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目的:研究氧化应激诱导的内皮细胞micro RNA的表达变化。方法:ECM(Endothelial Cell Medium)培养人脐静脉内皮细胞,利用不同浓度双氧水(0μmol/L,200μmol/L,500μmol/L,800μmol/L)刺激24小时后应用流式细胞术检测其凋亡水平。提取细胞总RNA,利用实时定量PCR(Quantitive real-time PCR;q RT-PCR)检测micro RNA表达量变化,并利用生物信息学软件预测可能的靶基因。结果:加入不同浓度双氧水处理24 h后的内皮细胞总凋亡率均显著高于对照组,200μmol/L、500μmol/L和800μmol/L组的凋亡率分别为(13.31%vs 4.75%,35.9%vs 4.75%,89.75%vs 4.75%,P<0.01)。200μmol/L的双氧水处理内皮细胞后,micro RNA的表达出现了明显的改变。其中mi R-92a、mi R-126的表达明显下调(P<0.05),mi R-181a、mi R-217、mi R-34a和mi R-320的表达明显上调(P<0.05)。靶基因预测显示mi R-320、mi R-92a可能调控多个和内皮细胞凋亡相关的基因表达。结论:在氧化应激诱导的内皮细胞凋亡中,mi RNA表达发生改变并可能参与调控内皮细胞功能。
Objective: To investigate the oxidative stress-induced changes of microRNA expression in endothelial cells. Methods: Human umbilical vein endothelial cells (ECMs) were cultured in ECM (Endothelial Cell Medium). The apoptosis of ECs was detected by flow cytometry after being stimulated with different concentrations of hydrogen peroxide (0μmol / L, 200μmol / L, 500μmol / L, 800μmol / L) . The total RNA was extracted and the changes of micro RNA expression were detected by qRT-PCR. Bioinformatics software was used to predict the possible target genes. Results: The total apoptosis rate of endothelial cells treated with H2O2 at different concentrations for 24 h was significantly higher than that of the control group. The apoptosis rates of 200 μmol / L, 500 μmol / L and 800 μmol / L groups were (13.31% vs 4.75%, 35.9 % vs 4.75%, 89.75% vs 4.75%, P <0.01). After treated with 200μmol / L H2O2, the expression of micro RNA changed significantly. The expressions of mi R-92a and mi R-126 were significantly downregulated (P <0.05), and the expressions of mi R-181a, mi R-217, mi R-34a and mi R-320 were significantly upregulated (P <0.05). Target gene prediction shows that mi R-320 and mi R-92a may regulate multiple gene expressions related to endothelial cell apoptosis. Conclusion: The expression of miRNA changes in oxidative stress-induced endothelial cell apoptosis and may be involved in the regulation of endothelial cell function.