肿瘤坏死因子相关凋亡诱导配体参与雷帕霉素诱导血管内皮细胞功能损伤的体外研究  

Tumor necrosis factor-related apoptosis-inducing ligand participates in injury of vascular endothelial cells induced by rapamycin in vitro

作　　者：叶盛[1] 张怀勤[1] 林以诺[1] 黄伟剑[1] 张艳丽[1] 邵小琳[1] 

机构地区：[1]温州医学院附属第一医院心内科,心血管生物和基因研究所,浙江温州325000

出　　处：《中国病理生理杂志》2011年第2期254-259,共6页Chinese Journal of Pathophysiology

基　　金：温州市科技局科技项目基金资助项目(No.H20080024;No.H20100009)

摘　　要：目的:探讨雷帕霉素对内皮细胞凋亡和增殖、迁移能力的影响,以及肿瘤坏死因子相关凋亡诱导配体(TRAIL)表达水平的变化。方法:用浓度为0、1、10、100μg/L的雷帕霉素孵育内皮细胞24 h,应用CCK8法检测血管内皮细胞的增殖能力,Transwell小室和划痕试验检测细胞迁移能力,DAPI染色观察凋亡细胞核形态改变,Western blotting法检测caspase-3活性以显示血管内皮细胞的凋亡,并用Western blotting检测TRAIL在凋亡的内皮细胞中的表达。结果:雷帕霉素(1-100μg/L)能诱导血管内皮细胞凋亡并抑制其迁移能力(P<0.01)。除雷帕霉素1μg/L外,10μg/L和100μg/L雷帕霉素均能抑制内皮细胞增殖能力(P<0.01),同时雷帕霉素(10-100μg/L)使TRAIL蛋白表达增加,两者作用均呈浓度依赖性(P<0.01)。结论:雷帕霉素能诱导内皮细胞发生凋亡并抑制其增殖和迁移能力。TRAIL表达上调与雷帕霉素诱导血管内皮细胞损伤有一定的相关性。AIM： To investigate the effects of rapamycin on apoptosis, proliferation, migration ability and tumor related apoptosis inducing ligand （TRAIL） in cultured human umbilical vein endothelial cells （HUVECs）. METHODS： Cultured HUVECs were treated with rapamycin at the concentrations of 0, 1, 10 and 100 μg/L for 24 h. The cell proliferation was measured by CCK -8 method. The cell migration ability was detected by Transwell chambers and wound healing test. The apoptutic index of HUVECs was quantitatively determined by measuring the activation of caspase - 3. The morphological changes of the apoptotic cells were observed by DAPI staining. The expression of TRAIL was detected by Western blotting. RESULTS： A 24 h-incubation with rapamycin（ 1 -100 μg/L） caused significant cell loss associated with the increase in apoptosis, as quantified by the determination of caspase- 3 activity（P 〈0. 01 ） in HUVECs. Obvious apoptotic morphology was observed by DAPI staining in HUVECs incubated with rapamycin. Rapamycin at the concentrations of 1 - 100 μg/L also impaired the migration ability of HUVECs（P 〈0. 01 ）. In addition, rapamycin（ 10 - 100 μg/L） inhibited the proliferation of HUVECs, whereas rapamycin at 1 μg/L had no such effect（P 〈0. 01 ）. Rapamycin（ 10 - 100 μg/L） also induced TRAIL expression in a dose - dependent manner（P 〈 0. 01 ）. CONCLUSION： Rapamycin induces apoptosis, and inhibits the proliferation and migration of HUVECs. The up - regulation of TRAIL might be related to the injury of vascular endothelial cells caused by rapamycin.

关 键 词：雷帕霉素 内皮细胞 细胞凋亡 细胞增殖 细胞迁移 肿瘤坏死因子相关凋亡诱导配体 

分 类 号：R363[医药卫生—病理学]