【摘要】 目的:观察重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO)对肿瘤坏死因子-α(Tumor necrosis fac-tor-alpha,TNF-α)诱导的人脐静脉血内皮细胞(Human umbilical vein endothelial cells,HUVEC)凋亡的影响并探讨相关机制。方法:以不同浓度的rhEPO(12.5、25、50 IU/ml)预处理HUVEC 24 h,以50 ng/ml TNF-α诱导细胞凋亡,MTT法检测细胞生存率,ELISA法检测胞浆中细胞色素C(Cytochrome C,CytC)的浓度,免疫细胞化学法测定Caspase-3蛋白的表达,TUNEL法检测凋亡指数(Apoptosis index,AI),SCGE法(又称彗星实验)检测DNA损伤。结果:TNF-α作用HUVEC 24 h后细胞生存率和凋亡指数均高于正常对照组(P<0.05)。rhEPO以浓度依赖方式提高细胞生存率,降低CytC的浓度,抑制Caspase-3蛋白的表达,降低凋亡指数,减少DNA损伤(P均<0.05)。相关分析显示CytC的浓度与凋亡指数呈正相关(r=0.807,P<0.05);CytC的浓度与Cas-pase-3的相对含量呈正相关(r=0.583,P<0.05)。结论:rhEPO能够抑制TNF-α诱导的内皮细胞凋亡,可能与抑制线粒体凋亡通路的活化有关。更多还原

【Abstract】 Objective:To investigate the effects of recombinant human erythropoietin(rhEPO) on vascular endothelial cells apoptosis induced by tumor necrosis factor-alpha(TNF-α) and its possible mechanism.Methods:Human umbilical vein endothelial cells(HUVEC) were pretreated with rhEPO of different concentrations(12.5,25,50 IU/ml) before 50 ng/ml TNF-α treatment.The cell survival rate was measured by methyl thiazolyl tetrazolium(MTT) assay.The concentration of cytochrome C(CytC) in cytosol was analyzed using enzyme-linked immunosorbent assay(ELISA).The expression of Caspase-3 protein was examined by immunocytochemistry assay.The apoptosis index(AI) of HUVEC was detected by terminal dUTP nick-end 1abeling(TUNEL) method.The damage of DNA was evaluated using single cell gel electrophoresis(SCGE) assay.Results:rhEPO could significantly increase the cell survival rate,decrease the concentration of CytC,the relative content of Caspase-3,the AI of HUVEC and the damage of DNA in a concentration-dependent manner(all P<0.05).Correlation analysis showed that there was positive correlation between the concentration of CytC and AI(r=0.807,P<0.05).The concentration of CytC also had a positive correlation with the relative content of Caspase-3(r=0.583,P<0.05).Conclusion:rhEPO is a promising cell protector against apoptosis induced by TNF-α through interfering mitochondria-dependent apoptotic pathway.更多还原