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
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江苏省卫生厅基金资助项目(H200508)


Protective effects of eryrthropoietin on hypoxia/reoxygenation injury of neonatal SD rat cardiomyocytes
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    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2 h,复氧4 h;②EP组 在缺氧前1 h予EP(10 U/ml),随即缺氧2 h,复氧4 h;③Wortmannin+EP组(W+EP) 在EP预处理前30 min予3-磷脂酰肌醇激酶(phosphatidylinositol-3-kinase,PI3-K)特异性阻断剂Wortmannin(100 nmol/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。

    Abstract:

    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:①hypoxia/reoxygenation group(H/R):deoxygenation for 2 h and reoxygenation for 4 h;②EP group:EP(10 U/ml) was added 1 h before deoxygenation,then deoxygenation for 2 h and reoxygenation for 4 h;③Wortmannin+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;④Control 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.

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张锦英,张郁青,杨乃全,张定国.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[J].南京医科大学学报(自然科学版),2007,(5):428-431

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  • 收稿日期:2006-09-24
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