p38丝裂原活化蛋白激酶对H2O2预处理的适应性细胞保护作用的影响  

Effect on adaptive cytoprotection of p38 mitogen-activated protein kinases for cells preconditioned by H2O2

作　　者：廖新学[1] 唐小卿[2] 郭瑞鲜[3] 陈正华[4] 杨春涛[3] 杨战利[3] 孟金兰[5] 陈培熹[3] 冯鉴强[3] 

机构地区：[1]中山大学附属第一医院心血管内科高血压血管病科,广州510080 [2]南华大学医学院生理学教研室 [3]中山大学中山医学院生理教研室 [4]中山大学中山眼科中心保健科 [5]广东药学院生理学教研室

出　　处：《中华生物医学工程杂志》2009年第3期203-207,共5页Chinese Journal of Biomedical Engineering

基　　金：广东省科技计划项目（2006B60501024）

摘　　要：目的探讨p38丝裂原活化蛋白激酶（MAPK）对H2O2预处理PC12细胞的适应性细胞保护作用的影响。方法在PC12细胞建立H2O2预处理对抗高浓度H2O2损伤的实验模型。将PC12细胞分组如下：（1）对照组，不加任何药物处理；（2）单纯H2O2处理组，细胞给予高浓度H2O2（50μmol／L）作用24h；（3）预处理组，10μmol／L H2O2作用PCI2细胞90min后撤去H2O2后继续常规培养24h；（4）预处理＋H2O2处理组，先用10μmol／L H2O2预处理，然后给予高浓度H2O2（50μmol／L）作用24h；（5）预处理＋H2O2处理＋p38抑制组，10μmol／L H2O2预处理后，在给以高浓度H2O2（50μmol／L）作用24h前20min加入10μmol／L p38特异性抑制剂SB203580；（6）p38抑制组，细胞仅用10μmol／L SB203580作用24h；（7）H2O2处理＋p38抑制组，PC12细胞在给以高浓度H2O2（50μmol／L）作用24h前20min加入10μmol／L p38特异性抑制剂SB203580。应用碘化吡啶（PI）染色，流式细胞术测定细胞凋亡率，免疫印迹法测定p38磷酸化水平并进行各组比较。结果单纯H2O2处理组细胞凋亡率较对照组明显升高[（48．33±7．41）％比（3．26±0．31）％，P〈0．01]。SB203580能显著抑制50μmol／L H2O2的致细胞凋亡作用，使细胞凋亡率降低[（23．42±3．52）％比（48．33±7．41）％，P〈0．01]。预处理＋H2O2处理组细胞凋亡率较单纯H2O2处理组明显下降[（16．93±3．50）％比（48．33±7．41）％，P〈0．01]。SB203580不影响H2O2预处理的适应性细胞保护作用。50μmol／L H2O2具有比H2O2预处理更强的激活p38的作用，H2O2预处理能明显地抑制50μmol／L H2O2对p38的激活作用。结论H2O2预处理抑制高浓度H2O2对p38的激活可能是其适应性细胞保护机制之一。Objective To investigate the effect on adaptive cytoprotection of p38 mitogen-activated protein kinases （MAPK） for PC12 cells preconditioned by H2O2. Methods Cytoprotection experimental model of PC12 cells was established, which was preconditioned by H2O2 against cellular injury induced by H2O2 at high concentration. All the PC 12 cells were devided into 7 groups： （1）control group： no treatment; （2） H2O2 simply treated group： PC 12 cells were treated only by 50 μmol/L H2O2 for 24 h; （3）preconditioning group： PC12 cells were pretreated with 10 μmol/L H2O2 for 90 min, followed by withdrawal of H2O2 and then routine culture for 24 h; （4）preconditioning＋H2O2 treated group： PC 12 cells were pretreated with 10 μmol/L H2O2, and subsequently exposed to 50 μmol/L H2O2 for 24 h; （5）preconditioning ＋ H2O2 treated ＋p38 inhibited group： PC12 eells were pretreated with 10 μ mol/L H2O2, followed by 10 μ mol/L SB203580 （speeifie inhibitor of p38） treatment for 20 min before exposure to 50 μmol/L H2O2 for 24 h; （6）p38 inhibited group： PC12 eells were only treated by 10 μmol/L SB203580 for 24 h; （7）H2O2 treated ＋ p38 inhibited group： PCI2 cells were treated by 10 μmol/L SB203580 for 20 rain before exposure to 50 μmol/L H2O2 for 24 h. Propidium iodide stain was performed, the apoptosis rate was determined by flow cytometry. The phosphorylation level of p38 was detected by Western blotting. The apoptosis rates and the phosphorylation levels of p38 were compared among different groups. Results The apoptosis rate of H2O2 simply treated group was obviously higher than that of control group [ （48.33±7.41）% vs （3.26±0.31）%, P〈 0.01]. SB203580 could inhibit apoptosis caused by 50 μmol/L H2O2, and the apoptosis rate decreased significantly [ （23.42 ± 3.52） % vs （48.33 ± 7.41 ） %, P〈0.01 ]. The apoptosis rate of preconditioning ＋ H2O2 treated group was obviously lower than that of H2O2 simply treated group E （16.93 ± 3.50）% vs （48.33 ± 7

关 键 词：过氧化氢 P38丝裂原活化蛋白激酶类 细胞凋亡 适应性细胞保护 预处理 

分 类 号：R285.5[医药卫生—中药学]