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目的:研究葡萄糖对人脐静脉内皮细胞蛋白C受体(EPCR)mRNA表达的影响,以及吡格列酮的干预作用。方法:体外培养人脐静脉内皮细胞(HUVECs),分别以流式细胞术和RT-PCR技术确认HUVECs膜上EPCR的表达水平和mRNA水平的表达。再分别以含不同浓度D-葡萄糖(5、10、30、50 mmol/L)的培养基以及含吡格列酮(5、10、20μmol/L)或不含吡格列酮的高糖(50 mmol/L)培养基孵育HUVECs 24 h,行剂量和时间依赖性实验,并采用实时定量PCR技术测定HUVECs细胞EPCR mRNA的表达。结果:随着培养基D-葡萄糖浓度的增加,HUVECs培养24 h后其EPCR mRNA的表达逐渐下调。在采用吡格列酮干预后,50 mmol/L高糖处理的HU-VECs EPCR mRNA表达的下调得到明显改善。结论:(1)EPCR在HUVECs上高表达,高糖可通过下调EPCR mR-NA的表达而损伤内皮细胞功能。(2)吡格列酮可阻止高糖诱导的HUVECs EPCR mRNA表达的下调,从而保护内皮细胞功能。
Objective: To investigate the effects of glucose on the expression of protein receptor C (EPCR) mRNA in human umbilical vein endothelial cells and the effect of pioglitazone. Methods: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro. Flow cytometry and RT-PCR were used to confirm the expression of EPCR and mRNA in HUVECs. The cells were cultured in medium containing different concentrations of D-glucose (5, 10, 30 and 50 mmol / L) and high glucose (50 mmol / L) containing pioglitazone HUVECs were incubated for 24 hours in dose-and time-dependent manner. The expression of EPCR mRNA in HUVECs was determined by real-time PCR. Results: With the increase of D-glucose concentration, the expression of EPCR mRNA in HUVECs gradually decreased after 24 h. After the intervention with pioglitazone, the down-regulation of EPCR mRNA expression in HU-VECs treated with 50 mmol / L of high glucose was significantly improved. Conclusion: (1) EPCR is highly expressed in HUVECs. High glucose can impair the function of endothelial cells by down-regulating the expression of EPCR mR-NA. (2) Pioglitazone could prevent the down-regulation of EPCR mRNA expression in HUVECs induced by high glucose and thus protect the function of endothelial cells.