郭俊 刘宇 王忠凯 李国然 沈下贤 赵仙先.氧化应激诱导内皮细胞凋亡microRNA表达改变的研究[J].现代生物医学进展英文版,2016,16(10):1821-1824. |
氧化应激诱导内皮细胞凋亡microRNA表达改变的研究 |
Study of microRNA Change in Oxidative Stress Induced Endothelial Cells Apoptosis |
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DOI: |
中文关键词: 微小RNA 内皮细胞 凋亡 氧化应激 |
英文关键词: microRNA Endothelial cells Apoptosis Oxidative stress |
基金项目:国家自然科学基金面上项目(81370266) |
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中文摘要: |
目的:研究氧化应激诱导的内皮细胞microRNA 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(0 umol/L,200 umol/L,500 umol/L,800 umol/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;qRT-PCR)检测microRNA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,200 umol/L、500 umol/L和800 umol/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)。200 umol/L的双氧水处理内皮细胞后,microRNA
的表达出现了明显的改变。其中miR-92a、miR-126 的表达明显下调(P<0.05),miR-181a、miR-217、miR-34a 和miR-320 的表达
明显上调(P<0.05)。靶基因预测显示miR-320、miR-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,miRNA 表达发生改变并可能参与调控内皮细胞功能。 |
英文摘要: |
Objective:To determine the expression change of microRNA 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 umol/L, 500 umol/L, 800 umol/L) for 24 hours. Cells apoptosis assessed by Annexin V/PI staining and flow cytometry. Total RNA
was isolated from cells, and microRNA expression levels were detected by using quantitive real-time PCR. Bioinformatics prediction
software were applied to predict potential target genes of these miRNAs.Results:The apoptosis rate of endothelial cells increased
significantly after hydrogen peroxide stimulation. Compared with control cells, the apoptosis rate of 200 umol/L, 500 umol/L and 800
umol/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 miRNA appeared obviously disorder, of which
miR-92a, miR-126 were decreased, while miR-181a, miR-217, miR-34a and miR-320 were increased notably (P<0.05). MiR-92a and
miR-320 might regulate the expression of multiple genes related to the apoptosis of endothelial cells.Conclusion:The abnormal
expression miRNAs in HUVECs after oxidative stress induced apoptosis suggested that these miRNAs maybe involved in regulation of
endothelial cells function. |
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