胰岛素对高糖诱导人脐静脉内皮细胞分泌血栓调节蛋白的影响  

Effects of insulin induced by hyperglycemia on the production of solubility thrombomodulin in human umbilical vein endothelial cells

作　　者：佟广辉[1] 佟威威[1] 金冬岩[1] 刘勇[1] 

机构地区：[1]中国医科大学附属盛京医院检验科,辽宁沈阳110004

出　　处：《武警医学院学报》2012年第6期428-430,438,共4页Acta Academiae Medicinae CPAPF

基　　金：辽宁省教育厅高校科学研究基金资助课题(L2010671)

摘　　要：【目的】研究胰岛素对高糖诱导的人脐静脉内皮细胞可溶性血栓调节蛋白(solubility Thrombomodulin,sTM)分泌的影响和可能机制。【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和40 mmol/L)、胰岛素组(10 IU/mL)、高糖(40mmol/L)环境下的胰岛素组(10 IU/mL)和对照组(不含葡萄糖和胰岛素),于0、6、24、48和72 h等不同时间点收集细胞上清液,采用ELISA法双抗体夹心法检测sTM水平,同时将各时间的细胞上清液与正常血浆等比例混合后测定活化部分凝血活酶时间(Activated partial thromboplastin time,APTT)。【结果】在培养的48 h内,高糖环境下(葡萄糖浓度40 mmol/L)内皮细胞分泌sTM的水平随着时间的延长逐渐升高且均高于对照组(P<0.05),但培养72 h后sTM的水平不再升高,反而下降(P<0.05),高糖环境下(葡萄糖浓度40 mmol/L),APTT值逐渐降低且均低于对照组(P<0.01);胰岛素组内皮细胞在不同培养时间,分泌sTM的水平逐渐升高且均高于对照组(P<0.05),与单独应用高糖(葡萄糖浓度40 mmol/L)相比较均降低(P<0.05),APTT值随着培养时间的延长逐渐升高,但仍低于对照组(P<0.01)。【结论】生理浓度胰岛素可降低高糖环境下内皮细胞分泌sTM的水平,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。[Objective] To determine the effects of insulin induced by hyperglycemia on the production of sTM in human umbilical vein endothelial ceils （HUVEC） and the possible mechanisms. [ Methods ] HUVEC which were cultured in vitro were divided into glucose （Glu） group （5.5,20 and 40 mmol/L）, insulin group （10 IU/mL insulin ＋ 40 mmol/L glucose ）and control group （ not including glucose and insulin）. The cell supernatant was collected at different time（0, 6,24,48 and 72 h） and the level of sTM were detected by ELISA （ enzyme linked immunosorbent assay ）. And at the same time, the effect of Activated partial thromboplastin time （APTT） mixed proportion with the cell supernatant from normal blood was assayed. [ Results] The level of sTM of HUVEC at high concentration of Glu （40 mmol/L）was higher than that of control group and it increased with time going by after the cells exposed for 48 h （P 〈 0.05）. But the level of sTM decreased after the cells exposed for 72 h（P 〈 0.05）. The level of APTT at high concentration of Glu（40 mmol/L）was lower than that of control group and it decreased with time going by（P〈 0.01）The level of sTM at high concentration of Glu with insulin was lower than the level of those cultured with GIu（P〈 0.05）. The level of APTT at high concentration of Glu with insulin increased with time going by, it was still lower than that of control group （P〈 0.01 ）. [Conclusion] Insulin can reduce sTM level and inhibit the damage effect of high concentration Gin on HUVEC.

关 键 词：人脐静脉内皮细胞 血栓调节蛋白 葡萄糖 胰岛素 

分 类 号：R967[医药卫生—药理学]