Activation and involvement of JNK1/2 in hydrogen peroxide-induced neurotoxicity in cultured rat cortical neurons.
OBJECTIVE
To investigate the role of c-Jun N-terminal protein kinase 1 and 2 (JNK1/2) and the main signal pathway for its activation in hydrogen peroxide (H(2)O(2)) induced apoptotic-like cortical cell death.


METHODS
Using the model of oxidative stress induced by H(2)O(2), the expression and diphosphorylation of JNK1/2 was examined by immunoblotting analysis, 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 associated with the concentration of H(2)O(2) (0-100 micromol/L) and the development of apoptotic-like cell death (11.04 %-81.01 %). There was no alteration of JNK1/2 protein expression following H(2)O(2) treatment and recovery at different time points. Administration with JNK1/2 antisense oligonucleotides not only significantly decreased JNK1/2 protein expression and activation level, but also significantly reduced cortical cell death induced by H(2)O(2) exposure. Furthermore, both JNK1/2 diphosphorylation and apoptotic-like cell death were largely prevented by pretreatment with (5S,10R)-(-)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801) or omission of Ca(2+) in incubation medium with ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA).


CONCLUSIONS
JNK1/2 is activated and participates in H(2)O(2)-induced apoptotic-like death in cultured rat cortical neurons mainly via N-methyl-D-aspartate (NMDA) receptor-mediated influx of extracellular Ca(2+).
Wang
Wei
W
Research Center for Biochemistry and Molecular Biology, Xuzhou Medical College, Xuzhou 221002, China.
Gao
Can
C
Hou
Xiao-yu
XY
Liu
Yong
Y
Zong
Yan-yan
YY
Zhang
Guang-yi
GY
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D016291:Dizocilpine Maleate / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000633:toxicity*; D048055:Mitogen-Activated Protein Kinase 8; D048056:Mitogen-Activated Protein Kinase 9; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D019579:Neocortex / Q000166:cytology* / Q000187:drug effects; D009474:Neurons / Q000166:cytology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors; D015398:Signal Transduction
D020319:Oligodeoxyribonucleotides, Antisense; D016194:Receptors, N-Methyl-D-Aspartate; D016291:Dizocilpine Maleate; D006861:Hydrogen Peroxide; D048056:Mitogen-Activated Protein Kinase 9; D048055:Mitogen-Activated Protein Kinase 8; D020928:Mitogen-Activated Protein Kinases