摘要
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ECV30 4分别培养在对照组和含 5 .5mmol/L、11.1mmol/L、2 2 .0mmol/L、33.0mmol/L葡萄糖的培养基中。经葡萄糖培养 2 4h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 .0 0 0 ) ;一氧化氮浓度呈浓度依赖性增加 (r=0 .6 98,P =0 .0 0 0 ) ;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 .6 4 5 ,P =0 .0 0 9) ;小凹蛋白 1的表达也呈浓度依赖性增加 (r=0 .80 8,P =0 .0 0 0 )。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。
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.
出处
《中国动脉硬化杂志》
CAS
CSCD
2004年第1期38-42,共5页
Chinese Journal of Arteriosclerosis
