高糖和缺氧对血管内皮细胞Amot表达的影响及其促血管新生功能研究

Effect of high glucose and anoxia on Amot expression in vascular endothelial cells with regard to its function in promoting angiogenesis

目的观察高糖和缺氧对血管内皮细胞（VECs）Amot表达的影响，并探讨该蛋白在血管新生中的作用。方法VECs以不同浓度葡萄糖培养48h后，再分别在正常氧或缺氧条件下培养24h，以Western印迹法测定p130-Amot和p80-Amot的蛋白表达。用siRNA下调Amot的蛋白表达，采用划痕实验及血管生成实验评价该蛋白表达下降对血管新生功能的影响。结果p130-Amot在正糖（5．5mmol／L）常氧组和正糖缺氧组明显高于高糖（30mmol／L）常氧组、高糖缺氧组、中糖（15mmol／L）常氧组和中糖缺氧组（均P〈0．01），而正糖常氧组比正糖缺氧组更高（P〈0．01）。p80-Amot在各组间的表达差异无统计学意义（P〉0．05）。用siRNA技术下调Amot蛋白表达后，VECs迁移能力和血管生成能力较正常细胞明显减弱。结论高糖比缺氧更显著降低VECsAmot蛋白的表达，该蛋白表达的下降明显抑制VECs新生血管的形成。

Objective To observe the effects of high glucose and anoxia on Amot expression in vascular endothelial ceils （ VECs ） , and explore its role in angiogenesis. Methods VECs were incubated with different glucose concentrations for 48 h, and then cultured at normal oxygen concentration or anaerobic condition for 24 h. The protein expressions of pl30-Amot and p80-Amot were detected by Western blot. After Amot expression was downregulated in VECs by siRNA, wound healing experiments and angiogenesis experiments were performed to test the effect of decreased Amot expression on angiogenesis. Results pl30-Amot protein expressions in low glucose （5.5 mmol/L） plus normal oxygen group and low glucose plus anaerobic group were higher than those in high glucose （30 mmol/L） plus normal oxygen group, high glucose plus anaerobic group, middle glucose （ 15 mmol/L） plus normal oxygen group, and middle glucose plus anaerobic group （ all P〈 0.01 ）. Compared with low glucose plus anaerobic group, p130-Amot expression was higher in low glucose plus normal oxygen group （P〈0. 01 ）. However, the expression of p80-Amot showed no statistically significant difference among different groups （ P〉0.05 ）. Compared to the normal VECs, the cells with decreased Amot expression by siRNA exhibited an attenuated cell migration in the wound healing experiments and a lesser tube formation in the angiogenesis experiments. Conclusions High glucose exerts a more significantly negtive effect on the Amot expression than anoxia in VECs. The downregulation of Amot expression inhibits migration and angiogenesis of VECs.