Potential contribution of NF-kappaB in neuronal cell death in the glutathione peroxidase-1 knockout mouse in response to ischemia-reperfusion injury

Stroke; a Journal of Cerebral Circulation
Peter J CrackPaul J Hertzog

Abstract

We have previously identified an increased susceptibility of Gpx1(-/-) mice to increased infarct size after middle cerebral artery occlusion (MCAO). This study was designed to elucidate the mechanisms involved in elevated neuronal cell death arising from an altered endogenous oxidant state. Gpx1(-/-) mice were exposed to transient MCAO and reperfusion by intraluminal suture blockade. Protein expression of the p65 subunit of transcription factor nuclear factor-kappaB (NF-kappaB) was examined by immunohistochemistry and Western Analysis. NF-kappaB DNA-protein activity was assessed by electrophoretic mobility shift assays (EMSA). Wild-type and Gpx1(-/-) mice were exposed to MCAO with or without the NF-kappaB inhibitor, pyrrolidinedithiocarbamate (PDTC). Upregulation of the p65 subunit of NF-kappaB and subsequent p65 phosphorylation at serine 536 was detected in the Gpx1(-/-) brains after stroke. EMSAs revealed that increased ischemia-enhanced DNA binding of NF-kappaB was observed in Gpx1(-/-) mice compared with wild-type. Supershift assays indicated that the p50 and p65 subunits participated in the bound NF-kappaB complex. The NF-kappaB inhibitor PDTC, a potential antioxidant, was able to afford partial neuroprotection in the Gpx1...Continue Reading

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