Inhibition on the S-nitrosylation of MKK4 can protect hippocampal CA1 neurons in rat cerebral ischemia/reperfusion.
S-nitrosylation, the nitric oxide-derived post-translational modification of proteins, plays critical roles in various physiological and pathological functions. In this present study, a rat model of cerebral ischemia and reperfusion by four-vessel occlusion was generated to assess MKK4 S-nitrosylation. Immunoprecipitation and immunoblotting were performed to evaluate MKK4 S-nitrosylation and phosphorylation. Neuronal loss was observed using histological detection. These results indicated that endogenous NO promoted the S-nitrosylation of MKK4. However, application of the exogenous NO donor S-nitrosoglutathione (GNSO), an inhibitor of the neuronal nitric oxide synthase 7-nitroindazole (7-NI), and the N-methyl-d-aspartate receptor (NMDAR) antagonist MK801 diminished I/R-induced S-nitrosylation and phosphorylation. These compounds also markedly decreased cerebral I/R-induced degeneration and death of neurons in hippocampal CA1 region in rats. Taken together, we demonstrated for the first time, that cerebral ischemia/reperfusion can induce S-nitrosylation of MKK4. We also found that inhibiting S-nitrosylation and activation of MKK4 resulted in marked decreases in neuronal degeneration and apoptosis, potentially via NMDAR-mediated mechanisms. These findings may lead to a new field of inquiry to investigate the underlying pathogenesis of stoke and the development of novel treatment strategies.
Research Center for Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, Xuzhou, 221002, PR China; Department of Laboratory Medicine, Affiliated Municipal Hospital of Xuzhou Medical College, Xuzhou, Jiangsu, 221002, PR China.
Xue Wen
XW
Wei
Research Center for Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, Xuzhou, 221002, PR China; Jiangsu Key Laboratory of Anesthesia and Analgesia Application Technology, Xuzhou, 221002, PR China.
Ling Yun
LY
Hao
Research Center for Biochemistry and Molecular Biology and Jiangsu Key Laboratory of Brain Disease Bioinformation, Xuzhou Medical College, Xuzhou, 221002, PR China. Electronic address: qisuhua9999@163.com.
Su Hua
SH
Qi
D000818:Animals; D002545:Brain Ischemia / Q000188:drug therapy / Q000473:pathology*; D056547:CA1 Region, Hippocampal / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism; D004195:Disease Models, Animal; D016291:Dizocilpine Maleate / Q000627:therapeutic use; D007191:Indazoles / Q000627:therapeutic use; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D008297:Male; D009474:Neurons / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D026422:S-Nitrosoglutathione / Q000627:therapeutic use*; D013997:Time Factors
D007191:Indazoles; D026422:S-Nitrosoglutathione; D016291:Dizocilpine Maleate; D048670:MAP Kinase Kinase 4; D053148:Caspase 3; C080122:7-nitroindazole
Cerebral ischemia and reperfusion (I/R); MKK4; NMDA receptor (NMDAR); Phosphorylation; S-nitrosylation