Fig. 4

Blocking Cx43-hemichannels in the UUO mouse provides anti-inflammatory protection by reducing NLRP3 inflammasome mediators and downstream drivers of inflammatory burden. Deletion of Cx43 from the proximal tubules protects against the UUO induced upregulation of NLRP3 and associated signalling mediators. To determine if this protection stems from a reduction in Cx43 hemichannel mediated signalling, we administered Peptide 5 via I.P. injections twice daily for 9 days, (9.3 mg/kg, 24 h after UUO) ahead of evaluating the impact that this has on NLRP3 and associated proteins. Immunostaining of the renal cortex and quantitative PCR (qPCR) analysis showed a marked downregulation in NLRP3 protein expression (a) and messenger RNA (mRNA) (b) in UUO mice treated with Peptide5 as compared with WT UUO. Similarly, blocking Cx43 hemichannels protected against an upregulation of IL1β (c), IL6 (d) and downstream NLRP3 inflammatory mediators, monocyte chemoattractant protein (MCP1) (e) and injury markers neutrophil gelatinase-associated lipocalin (N-GAL) (f) and vascular cell adhesion molecule (VCAM) (g). All groups are n = 4–6 with ANOVA and Tukey post-test used for experimental comparisons. *P < 0.05, **P < 0.01, and ***P < 0.001