高糖对人脐静脉内皮细胞GSH-PX活力、NOS及ICAM-1表达的影响

Effects of high glucose on the activity of GSHPX,the expressions of NOS and ICAM1 in human umbilical vein endothelial cells

目的:探讨体外培养条件下高糖状态对血管内皮细胞的影响。方法:培养人脐静脉内皮细胞株ECV3 0 4,分为对照组和高糖组,用分光光度比色法检测谷胱甘肽过氧化物酶(GSH PX)活力,RT- PCR法检测细胞间粘附分子1(ICAM- 1)mRNA水平,免疫细胞化学法观察内皮细胞型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)的蛋白表达。结果:高糖组GSH- PX活力明显下降,eNOS、iNOS蛋白表达明显增高,ICAM- 1mRNA水平也显著上升。结论:在较短时间内糖浓度升高可引起血管内皮细胞GSH PX活力下降,ICAM -1mRNA、eNOS和iNOS蛋白表达增高,对糖尿病血管病变早期进展起一定的作用。

Objective To investigate the effects of high glucose on human umbilical vein endothelial cells (ECV304). Methods ECV304 cells were cultured in DMEM medium with or without high glucose, followed by the classification into control group and high glucose group.The activity of GSHPX in supernatant as detected by spectrophotometry assay, the mRNA expression of ICAM-1 of the cells was determined by RTPCR, meanwhile the expressions of eNOS and iNOS in the cells were observed by immunocytochemistry. Results Compared with control group,the activity of GSHPX was significantly decreased,while the protein level of eNOS and iNOS and ICAM1 mRNA expression were found to be markedly increased( P< 0.01). Conclusions Acute high glucose stimulation may down regulate the activity of GSHPX and promote the expressions of ICAM1 mRNA and eNOS、iNOS protein in vascular endothelial cells.These changes play an important role in diabetic vascular complications in the early phase.