缺氧对人脐静脉内皮细胞株增殖与活力的影响  

Influence of hypoxia on the proliferation and activity of human umbilical vein endothelial cells

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

机构地区：[1]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆 400038

出　　处：《中华烧伤杂志》2007年第2期130-132,共3页Chinese Journal of Burns

基　　金：基金项目：国家重点基础研究发展规划（2005CB522601）：国家自然科学基金重点项目（10332060）

摘　　要：目的 了解缺氧对人脐静脉内皮细胞增殖与活力的影响。方法 将体外培养的人脐静脉内皮细胞株EA．hy926分为常氧组和缺氧组。常氧组常规培养，缺氧组细胞充人混合气体（含体积分数94％N2、5％CO2、1％O2）后置于37℃缺氧培养1、3、6、12h。提取两组细胞总蛋白，用蛋白质印迹法检测血管内皮生长因子（VEGF）和增殖细胞核抗原（PCNA）的表达水平；以噻唑蓝法检测细胞活力，并用流式细胞仪检测细胞周期。结果与常氧组比较，缺氧组细胞培养1hVEGF表达增高，6h达高峰，12h降低但仍明显高于常氧组；培养3h时细胞PCNA蛋白表达开始升高，6h达峰值并持续至12h。培养1、3h，缺氧组细胞活力较常氧组明显增高（P〈0．05），6h开始下降，至12h时低于常氧组但差异无统计学意义（P〉0．05）。缺氧组细胞培养1、3、6h，G0／G1期细胞均较常氧组少，S期和G2／M期细胞增多，增殖指数（P1）各为（43±9）％、（39±11）％、（40±11）％，均高于常氧组（32±9）％，其中3、6h时差异有统计学意义（P〈0．05）；12h时P1为（27±4）％，降至常氧组水平以下（P〈0．05）。结论 缺氧早期可促进人脐静脉内皮细胞增殖，但随着缺氧时间的延长细胞增殖将受抑制。Objective To investigate the influence of hypoxia on the proliferation and activity of human umbilical vein vascular endothelial cells （ EA. hy926 ）. Methods EA. hy926 cells were cultured in vitro and divided into normal control and hypoxia groups. The cells in hypoxia group were placed into hypoxic jar and treated with mixed gases（ 94% N2 ＋5%CO2 ＋1%O2） for 1,3,6 and 12 hours. Then the total proteins were extracted for the determination of the expression of vascular endothelial growth factor （VEGF） and proliferation cell nuclear antigen （PCNA）. The cell cycle and growth curve were determined with flow cytometry and MTT method, respectively. Results The expression of PC NA protein began to increase at 3 post-hypoxia hour（PHH） , peaked at 6 PHH, but without obvious difference compared with that at 12 PHH. The expression of VEGF began to increase at 1 PHH, peaked at 6 PHH, and decreased at 12 PHH, though it was still markedly higher than that of normoxia at 12 PHH. MTT results showed that the cell activity began to increase at 1 PHH, and it was still to increased at 3 PHH, then decreased at 6 PHH, and it was lower than that in control group at 12 PHH. The number of cells in G0/G1 phase was decreased, but the cells in S and G2/M phase was increased at 1, 3, 6 PHH when compared with those in normal controls. The proliferation index （P1） of cells in hypoxia group at 1PHH（43 ±9）%, 3PHH（39 ±11）%, 6 PHH（40±11）% were higher than that before hypoxia （32±9）% and3 （39 ±11）% and 6 hours （40±11）% after hypoxia （ P 〈 0.05 ）. The P1 was obviously lower at 12 PH H （ 27 ± 4 ） % compared with that of cells under normoxic condition（ P 〈 0. 05）. Conclusion Short-term hypoxia is beneficial to promote the proliferation of the cells, but this effect will be inhibited with the prolongation of hypoxia.

关 键 词：细胞低氧 内皮细胞 细胞增殖 

分 类 号：R686[医药卫生—骨科学]