Fig 2: AN1284 ameliorates hepatic steatosis and inflammation in db/db mice. The db/db mice exhibit increased liver weight (A), serum ALT (B), and ALP (C) levels as well as hepatic triglyceride and cholesterol contents (D,E). These parameters were prevented or ameliorated by AN1284 treatment. AN1284-treated mice also displayed decreased serum triglyceride levels (F), increased HDL/LDL cholesterol ratio (G), without an effect on total cholesterol levels (H). Quantification of hepatic fat content revealed an increase in db/db animals, which was normalized by AN1284 treatment (I). Representative liver images demonstrating macrovesicular steatosis in H&E-stained sections from db/db mice. Scale bar, 100 μm (J). The elevated hepatic mRNA expression levels of Acc (K), Scd1 (L), and Cd36 (M) in db/db mice were normalized by AN1284 treatment. Insignificant changes in hepatic TNFα protein levels were noted in non-treated and treated mice (N). Reduced protein expression of IL-18 (O), and MCP1 (P) in AN1284-treated db/db mice was documented. Scale bar, 50 μm. M1-to-M2 macrophage marker ratios displayed high content of M1 macrophages in db/db animals, which was diminished by AN1284 treatment (Q,R). Data represent the mean ± SEM from 8 to 10 mice per group. *P < 0.05 relative to control non-diabetic mice, #P < 0.05 relative to db/db-Veh-treated control mice.

Fig 3: Reproductive ageing is accompanied by an increase in the intra-ovarian expression of inflammatory cytokines. Gene expression levels of inflammasome-related genes Asc (A), Nlrp3 (B), Il18 (C) and Caspase1 (Casp1) (D) in ovaries from 2, 6, 12 and 18-month-old mice. n = 6 per cohort. Data are presented as mean ± SEM; (A,B,D): Ordinary One-way ANOVA, Tukey’s multiple comparisons test: a and b are significantly different among groups (p < 0.05). (C): Kruskal–Wallis test, Dunn’s multiple comparisons test: a and b are significantly different among groups (p < 0.05). (E) Representative image of a Western blot showing ovarian protein levels of ASC, NLRP3, IL-18 and cleaved Casp1 (cCASP1) from 2, 6, 12 and 18-month-old mice. Quantification of protein levels of ASC (F), NLRP3 (G), IL-18 (H) and cCASP1 (I) in ovaries from 2, 6, 12 and 18-month-old mice. n = 3–5 per cohort. Data are presented as mean ± SEM; E, F, Ordinary One-way ANOVA, Tukey’s multiple comparisons test (p > 0.05). (G,H); Kruskal–Wallis test, Dunn’s multiple comparisons test (p > 0.05). Each dot represents one animal.

Fig 4: The inhibition of miR-92a-3p alleviated TEC pyroptosis in I/R-induced kidney of mice. (A) Representative photomicrographs of tubular cell injury in mouse kidney tissue sections with HE staining, TUNEL staining, and representative photomicrographs of IL-1β and IL-18 expression in mouse kidney tissue sections by immunohistochemistry, 400×, scale bar = 20 μm. (B) Statistical quantification analysis showed the injury score of HE staining in the kidney tissues. (C) Statistical analysis showed the percentage of TUNEL-positive TECs in the kidney tissues. (D) Statistical analysis showed the positive area of IL-1β and IL-18 in the kidney tissues. (E,F) Western blot analysis of Nrf1, GSDMD-N, NLRP3, and caspase-1 expression in mouse kidney tissue sections. SCr levels (G) and BUN levels (H) were detected in mice. Data are expressed as the mean ± SD. n = 5 per group. *P < 0.05 vs. antagomir NC sham group. ∧P > 0.05 vs. antagomir NC sham group. #P < 0.05 vs. I/R-induced antagomir NC group, one-way ANOVA.

Fig 5: Knockdown of WHSC1 attenuated LPS-induced pyroptosis. (a) The protein levels of NLRP3, ASC, caspase-1 p20, IL-18, IL-1β, GSDMD FL and GSDMD CL in alveolar macrophages of mice introduced into Ad-shWHSC1 and treated with 10 mg/kg LPS were detected using Western blot. (b) The activity of caspase-1 in the alveolar macrophage of mice introduced into Ad-shWHSC1 and treated with 10 mg/kg LPS was determined. Ns: P > 0.05.