红细胞生成素抑制活性氧诱导的红细胞衰亡  

Erythropoietin inhibits eryptosis induced by reactive oxygen species

作　　者：孙云[1] 柳刚[1] 蒋雅丽[1] 张斌[1] 赵璇 李学刚[1] 

机构地区：[1]山东大学第二医院肾内科,山东济南250033

出　　处：《中国病理生理杂志》2017年第11期2084-2089,2094,共7页Chinese Journal of Pathophysiology

基　　金：山东大学第二医院种子基金资助项目(No.S2015010001);山东省自然科学基金资助项目(No.ZR2014HM045)

摘　　要：目的:观察红细胞生成素(erythropoietin,EPO)对过氧化氢(H_2O_2)刺激后红细胞衰亡(eryptosis)和红细胞中活性氧簇(reactive oxygen species,ROS)生成的影响,并探讨其可能机制。方法:将1%健康人红细胞悬液在以下3组不同的体外培养液中孵育:对照组(C组,培养基为PBS液)、H_2O_2组(H组,培养基为H_2O_2终浓度100μmol/L的PBS液)和EPO组(E组,培养基为H_2O_2终浓度100μmol/L、EPO终浓度2×10~4U/L的PBS液)。分别在孵育24 h和60 h时,留取红细胞以备检测。使用流式细胞术检测红细胞的衰亡率、红细胞内ROS和红细胞内钙离子浓度(_i^([Ca2+])),观察各检测指标的变化并分析其相关性。结果:红细胞衰亡率在C组随孵育时间延长而增加,在相同观察时点,H组较C组明显增加(P<0.01),E组较H组明显降低(P<0.01)。H组红细胞的ROS生成较C组明显增多,_i^([Ca2+])较C组明显升高(P<0.01);E组红细胞的ROS生成较H组明显减少,_i^([Ca2+])较H组明显降低(P<0.05或P<0.01)。结论:H_2O_2诱导健康红细胞加速衰亡,而EPO可以抑制H_2O_2诱导的红细胞衰亡,其机制可能与抗氧化及_i^([Ca2+])的改变有关。AIM：To observe the influence of erythropoietin（EPO）on eryptosis and production of reactive oxygen species（ROS）in erythrocytes under stimulation of hydrogen peroxide（H2O2）,.and to explore its related mechanism.METHODS：The erythrocyte suspension（1%）was cultured in vitro and divided into 3 groups：control group（C group,the culture medium was PBS）,H2O2group（H group,the culture medium was PBS containing H2O2at final concentration of 100μmol/L）and EPO group（E group,the culture medium was PBS containing H2O2at final concentration of 100μmol/L and EPO at final concentration of 2×104U/L）.The erythrocytes were collected at 24 h and 60 h.The eryptosis was detected by flow cytometry with Annexin V staining.The production of ROS and intracellular calcium ion concentration（[Ca2＋]i）were also analyzed by flow cytometry.RESULTS：The eryptosis in C group was increased as the incubating time extended.The eryptosis in H group was higher than that in C group（P〈0.01）,while that in E group was lower than that in H group（P〈0.01）.Meanwhile,ROS production and[Ca2＋]iwere higher in H group than those in C group（P〈0.01）,but those were lower in E group than those in H group（P〈0.05 or P〈0.01）.CONCLUSION：EPO inhibits eryptosis induced by H2O2and its mechanism may be related to antioxidant effect and change of[Ca2＋]i.

关 键 词：过氧化氢 红细胞衰亡 红细胞生成素 活性氧簇 细胞内钙离子浓度 

分 类 号：R363[医药卫生—病理学] R692.5[医药卫生—基础医学]