【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。更多还原

【Abstract】 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 apoptotic 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.更多还原