ERK1／2介导H2O2预处理对抗氧化应激损伤的保护作用  

ERK 1/2 mediates the cytoprotection of H_2O_2 preconditioning against oxidative injury in PC 12 cells

作　　者：廖新学[1] 王艳丽[2] 郭瑞鲜[2] 张梅[2] 姚巧玲[2] 莫利求[3] 陈培熹[2] 冯鉴强[2] 

机构地区：[1]中山大学附属第一医院心血管内科 [2]中山大学中山医学院生理学教研室 [3]中山大学附属第一医院麻醉科,广东广州510080

出　　处：《中国药理学通报》2008年第9期1151-1156,共6页Chinese Pharmacological Bulletin

基　　金：广东省科技计划资助项目(No2006B60501024);广东省自然科学基金资助项目(No5001676)

摘　　要：目的探讨H2O2预处理能否激活ERK1/2及ERK1/2在H2O2预处理引起的适应性细胞保护中的作用。方法在PC12细胞,建立H2O2预处理对抗高浓度H2O2诱导细胞损伤的实验模型。应用甲氮甲唑蓝(MTT)法检测细胞存活率;碘化丙啶(PI)染色流式细胞术检测细胞凋亡率;免疫印迹法(Western blot)测定ERK1/2蛋白的表达及procaspase-3的表达。结果100μmol.L-1H2O2预处理PC12细胞90min能明显地保护PC12细胞对抗300μmol.L-1H2O2引起的损伤,使细胞存活率增加,细胞凋亡率降低及procaspase-3增多。H2O2预处理对ERK1/2具有明显的激活作用:诱导胞质ERK1/2磷酸化及促进其核转移。在H2O2预处理前30min应用ERK1/2抑制剂UO126(10μmol.L-1)可明显地阻断H2O2预处理的抗细胞毒性及抗细胞凋亡作用。结论H2O2预处理能激活ERK1/2,ERK1/2介导H2O2预处理的适应性细胞保护作用。Aim To explore whether H2O2 preconditioning can activate ERK 1 / 2 and the role of ERK 1 / 2 in H2O2 preconditioning-induced adaptive cytoprotection. Methods The experimental model of H2O2 preconditioning against PC 12 cells injury induced by H2O2 at high concentration was set up. The viability of cells was measured by MTT assay. The percentage of apoptotic cells was assessed by propidium iodide stain flow cytometry （FCM）. The levels of ERK1/2 and procaspase-3 were detected by Western blot assay. Results Preconditioning of H2O2 at 100 μmol.L^-1 for 90 min significantly protected PC 12 cells against 300 μmol.L^-1 H2O2-induced injury, increasing cell viability and procaspase-3 expression, and reducing per-cent of apoptotic cells.H2O2 preconditioning obviously induced cytosolic ERK1/2 phosphorylation and promo- ted its nuclear translocation. UO126 （ 10μmol.L^-1 ）, an inhibitor of ERK1/2,used 30 min before H2O2 preconditioning, markedly blocked the anti-cytotoxibity and anti-apoptosis induce by H2O2 preconditioning. Conclusion H2O2 preconditioning could activate ERK1/2 which mediates the adaptive cytoprotection of H2O2 preconditioning in PC 12 cells.

关 键 词：过氧化氢 预处理 ERK1/2 细胞保护 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R345.57[医药卫生—基础医学]