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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 observe the changes of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in human brain microvascular endothelial cells (HBMEC) effect. Methods: HBMEC was cultured as target cells and cultured in endothelial cell medium to prepare cell injury model. The cell viability was detected by MTT assay and the protein expressions of ICAM-1 and VCAM-1 were detected by immunocytochemical staining. Results: Compared with the normal control group, the activities of ICAM-1 and VCAM-1 were significantly increased (P <0.01) at 24, 48 and 72 h of high glucose treatment and in a dose-and time-dependent manner . CONCLUSION: High glucose has a significant damage on HBMEC. The increased expression of adhesion molecules may play a major role in the early stage of diabetic microangiopathy. The dynamic changes and toxic effects reflect the formation and the process of HBMEC.