Fig. 3

Pyroptosis in the murine model of TBI. a Expression of pyroptosis-related proteins in response to TBI was assessed by Western immunoblotting (WB). After 0, 1, 2, 4, 8, 12, and 24 h post-TBI, caspase-1, caspase-11, GSDMD, and caspase-1 protein fragments were severally elevated in WT-TBI mice compared to sham controls. All data are expressed as means ± SD, n = 5. Post-TBI at 24 h, caspase-1^−/− mice exhibited reduced TBI-induced pyroptosis. b In caspase-1^−/− mice, pyroptosis-related proteins were less prominent than the WT-TBI group, which demonstrated in WB results. All data are expressed as means ± SEM, n = 5. c Micrograph at × 20 magnification of caspase-1, caspase-11, and GSDMD immunostaining in the cortex in the vicinity of the lesion. As shown in the immunohistochemistry results, caspase-1, caspase-11, and GSDMD distributed in trauma organization and surrounding. The cortex showed a strong increase in caspase-1 and GSDMD staining in the WT-TBI compared to caspase-1^−/−-TBI at 24 h post-TBI. However, caspase-11 was no obvious difference between caspase-1^−/−-TBI and WT-TBI. One representative experiment of four was shown. d Caspase-1 activity was stimulated after TBI. In contrast, TBI-induced increase in caspase-1 activity was significantly reduced in caspase-1^−/− mice (###p < 0.001, WT-TBI vs. caspase-1^−/−-TBI). e Serum LDH was tested at 24 h following TBI since cerebral injury increases cellular LDH leakage (p < 0.001). Data are presented as the mean ± SEM. *p < 0.05 and ***p < 0.001 versus sham group, ###p < 0.001 versus TBI group, n = 6