葡萄糖对血管内皮细胞的内皮素转换酶-1b表达的影响

Influence of Glucose on Expression of Endothelin-converting Enzyme-1b in Vascular Endothelial Cells

目的 探讨高浓度葡萄糖损伤血管内皮细胞及其对内皮素转换酶 - 1b表达的影响。方法 将人脐静脉内皮细胞株ECV30 4分别培养在含 0mmol L (对照组 )、5 .5mmol L (处理组 1 )、1 1 .1mmol L (处理组 2 )、2 2mmol L (处理组 3)、33mmol L (处理组 4)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及比色法检测培养上清液中丙二醛的浓度 ,逆转录聚合酶链反应法检测细胞中内皮素转换酶 - 1bmRNA。结果 发现随着葡萄糖浓度的增加 ,内皮细胞增殖呈浓度依赖性抑制 ;一氧化氮浓度呈浓度依赖性增加 ;丙二醛浓度呈浓度依赖性增加 ;内皮素转换酶 - 1bmRNA的表达呈浓度依赖性增加。结论 高浓度葡萄糖可损伤内皮细胞并诱导血管内皮细胞的内皮素转换酶 - 1bmRNA表达 ,此变化可能与糖尿病患者高血糖致血管病变有关。

Objective To investigate high glucose induces expression of endothelin-converting enzyme-1b(ECE-1b) on vascular endothelial cells(VEC). Methods The human umbilical vein endothelial cells(ECV304) were cultured in the media containing glucose 0 mmol/L (control), 5.5,11.1,22,33 mmol/L,respectively.After 24 hours, cell proliferation was evaluated 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 content of maleic dialdehyde(MDA)was examined by colorimetric assay;The expression of endothelin-converting enzyme-1b mRNA was determined by reverse transcription-polymerase chain reaction(RT-PCR).Results The rate of proliferation of VEC was inhibited in concentration-dependent manner;The level of NO was increased in concentration-dependent manner; The content of MDA was increased in concentration-dependent manner;The expression of ECE-1b mRNA was increased in concentration-dependent manner. Conclusion High glucose upregulated the expression of ECE-1b mRNA on vascular endothelial cells, which may take part in development of vascular complications in diabetic mellitus.