高糖诱导内皮细胞损伤microRNA表达紊乱的体外研究  

In Vitro study of Micro RNA Change in High-glucose Induced Endothelial Cell Injury

作　　者：刘宇[1] 曹清心[2] 于曼丽[1] 张必利[1] 石屹城[1] 崔玲[1] 赵仙先[1] 

机构地区：[1]第二军医大学附属长海医院心内科,上海200433 [2]第二军医大学附属长海医院中医科,上海200433

出　　处：《现代生物医学进展》2015年第13期2405-2407,共3页Progress in Modern Biomedicine

基　　金：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)

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,q RT-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。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-ghicose 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）. Qtmntitative 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 ceils function.

关 键 词：MICRORNA 内皮细胞 高糖 损伤 

分 类 号：R-33[医药卫生] R587.1