MiR-223通过靶向PARP1对氧化应激状态下人主动脉内皮细胞HAECs存活的影响

Overexpression of miR-223 reduces survival of human aortic endothelial cells with H_2O_2-induced oxidative stress in vitro by directly targeting PARP1

目的探讨miR-223通过靶向PARP1对氧化应激状态下人主动脉内皮细胞HAECs存活的影响及miR-223改善氧化应激来治疗高血压的潜能。方法 Target Scan预测靶向PARP1的潜在miRNAs;双荧光素酶报告实验检测miR-223对PARP1的靶向调节作用;q PCR和western blot检测miR-223对HAECs细胞中PARP1 mRNA和蛋白表达影响;用H2O2诱导人主动脉内皮细胞HAECs产生氧化应激,单细胞凝胶电泳、克隆形成实验和MTT分别检测miR-223对细胞DNA损伤、存活能力的影响。结果软件预测发现miR-223是PARP1的潜在miRNAs;双荧光素酶报告实验、q PCR及western blot均证实了miR-223对PARP1的靶向调控作用;过表达miR-223可通过降低PARP1表达促进H2O2诱导人主动脉内皮细胞HAECs氧化损伤,降低细胞存活能力。结论 miR-223通过靶向调节PARP1表达降低H2O2诱导后人主动脉内皮细胞HAECs存活,有可能成为高血压治疗的新靶点。

Objective To investigate the effect of miR-223 overexpression on the survival of human aortic endothelial cells （HAECs） under oxidative stress and explore the underlying mechanism. Methods Targetscan was used to predict the potential miRNAs targeting poly（ADP-ribose） polymerase 1 （PARP1）. Dual luciferase reporter assay, qPCR and Western blotting were used to evaluate the effect of miR-223 overexpression on PARP1 in cultured HAECs transfected with miR-223 using Lipofectamine 2000. The effect of miR-223 overexpression on DNA damage and cell viability in HAECs exposed to H2O2-induced oxidative stress was investigated using single cell gel electrophoresis, clonogenic assay and MTT assay. Results Bioinformatics analysis indicated that PARP1 was a potential target of miR-223. The results of dual luciferase reporter assay, qPCR and Western blotting all supported the role of miR-223 for targeted modulation of PARP1 in HAECs. Overexpression of miR-223 obviously exacerbated oxidative cell damage and decreased the cell viability in HAECs exposed to H2O2. Conclusion The overexpression of miR-223 reduces the survival of cultured HAECs with H2O2-induced oxidative stress by targeting PARP1, suggesting the potential of miR-223 as a therapeutic target for improving hypertension induced by oxidative stress.