辛伐他汀调控RhoA对血管平滑肌细胞增殖与迁移及纤溶活性的影响

目的:探讨不同浓度辛伐他汀调控小G蛋白RhoA及下游激酶ROCK信号对人主动脉血管平滑肌细胞(HAVSMCs)增殖和迁移,以及组织纤溶酶原激活物(t-PA)和纤溶酶原激活物抑制剂-1(PAI-1)表达的影响及作用机制。方法:应用不同浓度Simvastatin(0.1μmol/L、1μmol/L、10μmol/L)处理HAVSMCs,CCK-8法和"划痕实验"分别检查细胞增殖和迁移能力,RT-qPCR检测RhoA mRNA及下游ROCK mRNA表达,应用RT-qPCR和ELISA检测纤溶活性相关t-PA/PAI-1mRNA及蛋白活性变化。结果:辛伐他汀呈浓度依赖性显著抑制HUASMCs的增殖和迁移能力。辛伐他汀能够显著抑制RhoA/ROCK信号途径,诱导PAI-1 mRNA表达下调从而抑制蛋白分泌表达,同时上调t-PA mRNA表达从而增加蛋白分泌表达,以10μmol/L辛伐他汀组处理24 h效果最为显著。结论:辛伐他汀呈浓度时间依赖性抑制RhoA/ROCK信号途径,并且上调t PA以及下调PAI-1基因转录及蛋白表达,从而促进纤溶,抑制细胞增殖和迁移。

Effects of simvastatin on cell proliferation and migration ability of HAVSMCs by regulating RhoA-induced fibrinolysis

Objective:To investigate the effect and mechanism of different concentrations of simvastatin on the proliferation and migration of human vascular smooth muscle cells (HAVSMCs) by regulating RhoA/ROCK-induced expression changes of fibrinolytic activity (t-PA/PAI-1).Methods:Cultured HAVSMCs were treated with different concentrations of simvastatin (0.1 μmol/L、1 μmol /L、10 μmol /L),and then cell proliferation and cell migration were measured by CCK-8 assay and wound healing test.The expression levels of RhoA/ROCK,t-PA/PAI-1 mRNA or protein activity were detected by RT-qPCR and ELISA.Results:The proliferation and cell migration of HAVSMCs were dramatically inhibited by simvastatin in a dose-dependent manner.After HAVSMCs were treated with 10 μmol/L simvastatin for 24 h,simvastatin could significantly inhibit the RhoA/ROCK signalling pathway and induce efficient inhibition of PAI-1 mRNA and protein production levels and the promotion of t-PA mRNA and protein production levels.Conclusion:Simvastatin can significantly promote fibrinolytic activity and inhibit the proliferation and cell migration of HAVSMCs through the inhibition of the RhoA/ ROCK signalling pathway,thereby suppressing PAI-1 mRNA and protein levels and increasing t-PA mRNA and protein levels.