促红细胞生成素对乳鼠心肌细胞缺氧/复氧损伤的保护作用  

Protective effects of eryrthropoietin on hypoxia/reoxygenation injury of neonatal SD rat cardiomyocytes

作　　者：张锦英[1] 张郁青[2] 杨乃全[2] 张定国[2] 

机构地区：[1]南京医科大学第一附属医院肿瘤科,江苏南京210029 [2]南京医科大学第一附属医院心脏科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2007年第5期428-431,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生厅基金资助项目(H200508)

摘　　要：目的:观察促红细胞生成素(eryrthropoietin,EP)对乳鼠心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的影响及其相关机制探讨。方法:分离培养SD乳鼠心肌细胞,建立H/R模型。心肌细胞随机分为4组:①H/R组缺氧2h,复氧4h;②EP组在缺氧前1h予EP(10U/ml),随即缺氧2h,复氧4h;③Wortmannin+EP组(W+EP)在EP预处理前30min予3-磷脂酰肌醇激酶(phosphatidylinositol-3-kinase,PI3-K)特异性阻断剂Wortmannin(100nmol/L);④正常对照组流式细胞仪检测细胞凋亡率。Western blot法检测心肌细胞EP受体表达和p-AKt/AKt比值。结果:EP可明显降低缺氧/复氧引起的心肌细胞凋亡,并明显增强SD乳鼠心肌细胞p-AKt/AKt比值;而Wortmannin减弱了EP的抗细胞凋亡作用,显著降低了p-AKt/AKt比值。结论:细胞凋亡参与了心肌缺氧/复氧损伤;EP可减少缺氧/复氧引起的SD乳鼠心肌细胞凋亡,其机制可能与EP激活PI-3K/AKt信号通路有关。Objective：To examine the protective effects of eryrthropoietin （EP） on neonatal SD rat cardiomyocytes injury induced by hypoxia/reoxygenation and explore its mechanisms involved. Methods： The cultured neonatal SD rat cardiomyocytes were divided into four groups：（1）hypoxia/reoxygenation group（H/R）：deoxygenation for 2 h and reoxygenation for 4 h;（2）EP group：EP（10 U/ml） was added 1 h before deoxygenation,then deoxygenation for 2 h and reoxygenation for 4 h;（3）Wortmannin＋EP group（W＋EP group）： Wortmannin （100 nmol/L）, a specific phosphatidylinositol-3-kinase （PI3-K） inhibitor, was added at 30 min before EP incubation; （4）Control group. Cell apoptosis were measured by flow cytometry.The expression of EP receptor and p-Akt/Akt level were detected by Western blot. Results：EP significantly decreased the apoptotic index induced by hypoxia/reoxygenation and strongly increased p-AKt/Akt level. However, pretreatment with Wortmannin attenuated the cytoprotective effect of EP. Conclusion：Myocardial death and apoptosis are involved in hypoxia/reoxygenation injury,and EP can protect neonatal rat cardiomyocytes from hypoxia/ reoxygenation-induced apoptosis. The protective effects are partly associated with PI-3K/AKt pathway.

关 键 词：促红细胞生成素 心肌细胞 缺氧 复氧损伤 凋亡 蛋白激酶B 

分 类 号：Q74[生物学—分子生物学] R322.11[医药卫生—人体解剖和组织胚胎学]