miR-182在肺动脉高压患者中的表达分析及对HPVSMC细胞增殖的影响

Expression of miR-182 in patients with pulmonary hypertension and its effects on cell proliferation of HPVSMC

目的研究miR-182在肺动脉高压(PAH)患者的差异性表达,探索其在PAH中的诊断及治疗中的辅助作用。方法收集该院2017年2月至2018年2月就诊的PAH患者30例为试验组,并选取同期健康体检者30例为对照组,采用荧光素酶报告基因检测miR-182,并研究与肌细胞增强因子2C(MEF2C)靶向关系,采用Western blot方法验证miR-182对MEF2C蛋白的调控作用,采用实时荧光定量PCR方法检测miR-182及MEF2C的相对表达水平,采用CCK8方法检测miR-182对肺动脉平滑肌细胞(HPVSMC)细胞增殖的作用。结果 miR-182靶向结合于MEF2C的3′UTR区;与对照组比较,miR-182在试验组中高表达(0.963±0.148 vs.2.347±0.220,P=0.006),而MEF2C在试验组中低表达(1.013±0.179 vs.0.413±0.068,P=0.035);下调miR-182导致MEF2C蛋白表达增高(P=0.024);下调miR-182可抑制HPVSMC细胞增殖能力(P=0.019)。结论 miR-182在PAH患者中上调,并通过调节MEF2C促进平滑肌细胞增殖,miR-182及MEF2C表达改变在PAH发病机制中起到一定的作用。

Objective To research the differential expression of miR-182 in pulmonary arterial hypertension(PAH)and explore the role in the diagnosis and treatment of PAH.Methods The peripheral blood samples were collected from 30 PAH patients in Jinqiu hospital of Liaoning Province from Feb 2017 to Feb 2018.Concurrent 30 healthy people were selected as the control group.Luciferase reporter gene assay was used to detect miR-182 and investigate the targeting relationship with myocyte enhancement factor 2C(MEF2C).Western Blot was used to verify the regulatory effect of miR-182 on MEF2C protein.Real-time quantitative fluorescence PCR(RT-qPCR)was conducted to detect the relative expression of miR-182 and MEF2C.CCK8 method was used to detect the effect of miR-182 on promoting the proliferation of HPVSMC cells.Results miR-182 targeted to the 3′UTR region of MEF2C.Compared with the control group,miR-182 high-expressed in the PAH group(0.963±0.148 vs.2.347±0.220,P=0.006),and MEF2C low-expressed in the PAH group(1.013±0.179 vs.0.413±0.068,P=0.035).Downregulation of miR-182 increased the expression of MEF2C protein(P=0.024)and inhibited the cell proliferation of HPVSMC(P=0.019).Conclusion miR-182 up-regulated in patients with PAH and promotes the proliferation of smooth muscle cells by regulating MEF2C.The changes in the expression of miR-182 and MEF2C play a vital role in the pathogenesis of PAH.