葡萄糖对人脐静脉内皮细胞蛋白C受体表达的影响及吡格列酮的干预作用

Effects of glucose and pioglitazone on expression of endothelial protein C receptor in human umbilical vein endothelial cells

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。

AIM： To investigate the effects of glucose and pioglitazone on the expression of endothelial protein C receptor （EPCR） in human umbilical vein endothelial cells （HUVECs）. METHODS： Cultured HUVECs were used as target cells. The expression of EPCR in HUVECs at mRNA and protein levels was confirmed by reverse transcription - polymerase chain reaction （ RT - PCR） and flow cytometry, respectively. Different concentrations of glucose were added to the culture medium to establish the injury model of HUVECs. Pioglitazone was also added to the culture medium to observe the reversible influence. The mRNA levels of EPCR were measured by real - time quantitative PCR. RESULTS： The expression of EPCR in HUVECs was down - regulated by hyperglycemia in a dose - and time - dependent manner. Pioglitazone reversed the down - regulation of the mRNA expression of EPCR by hyperglycemia in HUVECs. CONCLUSION： HUVECs express EPCR mRNA at a high level. Hyperglycemia damages the function of endothelial cells by down - regulating the expression of EPCR. Pioglitazone attenuates the injury of HUVECs induced by hyperglycemia.