NEUROPROTECTIVE EFFECTS OF PIOGLITAZONE IN A RAT MODEL OF PERMANENT FOCAL CEREBRAL ISCHEMIA ARE ASSOCIATED WITH PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR GAMMA-MEDIATED SUPPRESSION OF NUCLEAR FACTOR-κB SIGNALING PATHWAY

Previous studies have demonstrated that pioglitazone (Piog), a peroxisome proliferator-activated receptor gamma (PPAR gamma) agonist, inhibits ischemia-induced brain injury. Piog has also been shown to exert anti-inflammatory effects by attenuation of nuclear factor-kappa B (NF-kappa B) activation after myocardial ischemia/reperfusion injury. Because NF-kappa B is known to play a major role in the pathophysiology of brain ischemia, the present study was undertaken to elucidate whether pioglitazone attenuates ischemic neuronal damage through PPAR gamma-mediated suppression of NF-kappa B apoptotic signaling pathway. Permanent middle cerebral artery occlusion (pMCAO) model was induced by using an intraluminal filament technique in rats. Piog was administrated i.p. twice (24 h before and at the time of ischemia insult) or once (10 min after ischemia). The neuroprotection of Piog was analyzed by assessing neurological deficits, infarction volume and morphological changes. The inhibition of NF-kappa B signaling pathway by Piog was evaluated by detecting the nuclear translocation of NF-kappa B p65 with immunohistochemistry and its target gene p53 by real-time PCR, and the expression of phospholated NF-kappa B p65 (p- NF-kappa B p65) in primary cultured neurons and the protein levels of I kappa B alpha and p-ERK in the ischemic cortex or striatum with Western blotting analysis. The contribution of a PPAR gamma mechanism to Piog's inhibitory effects on NF-kappa B and neuroprotection was evaluated by pretreatment with the PPAR gamma irreversible antagonist GW9662. In vitro ischemia in cultured primary neurons was induced by the oxygen-glucose deprivation (OGD) and the protective effect of Piog on cultured neurons was measured by lactate dehydrogenase (LDH) assay. Piog (0.5, 1, 2 mg/kg) reduced infarction volume, and improved morphological changes and motor deficits. Piog markedly up-regulated the protein levels of I kappa B alpha or p-ERK 6 h or 12 h after ischemia. Piog reduced the nuclear translocation of NF-kappa B p65 in the ischemic cortical cells and repressed the expression of p53 12 h after ischemia. Pre-treatment with GW9662 blocked Piog-elicited reduction in infarction volume, the increase in protein levels of I kappa B alpha and p-ERK, the reduction in the nuclear translocation of NF-kappa B subunit p65 and the repression of p53 mRNA expression. In addition, Piog attenuated the OGD-induced neuronal damage and inhibited the OGD-induced increases in p- NF-kappa B p65 in neurons. The present findings suggest that Piog's neuroprotection appears to be associated with PPAR gamma-mediated suppression of NF-kappa B signaling pathway. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.