MiR-130a/p53通路影响动脉平滑肌细胞凋亡的实验研究

Experimental study of miR-130a/p53 pathway regulates artery smooth muscle cells apoptosis

目的探讨miR-130a引起动脉平滑肌细胞凋亡的具体机制及其潜在的临床价值。方法以组织块贴壁法原代细胞培养获得动脉平滑肌细胞,采用蛋白电泳和细胞流式分析检测到过表达miR-130a会引起细胞凋亡的变化,生物信息学GO通路富集和KEGG功能富集分析显示miR-130a跟p53通路相关,通过蛋白电泳加以验证。结果在动脉平滑肌细胞中过表达miR-130a会引起细胞抗凋亡指标bcl2表达减少,凋亡指标bax表达增加,Annexin V/PI凋亡试剂盒染色后细胞流式仪检测结果一致,蛋白电泳结果显示可以引起细胞p53的磷酸化增加。结论miR-130a可能通过促进p53的磷酸化引起动脉平滑肌细胞的凋亡增加,具有潜在的治疗价值。

Objective To investigate the specific mechanism of apoptosis in arterial smooth muscle cells induced by miR-130a and its potential clinical value.Methods Vascular smooth muscle cells were obtained by culturing with tissue block adherency method,and the overexpression of miR-130a was detected by western blotting and cell flow cytometry,which would induce cell apoptosis.Bioinformatics GO and KEGG pathway enrichment analysis showed that miR-130a was related to p53 pathway,which was verified by western blotting.Result Overexpression of miR-130a in vascular smooth muscle cells resulted in decreased expression of anti-apoptotic protein bcl2 and increased expression of apoptotic protein bax.The results of flow cytometry after Annexin V/PI apoptosis kit staining were consistent with western blotting,and the results of western blotting showed that the phosphorylation of p53 protein increased in cells.Conclusion MiR-130 a might induce the increase of apoptosis of vascular smooth muscle cells by upregulating the phosphorylation of p53,which has the potential therapeutic value.