H_2O_2预处理对NF-κB的激活作用特征及其细胞保护作用  

Characteristics of effect of H_2O_2 preconditioning on NF-κB activation and its role in the cytoprotection in PC12 cells

作　　者：张梅[1] 郭瑞鲜[2] 莫利球[3] 崔宇[2] 刘微[2] 姚巧玲[2] 杨战利[2] 孟金兰[2] 陈培熹[2] 冯鉴强[2] 

机构地区：[1]广州医学院广州蛇毒研究所,广东广州510082 [2]中山大学中山医学院生理学教研室,广东广州510080 [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8期1023-1027,共5页Chinese Pharmacological Bulletin

基　　金：广东省科技计划资助项目(No2006B36004022)

摘　　要：目的探讨H2O2预处理对核转录因子-κB(NF-κB)的激活作用,并观察NF-κB在H2O2预处理诱导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诱导细胞凋亡的实验模型,分组如下:(1)空白对照组;(2)预处理组;(3)损伤组;(4)预处理+损伤组;(5)TPCK+预处理+损伤组;(6)TPCK组。应用碘化丙啶(PI)染色流式细胞术检测细胞凋亡率,甲氮甲唑蓝(MTT)法检测细胞存活率,免疫印迹法(Westernblot)测定NF-κB的表达水平,电泳迁移实验(EMSA)检测NF-κBDNA结合活性。结果H2O2预处理PC12细胞上调NF-κBp65的表达,增强DNA结合活性,并能明显地增加高浓度H2O2引起的NF-κBp65的表达(与损伤组比较,P<0.01)。H2O2预处理能使PC12细胞对抗高浓度H2O2引起的损伤,提高细胞存活率,降低细胞凋亡率(与损伤组比较,P<0.01)。NF-κB抑制剂甲苯磺酰苯丙氨酰氯甲酮(TPCK)可拮抗H2O2预处理对NF-κBp65的激活作用,并减弱H2O2预处理诱导的适应性细胞保护作用(与预处理+损伤组比较,P<0.01)。结论H2O2预处理对NF-κB的激活作用可能是其引起的适应性细胞保护机制之一。Aim To explore the effect of H2O2 precon- ditioning on nuclear factor - kappa B （ NF - KB ） activa -tion and the role NF-KB in cytoprotection induced by H2O2 preconditioning in PC12 cells. Methods The experimental model of H2O2 preconditioning against ap- optosis induced by H2O2 at a high concentration was established, and experimental groups were divided as follows ： （ 1 ） control; （2） precondition ; （ 3 ） H2O2 ; （ 4 ） precondition ＋ H2O2; （5） TPCK ＋ precondition ＋ H2O2 ; （ 6 ） TPCK. The percentage of apoptotic cells was measured by flow cytometry （FCM） with propidi- um iodide stain. The viability of PC12 cells was as- sessed by MTI＂ assay. The level of NF-KB expression was detected by Western blot. The NF-KB DNA bind- ing activity was detected by electrophoretic mobility shift assay （EMSA）. Results H202 preconditioning significantly up-regulated promoted its DNA binding preconditioning obviously NF-KB p65 expression and activity. Furthermore, H2O2 enhanced the expression of NF-KB p65 induced by H2O2 at a high dose （P 〈 0. 01 ,compared with H2O2 group）. H2O2 precondition- ing markedly protected PC12 cells against injury in- duced by H202 at a high dose, increasing the viability of cells and decreasing the percentage of apoptotic cells （P 〈 0. 01, compared with H2O2 group）. N-tosyl-L- phenylalanine chloromethyl ketone （TPCK） ,an inhibi- tor of NF-KB antagonized H2O2 preconditioning-in- duced NF-KB activation and the adaptive cytoprotection （P 〈 0. 01, compared with PC ＋ H2O2 group）. Conclusion NF-KB was activated by H202 preconditioning, which was one of the underlying mechanisms underlying adaptive cytoprotection induced by H2O2 precondi- tioning.

关 键 词：过氧化氢 预处理 核因子-ΚB 适应性细胞保护 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]