Fig. 4
Increase in neuronal COX-2 expression and prostaglandin E2 level in SHR rats following ischemia and reperfusion. (A-F) SD and SHR rats were subjected to sham surgery or MCAO (60 min) followed by reperfusion (RPF) for 6 h. (A-D) Tissue extracts from the ipsilateral cortex (MCAO-60’/RPF 6 h) were processed to measure mRNA levels of COX-2 by quantitative real-time-PCR. Transcript levels were normalized against housekeeping genes β-actin or HPRT. Bar graphs represent relative expression of COX-2 gene (mean ± SE, n = 5–6). (E) Coronal brain sections through the ipsilateral cortex of SD and SHR rats (MCAO-60’/RPF 6 h) were processed for immunohistochemical staining with anti-COX-2 antibody (red), anti-NeuN antibody (green) and nuclear stain DAPI (blue). Arrows in the representative photomicrographs demonstrate localization of COX-2 in NeuN positive cells in the ipsilateral cortex. (F) PGE2 level was measured by enzyme immunoassay in the supernatants from tissue lysates extracted from ipsilateral cortex of SD and SHR rats (MCAO-60’/RPF 6 h). Bar graphs represent quantitative analysis of PGE2 level (mean ± SD, n = 3). (A-D, F) Significant difference between means was assessed by student t-test and presented as *p < 0.01