+TGF-β1) were determined by Western blot analysis and RT-PCR method, respectively. Untreated HA-VSMCs were used as blank control. Results: Both siRNAs were successfully transfected as evidenced by fluorescence observation and Western blot analysis. In HA-VSMCs after TGF-β1 treatment, the ROCKI protein expression level was significantly up-regulated (P0.05), while the ROCKI increasing effect of TGF-β1 was significantly inhibited by ROCKI siRNA transfection (P<0.05). In HA-VSMCs after TGF-β1 treatment, the protein and mRNA expressions of α-SMA and SM22α were decreased and those of OPN were increased significantly (allP<0.05), and these effects were signi昀cantly suppressed by ROCKI siRNA transfection or Y-27632 pretreatment (allP0.05). Conclusion: TGF-β1 can induce the transformation of HA-VSMCs from contractile phenotype to synthetic phenotype, which may be associated with the up-regulation of ROCKI expression.