| 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 |
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| 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） |
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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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