摘要
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 explore the expression and function of miRNA-21 and programmed cell death 4 （PDCD4） in Human umbilical vein endothelial cells （HUVECs） treated with high glucose or insulin. Methods： HUVECs were treated with D-Glucose in different concentrations （5.5mM, 11 mM, 22 mM and 33 mM） or insulin （0nM, lnM, 10nM, 50nM, 100nM, 1000nM） respectively for 2 days, and then cells were collected to detect miRNA-21 and PDCD4 mRNA expression levels with the real-time fluorescence quota RT-PCR method. The expression of protein PDCD4 in HUVECs were determined by Western blot analysis. Results： To compare with the control group of 5.5 mM glucose, miRNA-21 expressions in 11 mM, 22 mM and 33 mM groups were down-regulated significantly along with the increasing of glucose concentrations （p 〈0.01）. PDCD4 protein expression were increased（p 〈0.05）, but no difference ofPDCD4 mRNA expression was observed among four groups （p〉0.05）.To compare with the control group of 0nM insulin, miRNA-21 expression were down-regulated in high insulin groups （10nM, 50nM, 100nM and 1000nM） and miRNA-21 expression was reduced most significantly in 1000 nM insulin group （p 〈0.05）. PDCD4 protein expressions were up-regulated in high insulin groups （10nM, 50nM, 100nM and 1000nM） （p 〈0.01）, but no difference of PDCD4 mRNA expression was observed among groups （p 〈0.05）. Conclusions： The expression of miRNA-21 and PDCD4 protein were influenced in vascular endothelial cells treated by high glucose or high insulin and the signal transduction pathways involving in miRNA-21 and PDCD4 possibly participated in the process of apoptosis in vascular endothelial cells treated by high glucose or high insulin.
出处
《现代生物医学进展》
CAS
2010年第6期1076-1079,共4页
Progress in Modern Biomedicine
基金
深圳市重点资助项目(200643)资助
