Fig. 9

The effects of SAM and NaHS administration on NLRP3 inflammasome activation and MPO expression in P2X7R-overexpressing rats at 1 day after ICH. a, b Quantitative analysis of P2X7R and NLRP3 mRNA levels by qPCR in the vehicle, Ad-GFP, Ad-P2X7R, Ad-P2X7R + SAM and Ad-P2X7R + NaHS groups at 1 day after ICH. The mRNA levels of the P2X7R and NLRP3 were upregulated in Ad-P2X7R group (p < 0.05 vs. vehicle), but not in Ad-GFP group (p > 0.05 vs. vehicle), at 1 day after ICH. Administration of both SAM and NaHS inhibited the increase in P2X7R expression (p < 0.05 vs. Ad-P2X7R). The relative densities of each mRNA have been normalized against those of the vehicle group. c–e Representative bands and quantitative analysis of P2X7R, NLRP3 and mature IL-1β levels in the vehicle, Ad-GFP, Ad-P2X7R, Ad-P2X7R + SAM and Ad-P2X7R + NaHS groups at 1 day after ICH. The relative densities of each protein have been normalized against those of the vehicle group. f Representative bands and quantitative analysis of MPO levels in the vehicle, Ad-GFP, Ad-P2X7R, Ad-P2X7R + SAM and Ad-P2X7R + NaHS groups at 1 day after ICH. The relative densities of each protein have been normalized against those of the vehicle group. Western blotting performed at 1 day after ICH indicated that similar increases in the protein levels of the P2X7R and NLRP3 were induced by P2X7R overexpression and were suppressed by SAM and NaHS administration. Moreover, the protein levels of mature IL-1β and MPO were significantly elevated by overexpressing P2X7R at 1 day after ICH in the indicated group compared to the vehicle group (p < 0.05), but SAM or NaHS administration inhibited these increases (p < 0.05 vs. Ad-P2X7R). N = 5 rats per group. Data are presented as the mean ± SEM. #p < 0.05 vs. vehicle; $p < 0.05 vs. Ad-P2X7R. SAM S-adenosyl-l-methionine, NLRP3 pyrin domain-containing 3, GAPDH glyceraldehyde 3-phosphate dehydrogenase, ICH intracerebral haemorrhage, IL interleukin, Ad adenovirus, GFP green fluorescent protein, MPO myeloperoxidase