Modulation of IκB kinase autophosphorylation and activity following brain ischemia  

脑缺血后IκB激酶的自身磷酸化及活性调节(英文)

作　　者：沈万华[1] 张春翌[2] 张光毅 

机构地区：[1]徐州医学院生物化学与分子生物学研究中心 [2]徐州医学院生物化学与分子生物学研究中心 徐州 221002 中国 [3]徐州 221002 中国

出　　处：《Acta Pharmacologica Sinica》2003年第4期25-29,96-97,共7页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China, № 30070182

摘　　要：AIM: To investigate the effects of different antagonists on the alteration of IκB kinase (IKK) activity in rat hippocampus following global brain ischemia. METHODS: Using 4-vessel occlusion (4-VO) as brain ischemia model, IKK protein expression was examined by immunoblotting and immunoprecipitation, and IKK activity was assayed by in vitro kinase assay. RESULTS: There was no alteration of IKK protein expression following ischemia or ischemia/reperfusion different time points, but IKK activity reached its peak level at ischemia 30 min. Pretreatment with N-methyl-D-aspartate (NMDA) receptor antagonist ketamine, non-NMDA receptor antagonist DNQX, or NF-κB inhibitor PDTC decreased the IKK activity following brain ischemia 30 min. The increase in substrate myelin basic protein (MBP) phosphorylation by IKK is associated with an increase in autophosphorylation of IKK, which can also be antagonized by ketamine, DNQX, and PDTC. CONCLUSION: NMDA receptor and non-NMDA receptor mediate the increase of IKK activity following global brain ischemia in rat hippocampus, which contributes to the alterations of expression and activity of downstream factor NF-κB.目的:观察脑缺血后IκB激酶的自身磷酸化及活性变化的调节机制。方法:采用IKKα/B抗体进行免疫印迹和免疫沉淀 分析,测定IKK蛋白表达,体外激酶活性分析测定IKK的自身磷酸化及活性。结果:IKK蛋白的表达在脑缺血不同时间以及脑缺血复灌不同时间都没有明显的变化;IKK的活性在缺血30分钟时达到最高;NMDA受体拮抗剂ketamine、非NMDA受体拮抗剂DNQX和NF-κB抑制剂PDTC均能抑制IKK的自身磷酸化及活性。结论:NMDA受体和非NMDA受体介导了脑缺血后IKK的自身磷酸化及活性调控,从而介导其下游因子NF-κB的蛋白表达及活性变化,参与脑缺血的损伤调控。

关 键 词：brain ischemia IκB kinase NF-κB HIPPOCAMPUS N-methyl-D-aspartate receptors reactive oxygen species signal transduction 

分 类 号：R743[医药卫生—神经病学与精神病学]