高糖对人脑微血管内皮细胞黏附分子表达的影响及在内毒形成中的作用

Effect of high glucose on expression of human brain microvascular endothelial cellular adhesion molecule and formation of endogenetic toxins

目的:观察高糖环境下人脑微血管内皮细胞(HBMEC)细胞间黏附分子-1(ICAM-1)及血管细胞黏附分子-1(VCAM-1)表达的变化,探讨其在内毒形成中的作用。方法:以培养HBMEC作为靶细胞,在内皮细胞培养基中加入无水葡萄糖制备细胞损伤模型。采用四甲基偶氮唑盐法(MTT法)检测高糖环境下细胞的活性;采用免疫细胞化学染色方法检测黏附分子ICAM-1、VCAM-1的蛋白表达。结果:高糖作用24、48、72h时,与正常对照组比较活性显著降低,ICAM-1、VCAM-1的蛋白表达水平也均明显升高(P<0.01),并呈剂量、时间依赖性。结论:高糖对HBMEC有显著的损伤作用,黏附分子表达升高在糖尿病微血管病变早期可能起主要作用,其动态变化和毒性效应体现了内毒形成及作用过程。

Objective： To explore the effect of high glucose on formation of endogenetic toxins and expression of intercellular cell adhesion molecule- 1 （ICAM- 1） and vascular cell adhesion molecule- 1 （VCAM- 1） of human brain microvascular endothelial cells. Methods： Cultured HBMEC was used as target cells, anhydrous glucose was added to the cell medium to prepare the injury model of human endothelial cell. Methyl thiazolyl tetrazolium method was used to examine the cell activity in a high glucose environment. Immunocytochemistry method was used to detect the expression of ICAM-1 and VCAM-1 of HBMEC. Results： The expressions of ICAM- 1 and VCAM- lof HBMEC were enhanced by high glucose after pretreating HBMEC for 24,48 and 72h（P〈0.01, compared with the normal group）. Moreover, the damage effect enhanced as the concentration of glucose increased. Conclusions： High concentration of glucose has a significant effect of HBMEC injury. The data suggested that adhesion molecule 1 may play important role in the early state of diabetic microvascular complications. Formation and actions of endogenetic toxins are reflected by the dynamic changes and toxic effects.