Fig. 7

DCA intensifies TNBS-induced colitis by promoting IL-1β secretion in macrophages. a–e Littermate WT mice given or not given DCA in drinking water were treated with TNBS to induce experimental colitis, as described in Fig. 6a. On day 4, these mice were sacrificed, and their colon tissues were collected for the following analyses. a Relative mRNA levels of macrophages-associated cytokines (IL-1β, IL-6, TNF-α, and NOS2) and chemokines (CXCL1, CXCL2, CXCL10, and CCL7) were determined by real-time PCR and normalized to 18s. b Representative immunofluorescence images of F4/80 and IL-1β immunostaining in colon tissues (scale bars 200 μm). White boxes represent the magnified view. c The number of colonic F4/80⁺ IL-1β⁺ immune cells per 300 μm². Pyroptotic cells were co-immunostained with F4/80 (green) and IL-1β (red). F4/80⁺ IL-1β⁺ cells were counted under a microscope. d Pro-Caspase1, Cleaved-Caspase1, GSDMD-FL, GSDMD-NT, Pro-IL-1β, Cleaved-IL-1β immunoblot images in different treatment groups. e Quantitative analyses of relative protein levels in different treatment groups. The density of protein bands was quantified by Alpha-view software. a, c, and e Data represent means ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001; by unpaired Student’s t test. The data shown are representative of three independent experiments