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目的:研究miRNA-21和程序性细胞死亡因子4(PDCD4)在不同浓度葡萄糖或胰岛素培养血管内皮细胞中的表达情况,探讨miRNA-21对内皮细胞凋亡功能的影响。方法:用不同浓度D-葡萄糖(5.5、11、22及33mM)和胰岛素(0、1、10、50、100及1000nM)分别干预人脐静脉内皮细胞(HUVECs),2天后收集细胞用实时荧光定量RT-PCR方法检测细胞中miRNA-21及PDCD4的mRNA表达情况,用蛋白印迹方法检测PDCD4在细胞中的表达变化。结果:与5.5mM葡萄糖对照组相比,高葡萄糖浓度组(11、22及33mM)中HUVECs的miRNA-21随着葡萄糖浓度的增加表达明显降低(p<0.01);PDCD4的蛋白表达增加(p<0.05),但PDCD4的mRNA表达无差异(p>0.05)。与0nM胰岛素组相比,10、50、100、1000nM胰岛素组细胞中miRNA-21表达下降(p<0.05),PDCD4的蛋白表达增加(p<0.01),1000nM组miRNA-21表达降低程度和PDCD4的蛋白表达增加程度最为明显(p<0.05),但不同浓度胰岛素对PDCD4的mRNA表达无差异(p>0.05)。1nM胰岛素对HUVECs中miRNA-21表达和PDCD4的蛋白表达无差异(p>0.05)。结论:高浓度葡萄糖或胰岛素可影响HUVECs中miRNA-21和PDCD4蛋白的表达,miRNA-21与PDCD4相关信号途径调控可能是高糖和高胰岛素影响内皮细胞凋亡的重要机制之一。
Objective: To investigate the expression of miRNA-21 and PDCD4 in different concentrations of glucose or insulin-cultured vascular endothelial cells and to explore the effect of miRNA-21 on the apoptosis of endothelial cells. Methods: Human umbilical vein endothelial cells (HUVECs) were separately treated with different concentrations of D-glucose (5.5, 11, 22 and 33mM) and insulin (0, 1, 10, 50, 100 and 1000nM). After 2 days, Quantitative RT-PCR was used to detect the mRNA expression of miRNA-21 and PDCD4 in the cells, and the expression of PDCD4 in the cells was detected by Western blotting. Results: Compared with 5.5mM glucose control group, miRNA-21 of HUVECs in high glucose group (11, 22 and 33mM) was significantly decreased with the increase of glucose concentration (p <0.01), while the expression of PDCD4 protein increased <0.05), but PDCD4 mRNA expression was no difference (p> 0.05). Compared with 0nM insulin group, miRNA-21 expression decreased (p <0.05) and PDCD4 protein increased (p <0.01) in 10,50,100,1000nM insulin group, while decreased expression of miRNA-21 in 1000nM group and PDCD4 (P <0.05). However, there was no difference in PDCD4 mRNA expression between different concentrations of insulin (p> 0.05). 1nM insulin had no difference in the expression of miRNA-21 and PDCD4 in HUVECs (p> 0.05). Conclusion: High concentration of glucose or insulin can affect the expression of miRNA-21 and PDCD4 in HUVECs. The regulation of miRNA-21 and PDCD4 signaling pathways may be one of the important mechanisms of high glucose and high insulin on endothelial cell apoptosis.