文章摘要
刘宇 曹清心 于曼丽 张必利 石屹城 崔玲 赵仙先.高糖诱导内皮细胞损伤microRNA表达紊乱的体外研究[J].,2015,15(13):2405-2407
高糖诱导内皮细胞损伤microRNA表达紊乱的体外研究
In Vitro study of MicroRNA Change in High-glucose Induced EndothelialCell Injury
  
DOI:
中文关键词: MicroRNA  内皮细胞  高糖  损伤
英文关键词: MicroRNA  Endothelial cell  High-glucose  Injury
基金项目:国家自然科学基金项目(81370266;81170223),上海市自然科学基金项目(134119a0301;13ZR1450300)
作者单位
刘宇 曹清心 于曼丽 张必利 石屹城 崔玲 赵仙先 第二军医大学附属长海医院心内科第二军医大学附属长海医院中医科 
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中文摘要:
      目的:研究高糖诱导的内皮细胞损伤微小RNA(microRNA,miRNA)的表达变化。方法:常规培养的人冠状动脉内皮细胞,利 用不同浓度D- 葡萄糖溶液(0 mmol/L、5 mmol/L、15 mmol/L和25 mmol/L),诱导刺激24 h 后分别用CCK-8法和流式细胞术检测 其生长活力和凋亡水平。收集细胞总RNA,利用实时定量PCR(Quantitative real-time PCR,qRT-PCR)检测miRNA的表达变化,同 时利用TargetScan、PicTar 等生物信息学预测软件预测可能的靶基因。结果:高糖溶液(25 mmol/L)刺激内皮细胞后,细胞生长活 力明显降低,为对照组的67.5 %(P<0.01),凋亡水平为对照组的4.5 倍(P<0.01)。QRT-PCR 结果显示miRNA 的表达出现了明显 的紊乱,其中miR-451、miR-504、miR-302d、miR-18b*、miR-198、miR-328 和miR-517c 明显下调,miR-29c、miR-100*、miR-137、 miR-660 和miR-217 明显上调(P<0.05)。靶基因预测发现miR-217 和miR-451 可能调控内皮细胞功能相关的多个基因的表达。结 论:在高糖诱导的内皮细胞损伤中,miRNA 表达紊乱提示其可能参与内皮细胞功能。
英文摘要:
      Objective:To determine the expression change of microRNA (miRNA) in endothelial cell injury induced by high-glucose.Methods:Human coronary artery endothelial cells cultured by conventional methods, were stimulated by using different concentrations of D-glucose solution (0 mmol/L, 5 mmol/L, 15 mmol/L and 25 mmol/L). Cells vitality and apoptosis were measured by CCK-8 assay and flow cytometry. Total RNA was isolated from cells in 24h after high-glucose induced, and miRNA expression levels were detected by using quantitative real-time PCR. Bioinformatics prediction software including Targetscan and PicTar etc, were applied to predict potential target genes of these miRNAs.Results:Endothelial cell viability decreased significantly after high-glucose (25 mmol/L) stimulation, with 67.5%to the control cells (P<0.01), and the apoptosis level were 4.5 times compared with control cells (P<0.01). Quantitative real-time PCR results showed that the expression of miRNA appeared obvious disorder, which were miR-451, miR-504, miR-302d, miR-18b*, miR-198, miR-328 and miR-517c were increased, while miR-29c, miR-100*, miR-137, miR-660 and miR-217 were decreased notably (P<0.05). MiR-217 and miR-451 might regulate the expression of multiple genes related to endothelial cell function by Bioinformatics prediction.Conclusion:The abnormal expression miRNAs in endothelial cells after high glucose induced injury suggested that these miRNAs maybe be involved in regulation of endothelial cells function.
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