Fig. 8.

The KCa3.1 inhibitor TRAM-34 inhibits inflammasome activation in the colon and reduces DSS-induced colitis-like symptoms in mice at early time points. The mice were administered DSS and TRAM-34 as shown in Fig. 5. After 3 days or 5 days of DSS treatment, the mice were sacrificed, and proximal segments of the colon were collected for colon organ cultures, while middle and distal segments were subjected to H&E staining or immunofluorescence microscopy. A–C Weight loss (A), DAI (B) and colon lengths (C) of vehicle- or TRAM-34-treated mice on Day 3 or Day 5 of DSS exposure. D, E TRAM-34 administration attenuated DSS-induced colon injury. Representative images of colonic sections (H&E.) are shown (D). Scale bars, 100 µm. The total histological score of each section was calculated and is shown as a histogram (E). F, G TRAM-34 administration suppressed ASC speck formation in the DSS-exposed colon. Representative images of colonic ASC immunofluorescence from vehicle- or TRAM-34-treated mice are shown (F). Arrowheads indicate ASC specks. Scale bars, 20 µm. The number of ASC specks in each colon section was quantified (G). H, I TRAM-34 reduced the DSS-induced release of caspase-1p10 (Cas1p10) (H), IL-18 (H) and IL-1β (I) from colon organ cultures. The data are expressed as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001