高糖环境对人脐静脉内皮细胞分泌血栓调节蛋白的影响

摘要

目的 探讨葡萄糖对人脐静脉内皮细胞分泌可溶性血栓调节蛋白( sTM)的影响.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不同浓度的葡萄糖组(5.5、20和40 mmol/L)和对照组(不含葡萄糖),于培养0、6、24、48和72 h不同时间点收集细胞上清液,采用ELISA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,测定凝血酶原时间(PT)和活化部分凝血括酶时间(APTT)并进行比较.结果 与对照组比较,5.5 mmol/L和20 mmol/L葡萄糖组不同培养时间对内皮细胞分泌sTM的水平没有影响,差异无统计学意义(P＞0.05)；而40 mmol/L葡萄糖组内皮细胞在培养48 h内,随着培养时间的延长,内皮细胞分泌sTM的水平逐渐升高且均高于对照组,差异有统计学意义(P＜0.05)；但在培养72 h时内皮细胞分泌sTM的水平下降,与48 h不同浓度葡萄糖组及对照组比较差异均有统计学意义(P＜0.05).5.5 mmol/L和20 mmol/L葡萄糖组不同培养时间测定APTT值与对照组比较,差异无统计学意义(P＞0.05)；40 mmol/L葡萄糖组内皮细胞在培养48 h内,随培养时间的延长,APTT逐渐缩短,与其他3组比较差异有统计学意义(P＜0.01)；培养72 h与48 h比较,APTT不再缩短反而略有延长,但差异无统计学意义(P＞0.05).4组间PT值差异均无统计学意义(P＞0.05).结论 高浓度葡萄糖可导致内皮细胞损伤,使血液凝固活性增强而诱发高凝状态.%Objective To investigate the effects of glucose on the production of sTM by human umbilical vein endo-theliat cells ( HU VEC) , and to explore the possible mechanisms of diabetic vascular complications due to damage of vein endothelial cells. Methods HUVEC cultured in vilro were divided into glucose ( Glu) group (5. 5 , 20 and 40 mmol/L) , and control group ( no glucose added). Cell supernatant was collected at different time points (0, 6, 24, 48 and 72 h) and the level of sTM was detected using ELISA (enzyme linked immunosorbent assay). Meanwhile, cell supernatants were mixed with an equal volume of plasma, and the Activated Partial Thromboplastin Time (APTT) and Prothrombin Time ( PT) were measured using an automatic coagulator. Results The level of sTM induced by Glu (40 mmol/L) increased over the time, and it was higher than that in the control group after 48 h exposure ( P < 0. 05). In contrast, the level of sTM decreased after cells exposure to Clu for 72 h (P < 0. 05). The level of APTT at a high concentration of Glu (40 mmol/L) , which decreased over the time, was lower than that in the control group (P <0. 01). There was no statistical difference in PT. There was no significant difference in sTM , APTT and PT at a glucose concentration of 5.5 and 20 mmol/L, compared to the control group. Conclusion High concentrations of Glu can damage HUVEC and lead to a high coagulation state.