血管内皮细胞EA．hy926对短期低氧的反应性研究  

The responsiveness of vascular endothelial cell line EA.hy926 to short-term hypoxia

作　　者：苏踊跃[1] 梁光萍[1] 刘友生[2] 陈建[1] 杨宗城[1] 罗向东[1] 

机构地区：[1]第三军医大学西南医院烧伤研究所 创伤烧伤复合伤国家重点实验室,重庆400038 [2]第三军医大学西南医院病理学研究所,重庆400038

出　　处：《中华实验外科杂志》2007年第11期1390-1392,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(10332060);国家"973"计划资助项目(2005CB522601)

摘　　要：目的观察短期低氧应激对血管内皮细胞的影响。方法制作血管内皮细胞低氧应激模型,采用低细胞毒性的染料Almnar Blue染色法测定常氧和低氧培养条件下的细胞活力,采用流式细胞技术检测细胞周期,采用膜联蛋白V-荧光素/碘化丙锭(Annexin V-fluorescein/PI)双标记法检测细胞凋亡,应用蛋白免疫印迹方法(Westem blot),检测细胞内缺氧诱导因子和增殖细胞核抗原的表达。结果短期低氧(1～3 h)应激可以增强血管内皮细胞活力,细胞周期显示细胞4倍体峰比例增高,随着低氧时间的延长(6～12 h),细胞4倍体峰回落至常氧水平。短暂低氧(3 h),细胞凋亡比例明显增高。蛋白免疫印迹检测显示,缺氧诱导因子-1α和增殖细胞核抗原在低氧后表达增高,与低氧反应呈时间效应依赖关系。结论短期低氧启动了以增殖和活力增高为特征的血管内皮细胞适应性反应,同时也诱导了以细胞凋亡为特征的细胞损伤性反应。Objective - To investigate the vascular endothelial response to short-term hypoxia. Methods Cultured vascular endothelial cell line （EA. hy926） was exposed to hypoxia for 0,1,3,6 and 12 h. The cell viability was detected by Alamar Blue staining, cell cycle by flow cytometry, apoptosis by Annexin V-fluorescein/PI） double labeling method, hypoxia inducible factor 1 α （ HIF-1 α） and proliferating cell nuclear antigen （PCNA） expression by Western blotting respectively under normoxia or hypoxia conditions. Results The vascular endothelial viability was enhanced, and the cell cycle analysis showed that the rate of tetraploid peak was obviously increased when cells were exposed to hypoxia for 1 to 3 h, but the rate of tetraploid peak returned to normal after 6 to 12 h of hypoxia. The apoptosis rate of cells was markedly increased after short-term hypoxia （3 h）. The expression of HIF-lα and PCNA was increased in a hypoxia time-dependent manner. Conclusion Short-term hypoxia initiates adaptive response of vascular endothelial cells ,which characterized with increased cell proliferation and viability; Meanwhile it also iαnuces cellular impaired response which characterized with apoptosis.

关 键 词：低氧 内皮细胞 增殖 脱噬作用 

分 类 号：R363[医药卫生—病理学]