Fig 2: POA protects PA-induced cell death via pyroptotic pathways.(A, B) Cells were treated with BSA, 200 μM PA, 200 μM PA+200 μM POA for four hours, and protein levels of cleaved caspase-11 (c-casp11), GSDMD-N, NLRP3, ASC, cleaved caspase-1 (c-casp1), cleaved IL-1β (c-IL 1β), GSDME-N were subsequently detected by Western blot. (C, D) Concentrations of IL-1β and IL-18 in cell culture supernatants were determined via ELISA. Data are shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

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: UCMSC-derived ApoEVs inhibited macrophage pyroptosis present during skin defect healing in db/db mice. a Immunofluorescence staining shows UCMSC-derived ApoEVs phagocytosed by F4/80-positive macrophages in vivo. Scale bar, 100 μm in low-magnification images, 20 μm in high-magnification images. b Tissue inflammatory factors IL-1β and IL-18 of different groups released in and around mouse skin defect area detected by ELISA, N = 3 per group, 3 duplication per sample. c Tissue protein levels of NLRP3, caspase-1, cleaved caspase-1, gasdermin D, cleaved gasdermin D of different groups released in and around mouse skin defect area detected by Western Blots, N = 3 per group. Corresponding uncropped full-length gels and blots are presented in Additional file 5B. d Semi-quantification of protein expression of NLRP3, cleaved caspase-1, and cleaved gasdermin D. e Immunofluorescence staining shows NLRP3, cleaved caspase-1, and gasdermin D in the skin defect area of mice in all three groups had significant co-localization with F4/80-positive macrophages. Scale bar, 40 μm in low-magnification images, 20 μm in high-magnification images. N = 3 per group, 3 ROIs per sample. f Semi-quantification of NLRP3, cleaved caspase-1, and gasdermin D relative fluorescence intensity in F4/80-positive cells of skin defect area. Data shown as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; NS not significant. CASP1 caspase-1; GSDMD, gasdermin D

Fig 5: NC009-1 down-regulated neuroinflammation and up-regulated cellular redox signaling in MPTP-treated mice. (A) Expression levels of striatal IL-1β, IL-6, and TNF-α analyzed by ELISA (n = 6). Expression levels of (B) NLRP3, CASP1, iNOS, and (C) SOD2, NRF2, and NQO1 in striatum analyzed by Western blot (n = 6). GAPDH was included as a loading control. To normalize, the relative NLRP3, CASP1, iNOS, SOD2, NRF2, and NQO1 of control mice was set as 100%. p values: comparisons between MPTP and control (# p < 0.05, ## p < 0.01, ### p < 0.001), or NC009-1-treated and untreated MPTP mice (* p < 0.05, ** p < 0.01).