葡萄糖对血管内皮细胞小凹蛋白1和血管内皮生长因子表达的影响

为了探讨高浓度葡萄糖损伤血管内皮细胞及其对小凹蛋白-1和血管内皮生长因子表达的影响。将人脐静脉内皮细胞株ECV304．分别培养在对照组和含5.5mmol／L、11.1mmol／L、22.0mmol／L、33.0mmol／L葡萄糖的培养基中。经葡萄糖培养24h后，噻唑蓝法测定细胞增殖活性，硝酸还原酶法测定培养上清液中一氧化氮浓度，免疫组织化学和免疫印迹方法检测细胞中小凹蛋白-1和血管内皮生长因子的表达。结果发现，随着葡萄糖浓度的增加，内皮细胞增殖活性呈浓度依赖性抑制(r=-0.776，P=0.000)；一氧化氮浓度呈浓度依赖性增加(r=0.698，P=0.000)；小凹蛋白-1和血管内皮生长因子为棕黄色颗粒，主要分布于胞浆中；血管内皮生长因子的表达呈浓度依赖性增加(r=0.645，P=0.009)；小凹蛋白-1的表达也呈浓度依赖性增加(r=0.808，P=0.000)。提示高糖可诱导血管内皮细胞的血管内皮生长因子和小凹蛋白-1的表达，此变化可能与糖尿病患者高糖致血管病变有关。

Influence of Glucose on Expression of Caveolin-1 and Vascular Endothelial Growth Factors in Vascular Endothelial Cells

Aim To investigate the expression of caveolin-1 and vascular endothelial growth factors( VEGF) in vascular endothelial cells(VEC) induced by high glucose. Methods The human umbilical vein endothelial cells(ECV304) were cultured in the media containing glucose 0.0 mmol/L and 5.5, 11.1, 22.0, 33.0 mmol/L respectively. After 24 hour, the proliferation of VEC was determined by a colorimetric 3-4,5-dimethyl thiazol-2-yl]-2,5 -diphenyltetrazolium bromide (MTT) assay. The level of nitric oxide (NO) was measured by nitrate reductase method. The localization of caveolin-1 and VEGF protein was examined by immunohistochemistry. Western bolt was used to measure the protein level of caveolin-1 and VEGF. Results The proliferation of VEC was inhibited in concentration-dependent manner (r=-0.776,P=0.000). The level of NO was increased in concentration-dependent manner (r=0.698,P=0.000). Caveolin-1 and VEGF were granular brown substance which located mainly in the cytoplasm. The expression of VEGF was induced by high glucose in concentration-dependent manner(r=0.645,P=0.009). High glucose stimulated caveolin-1 expression in concentration-dependent manner(r=0.808,P=0.000). Conclusion High glucose could inhibit proliferation of vascular endothelial cells in vitro. High glucose upregulated the expression of VEGF and caveolin-1 on vascular endothelial cells, which might taken part in development of vascular complications in diabetic.