论文部分内容阅读
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 study the changes of microRNA (miRNA) expression induced by high glucose in endothelial cells. Methods: Cultured human coronary artery endothelial cells were cultured in different concentrations of D-glucose solution (0 mmol / L, 5 mmol / L, 15 mmol / L and 25 mmol / L) for 24 h and then stimulated with CCK-8 Law and flow cytometry were used to detect the growth vigor and the level of apoptosis. The total RNA was collected and the miRNA expression was detected by real-time quantitative PCR (q RT-PCR). At the same time, bioinformatics prediction software such as TargetScan and PicTar was used to predict the possible target genes. Results: The cell viability was significantly decreased in high glucose (25 mmol / L) -treated cells, which was 67.5% of that in control group (P <0.01). The apoptotic rate was 4.5-fold higher than that in control group (P <0.01). The results of QRT-PCR showed that miRNA expression was significantly disturbed. MiR-451, miR-504, miR-302d, miR-18b *, miR-198, miR-328 and miR- miR-100 *, miR-137, miR-660 and miR-217 were significantly upregulated (P <0.05). Target Gene Prediction It was found that miR-217 and miR-451 may regulate the expression of multiple genes related to endothelial function. Conclusion: In high glucose-induced endothelial cell injury, miRNA expression disorders suggest that it may be involved in endothelial cell function.