葡萄糖和胰岛素对人脐静脉内皮细胞蛋白C受体分泌水平的影响

Effects of glucose and insulin on production of solubility endothelial cell protein C receptor in human umbilical vein endothelial cells

目的:探讨在葡萄糖和胰岛素作用下人脐静脉内皮细胞可溶性内皮细胞蛋白C受体(sEPCR)的分泌,阐明内皮功能损伤与糖尿病血栓形成的机制。方法:将体外培养的人脐静脉内皮细胞分为葡萄糖组(5.5、20.0和40.0 mmol·L-1)、胰岛素组(0.1、1.0和10.0 nmol·L-1)、高糖(40 mmol·L-1)环境下的胰岛素组(0.1、1.0和10.0 nmol·L-1)和对照组(不含葡萄糖和胰岛素),于0、6、24、48和72 h收集细胞上清液,采用ELISA双抗体夹心法检测sEPCR水平。结果:葡萄糖组内皮细胞分泌sEPCR的水平均高于对照组(P<0.01),且随着培养时间的延长逐渐升高;但高糖环境下(40 mmol·L-1)培养48 h后sEPCR的水平不再升高,反而下降(P<0.01);胰岛素组内皮细胞分泌sEPCR的水平均高于对照组(P<0.01),且随着培养时间的延长逐渐升高,但高胰岛素环境下(10 nmol·L-1)培养72 h后sEPCR的水平不再升高,反而下降(P<0.01);高糖(40 mmol·L-1)环境下的胰岛素组,不同浓度胰岛素在培养24 h内,sEPCR的水平与单独应用葡萄糖组比较均降低(P<0.01)。结论:胰岛素在一定程度内可以抵消高浓度葡萄糖对内皮细胞的损伤作用,但长期高浓度的胰岛素可以加重内皮细胞的损伤,诱发血栓形成。

Objective To investigate the effects of glucose and insulin on the production of solubility endothelial cell protein C receptor（sEPCR） in human umbilical vein endothelial cells（HUVEC） and to clarify the mechanisms of the damage of vein endothelial cells and the diabetic thrombosis.Methods HUVEC cultivated in vitro were divided into glucose（Glu） groups（5.5,20.0 and 40.0 mmol·L-1）,insulin groups（0.1,1.0 and 10.0 nmol·L-1）,insulin groups（0.1,1.0 and 10.0 nmol·L-1） in high glucose environment（40 mmol·L-1）and control group（without glucose and insulin）.The cell supernatant was collected at different time（0,6,24,48 and 72 h） and the levels of sEPCR were detected by ELISA assay.Results The levels of sEPCR of HUVEC in Glu groups were higher than that in control group （P0.01）,and it was gradually increased with the prolongation of time.But the level of sEPCR at high concentration of Glu（40 mmol·L-1） was decreased after the cells were cultured for 48 h（P0.01）.The level of sEPCR of HUVEC in insulin group was higher than that in control group（P0.01）,and and it was gradually increased with the prolongation of time.But the level of sEPCR at high concentration of Glu（40 mmol·L-1） was decreased after the cells were cultured for 72 h（P0.01）.The levels of sEPCR at high concentration of Glu with different concentrations of insulin were lower than the levels of those cultured with Glu（P0.01）.Conclusion Insulin can inhibit the damage effect of high concentration Glu on HUVEC,but long-time use of high concentration insulin can aggravate the damage of HUVEC and induce thrombosis.