Activation and involvement of JNK1/2 in hydrogen peroxideinduced neurotoxicity in cultured rat cortical neurons  

作　　者：WeiWANG CanGAO Xiao-yuHOU YongLIU Yan-yanZONG Guang-yiZHANG 

机构地区：[1]ResearchCenterforBiochemistryandMolecularBiology,XuzhouMedicalCollege,Xuzhou221002,China [2]CorrespondencetoGuang-yiZHANG

出　　处：《Acta Pharmacologica Sinica》2004年第5期630-636,共7页中国药理学报（英文版）

基　　金：The present study was supported by grants from the Educa-tion Departmental Natural Science Research Funds of JiangsuProvince of China (No 99KJB180002; 99KJD180004) and theNational Natural Science Foundation of China (No 39770177).2

摘　　要：AIM: To investigate the role of c-Jun N-terminal protein kinase 1 and 2 (JNK1/2) and the main signal pathway forits activation in hydrogen peroxide (H2O2) induced apoptotic-like cortical cell death. METHODS: Using the modelof oxidative stress induced by H2O2, the expression and diphosphorylation of JNK1/2 was examined by immunoblottinganalysis, and neuronal apoptotic like cell death was determined by 4',6-diamidino-2-phenylindole (DAPI) staining.RESULTS: The elevation in diphosphorylation level of JNK1/2 (4.40-/5.61-fold vs sham control) was associatedwith the concentration of H2O2 (0-100 mol/L) and the development of apoptotic-like cell death (11.04 %-81.01 %).There was no alteration of JNK1/2 protein expression following H2O2 treatment and recovery at different timepoints. Administration with JNK1/2 antisense oligonucleotides not only significantly decreased JNK1/2 proteinexpression and activation level, but also significantly reduced cortical cell death induced by H2O2 exposure.Furthermore, both JNK1/2 diphosphorylation and apoptotic-like cell death were largely prevented by pretreatmentwith (5S,10R)-()-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801)or omission of Ca2+ in incubation medium with ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid(EGTA). CONCLUSION: JNK1/2 is activated and participates in H2O2-induced apoptotic-like death in cultured ratcortical neurons mainly via N-methyl-D-aspartate (NMDA) receptor-mediated influx of extracellular Ca2+.AIM: To investigate the role of c-Jun N-terminal protein kinase 1 and 2 (JNK1/2) and the main signal pathway forits activation in hydrogen peroxide (H2O2) induced apoptotic-like cortical cell death. METHODS: Using the modelof oxidative stress induced by H2O2, the expression and diphosphorylation of JNK1/2 was examined by immunoblottinganalysis, and neuronal apoptotic like cell death was determined by 4',6-diamidino-2-phenylindole (DAPI) staining.RESULTS: The elevation in diphosphorylation level of JNK1/2 (4.40-/5.61-fold vs sham control) was associatedwith the concentration of H2O2 (0-100 mol/L) and the development of apoptotic-like cell death (11.04 %-81.01 %).There was no alteration of JNK1/2 protein expression following H2O2 treatment and recovery at different timepoints. Administration with JNK1/2 antisense oligonucleotides not only significantly decreased JNK1/2 proteinexpression and activation level, but also significantly reduced cortical cell death induced by H2O2 exposure.Furthermore, both JNK1/2 diphosphorylation and apoptotic-like cell death were largely prevented by pretreatmentwith (5S,10R)-()-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801)or omission of Ca2+ in incubation medium with ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid(EGTA). CONCLUSION: JNK1/2 is activated and participates in H2O2-induced apoptotic-like death in cultured ratcortical neurons mainly via N-methyl-D-aspartate (NMDA) receptor-mediated influx of extracellular Ca2+.

关 键 词：c-Jun N-terminal protein kinase calcium N-methylaspartate receptors hydrogen peroxide SIGNALTRANSDUCTION apoptosis cerebral cortex 

分 类 号：Q55[生物学—生物化学] R741.02[医药卫生—神经病学与精神病学]