An in vitro ischemic penumbral mimic perfusate increases NADPH oxidase-mediated superoxide production in cultured hippocampal neurons

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• 作者：Matthew E. Pamenter<sup>asup><sup> ; sup><sup>1sup><sup> ; sup><sup>mpamenter@ucsd.edusup> ; Sameh S. Ali<sup>bsup><sup> ; sup><sup>csup><sup> ; sup><sup>1sup> ; Qingbo Tang<sup>csup> ; J. Cameron Finley<sup>bsup> ; Xiang Q. Gu<sup>asup> ; Laura L. Dugan<sup>csup> ; Gabriel G. Haddad<sup>asup><sup> ; sup><sup>dsup><sup> ; sup><sup>esup>
• 关键词：AMPAR ; 2-amino-3-(5-methyl-3-oxo-1 ; 2-oxazol-4-yl)propanoic acid receptor ; Apo ; apocynin ; APV ; (2R)-amino-5-phosphonovaleric acid ; BBB ; blood brain barrier ; CNQX ; 6-cyano-7-nitroquinioxaline-2 ; 3-dione ; CrOx ; chromium oxalate ; DIDS ; 4 ; 4-diisothiocyanatostil
• 刊名：Brain Research
• 出版年：2012
• 出版时间：3 May, 2012
• 年：2012
• 卷：1452
• 期：Complete
• 页码：165-172
• 全文大小：827 K

文摘

The currently accepted scheme for reactive oxygen species production during ischemia/reperfusion injury is characterized by a deleterious mitochondria-derived burst of radical generation during reperfusion; however, recent examination of the penumbra suggests a central role for NADPH-oxidase (Nox)-mediated radical generation during the ischemic period. Therefore, we utilized a novel in vitro model of the penumbra to examine the free radical profile of ischemic murine hippocampal neurons using electron paramagnetic resonance spectroscopy, and also the role of Nox in this generation and in cell fate. We report that free radical production increased ~ 75 % at 2 h of ischemia, and this increase was abolished by: (1) scavenging of extracellular free radicals with superoxide dismutase (SOD), (2) a general anion channel antagonist, or (3) the Nox inhibitor apocynin. Similarly, at 24 h of ischemia, [ATP] decreased > 95 % and vital dye uptake increased 6-fold relative to controls; whereas apocynin, the Cl<sup>?/sup> channel antagonist 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), or the free radical scavenger N-acetyl cysteine (NAC) each provided moderate neuroprotection, ameliorating 13-32 % of [ATP]-depletion and 19-56 % of vital dye uptake at 24 h. Our results support a cytotoxic role for Nox-mediated free radical production from penumbral neurons during the ischemic period.