Fig. 8

Physical stress in mice induces the activation of pBTK–NLRP3–Caspase 1–IL1β pathway in the peripheral blood mononuclear cell (PBMC). Mice were subjected to physical stress by restraint and underwater trauma. A Immunoblot analysis of the PBMC homogenates using anti-NLRP3, cleaved Caspase 1, phospho-BTK (pBTK, Tyr223), BTK, and tubulin antibodies. B The relative densitometry of NLRP3 immunoblots of samples from the PBMC homogenates, which shows physical stress-induced higher levels of NLRP3 inflammasome in the PBMC of female mice in comparison to male mice. C Relative densitometry analysis of pBTK immunoblots showing physical stress significantly induced the activation of BTK, as revealed by increased pBTK levels in PBMC of stressed mice as compared to controls. D Caspase 1 activity in the PBMC homogenates using Caspase 1 assay kit: administration of ibrutinib in physically stressed mice showed reduced Caspase 1 activation, elucidating significant rescue of PBMC from the anxiogenic proinflammatory pathway. E Interleukin 1β (IL1β) levels in the plasma revealed physically stressed mice dosed with ibrutinib showed diminished plasma proinflammatory IL1β. All values are presented as mean with 95% CI (n = 10–14/group); ***p < 0.001, ns (not significant); 2X2X2 factorial ANOVA followed by Bonferroni post hoc test