热休克蛋白参与PI3K/Akt介导H_2O_2预处理的抗凋亡作用  

Heat-shock proteins participate in H_2O_2 preconditioning-induced and PI3K/Akt signal pathway-mediated anti-apoptosis

作　　者：张梅[1] 李卫[2] 郭瑞鲜[3] 罗健东[1] 冯鉴强[3] 

机构地区：[1]广州医学院药理教研室,广东广州510082 [2]暨南大学附属第一医院神经外科,广东广州510632 [3]中山大学中山医学院生理教研室,广东广州510080

出　　处：《中国病理生理杂志》2011年第8期1557-1562,共6页Chinese Journal of Pathophysiology

基　　金：广州市属高校科技计划资助项目(No.10A178);广州医学院博士启动基金资助项目(No.2008C27)

摘　　要：目的:探讨热休克蛋白(HSP)是否参与PI3K/Akt信号通路介导的H2O2预处理的抗细胞凋亡作用。方法:利用PC12细胞建立H2O2预处理对抗高浓度H2O2诱导细胞凋亡的实验模型,分组如下:(1)空白对照组;(2)损伤组;(3)预处理+损伤组;(4)LY294002+预处理+损伤组;(5)LY294002组;(6)quercetin+预处理+损伤组;(7)17-AAG+预处理+损伤组;(8)溶剂对照组。应用Hoechst 33258染色观察细胞凋亡形态,碘化丙啶(PI)染色流式细胞术检测细胞凋亡率,比色法测定caspase-3的活性,免疫印迹法(Western blotting)测定HSP的表达水平。结果:100μmol/L H2O2预处理PC12细胞90 min可显著地抑制300μmol/L H2O2引起的细胞凋亡,使caspase-3的活性降低,同时上调HSP70和HSP90的表达。HSP70和HSP90的抑制剂quercetin和17-AAG拮抗H2O2预处理的抗细胞凋亡作用。PI3K抑制剂LY294002不仅拮抗了H2O2预处理抗细胞凋亡的作用,并且抑制H2O2预处理对HSP70和HSP90的表达上调。结论:PI3K/Akt通路、HSP70和HSP90均参与H2O2预处理诱导的细胞保护作用。并且HSP70和HSP90参与PI3K/Akt信号通路介导H2O2预处理的抗细胞凋亡作用。AIM ： To explore the effects of heat - shock proteins （HSP） on H2O2 preconditioning - induced and PI3 K/Akt pathway - mediated anti - apoptosis. METHODS ： The experimental model of H2O2 preconditioning against apoptosis （induced by H2O2 at a high concentration） was set up in cultured PC12 cells. The cells were divided into 8 groups as follows ： （ 1 ） control; （2） H2O2 ; （3） preconditioning ＋ H2O2 ; （4） LY294002 ＋ preconditioning ＋ H2O2 ; （5） LY294002 ; （6） quercetin ＋ preconditioning ＋ H2O2 ; （ 7 ） 17 - AAG ＋ preconditioning ＋ H2O2 ; （ 8 ） vehicle. Hoechst 33258 staining was used to observe the morphology of apoptotic changes. The percentage of apoptotic cells was estimated by flow cytometry with propidium iodide staining. The activity of caspase - 3 was assessed by colorimetry. The levels of HSP were detected by Western blotting assay. RESULTS： Preconditioning of H2O2 at the concentration of 100 μmol/L for 90 rain significantly protected PC12 cells from 300 μmol/L H2O2 - induced apoptosis, reduced the activity of caspase - 3, and upregulated the expression of HSP70 and HSP90. Quercetin and 17 -AAG （inhibitor of HSP70 and HSP90） antagonized anti - apoptosis induced by H2O2 preconditioning. LY294002 （ an inhibitor of PI3K） antagonized not only anti - apoptosis but also up- regulation of HSP70 and HSP90 induced by H2O2 preconditioning. CONCLUSION： The PI3K/Akt pathway, HSP70 and HSP90 involve in the cytoprotection induced by H2O2 preconditioning. Furthermore, HSP70 and HSP90 play a role in H2O2 preconditioninginduced anti -apoptosis mediated by PI3K/Akt pathway.

关 键 词：热休克蛋白质类 过氧化氢 预处理 PI3K/AKT通路 细胞凋亡 

分 类 号：R338.2[医药卫生—人体生理学]