雷帕霉素对内皮细胞迁移和血管内皮生长因子表达的影响

目的：研究不同浓度的雷帕霉素对体外培养的人血管内皮细胞（rE）4移及血管内皮生长因子（VEGF）表达的影响。方法：用含10％胎牛血清的细胞培养基（DMEM）培养正常VE细胞，用10nM，50nM，100nM和200nM的雷帕霉素孵育vE细胞24h，Westernbloting测定雷帕霉素对VE中mTOR和VEGF表达的影响，Transwell迁移模型观察不同浓度的雷帕霉素对内皮细胞迁移影响。结果：①雷帕霉素可显著抑制VE的迁移，除了在100riM之外，基本呈浓度依赖性的。100nM雷帕霉素对VE迁移的抑制作用显著减弱（P〈0．01）。②雷帕霉素对mTOR和VEGF165的表达呈浓度依赖性的抑制；而VEGF121的表达则是先升高后降低，在100nM雷帕霉素时表达最高，远远高于该浓度雷帕霉素时VEGF165的表达，可以解释100nM雷帕霉素时VE迁移抑制显著减轻的现象。结论：雷帕霉素抑制了VEGF165的表达，并且其对VE迁移抑制的效应主要由VEGF165表达减少所介导。VEGF121的表达在一定雷帕霉素浓度范围内可显著上调，从而显著改善了雷帕霉素诱导的VEGF165表达减少所致的内皮细胞迁移抑制。

The Effects of Rapamycin on Endothelial Cell Migration and Vascular Endothelial Growth Factor Expression

Objective： To investigate the effects of different-concentration rapamycin on the migration of human vascular endothelial cells （VE） cultured in vitro and the expression of vascular endothelial growth factor （VEGF） in VE cells. Methods： Normal VE cells were cultured with dulbecco＇s modified eagle＇s medium （DMEM） containing 10% fetal calf serum （FCS）. VE cells were incubated for 24h with different concentrations ofrapamycin （including 10 nM, 50 nM, 100 nM and 200 nM, 0 nM serve as control） respectively. The expression ofmTOR and VEGF in VE was detected by western bloting, and the effects of different-concentration rapamycin on endothe- lial cell migration were assessed by Transwell transwell model. Results： （1）Rapamycin significantly inhibited VE migration in a concentration-dependent style except at 100 nM, and the inhibitory effect of rapamycin on VE migration weakened significantly at 100 nM （P〈0. 01）. （2） Rapamycin inhibitied the expression of roTOR and VEGF165 in a concentration-dependent style, but VEGF121 expression in- creased at first, and then decreased. VEGF121 had the highest expression at the 100 nM rapamycin, and VEGF121 expression was more than VEGF165 expression when VE cells were incubated with 100 nM rapamycin, which may the reason of the phenomenon that the in- hibitory effect of 100 nM rapamycin on VE migration weakened significantly. Conclusions： VEGF165 expression was inhibited by ra- pamycin in a concentration-dependent manner in endothelial cell, and rapamycin inhibited VE migration mainly by VEGF 165. VEGF 121 expression significantly increased at certain concentration of rapamycin, which improved VE migration inhibition mediated by VEGF 165 expression reduction that rapamycin induced. It would provide a new idea to solve coronary restenosis because of bad re-endothelializa- tion in the clinical post-PCI.