VEGF165对过氧化氢诱导的内皮祖细胞增殖损伤的保护作用  

The protective effect of VEGF165 on the injury of endothelial progenitor cells proliferation activity induced by hydrogen peroxide

作　　者：刘芳[1,2] 桑甜甜[2] 曹青[2] 王飞[2] 黄黎亚[2] 郑兴[1] 

机构地区：[1]上海第二军医大学附属长海医院心内科,上海市200433 [2]上海交通大学医学院附属新华医院老年科,上海市200092

出　　处：《老年医学与保健》2014年第6期380-383,共4页Geriatrics & Health Care

基　　金：国家自然科学基金项目（81270205）

摘　　要：目的 观察血管内皮生长因子-165（vascular endothelial growth factor-165,VEGF165）对过氧化氢（hydrogen peroxide,H2O2）诱导的人外周血来源内皮祖细胞（endothelial progenitor cells,EPCs）增殖损伤及细胞周期相关蛋白表达的影响.方法 采用密度梯度离心法和贴壁培养法从人外周血中分离培养内皮祖细胞.在经MTT实验筛选出过氧化氢处理浓度后,对EPCs进行实验分组：正常对照组,不给予任何处理因素;VEGF165处理组,以终浓度为25ng/mL的VEGF165孵育24 h;过氧化氢处理,以终浓度200 μmol/L过氧化氢处理内皮祖细胞24 h;VEGF 165预处理组,经浓度为25 ng/mL的VEGF165预孵育30 min后,再与终浓度为200μmol/L的H2O2共孵育24 h.CCK-8法检测各组内皮祖细胞增殖活性的差异;Western blot分析各组内皮祖细胞中细胞周期相关蛋白cyclin D1和PCNA的蛋白表达情况.结果 与对照组相比,VEGF165处理组内皮祖细胞增殖活性增高（P＜0.05）;过氧化氢处理组内皮祖细胞增殖活性较对照组降低（P＜0.05）,且细胞周期相关蛋白cyclin Dl和PCNA下调（P＜0.05）; VEGF165预处理组中,EPCs增殖活性较过氧化氢处理组增高（P＜0.05）,且cyclin D1和PCNA蛋白表达上调（P＜0.05）.结论 VEGF165可促进人外周血来源的内皮祖细胞的增殖,对过氧化氢诱导的内皮祖细胞增殖损伤有一定的保护作用,其机制可能与细胞周期相关蛋白cyclin D1和PCNA表达升高有关.Objective To investigate the effects of VEGF165 on proliferation activity and the level of cell cycle regulatory proteins in endothelial progenitor cellsinduced by hydrogen peroxide （H2O2）. Methods Endothelial progenitor cells （EPCs） were isolated and cultured from human peripheral blood by density gradient centrifugation method and adherent culture method. Three groups, including the control group, VEGF165 treatment group, H2O2 treatment group and VEGF165 pretreatment group, were set up after the best working concentration of H2O2 was sorted out. Cell proliferation activity was assessed by CCK-8 assay. The expression changes of cell cycle regulatory proteinscyclin D1 and PCNA in endothelial progenitor cells among groupswere detected by western blot. Results Compared with the control group, the proliferation activity of EPCs was significantly increased in the VEGF165 treatment group （P〈0.05）, whilethe proliferation activity of EPCs was significantly inhibited and both the levels of proteins cyclin D 1 and PCNA were downregulatedin H20： treatment group （P 〈 0.05）. Compared with H2O2 treatment group, VEGF 165 pretreatment promoted EPCs proliferation significantly and caused increases of proteins cyclin D1 and PCNA expression （P〈0.05）. Conclusion VEGF165 may promote the proliferation of endothelial progenitor cells isolated from human peripheral blood and protect EPCs from H2O2-induced injury of proliferation activity because of the upregulation of proteins cyclin D1 and PCNA.

关 键 词：内皮祖细胞 过氧化氢 细胞增殖 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]