摘要
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 determine the expression change of micro RNA in endothelial cells under oxidative stress. Methods：Human Umbilical Vein Endothelial Cells cultured by ECM, were stimulated by different concentrations of hydrogen peroxide（0 umol/L,200 μmol/L, 500 μmol/L, 800 μmol/L） for 24 hours. Cells apoptosis assessed by Annexin V/PI staining and flow cytometry. Total RNA was isolated from cells, and micro RNA expression levels were detected by using quantitive real-time PCR. Bioinformatics prediction software were applied to predict potential target genes of these mi RNAs. Results： The apoptosis rate of endothelial cells increased significantly after hydrogen peroxide stimulation. Compared with control cells, the apoptosis rate of 200 μmol/L, 500 μmol/L and 800μmol/L hydrogen peroxide treatment group were significantly increased（13.31 % vs 4.75 %, 35.9 % vs 4.75 %, 89.75 % vs 4.75 %,respectively, P〈0.01）. Quantitive real-time PCR results showed that the expression of mi RNA appeared obviously disorder, of which mi R-92 a, mi R-126 were decreased, while mi R-181 a, mi R-217, mi R-34 a and mi R-320 were increased notably（P〈0.05）. Mi R-92 a and mi R-320 might regulate the expression of multiple genes related to the apoptosis of endothelial cells. Conclusion： The abnormal expression mi RNAs in HUVECs after oxidative stress induced apoptosis suggested that these mi RNAs maybe involved in regulation of endothelial cells function.
出处
《现代生物医学进展》
CAS
2016年第10期1821-1824,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金面上项目(81370266)
关键词
微小RNA
内皮细胞
凋亡
氧化应激
microRNA
Endothelial cells
Apoptosis
Oxidative stress
