Fig 2: PTUPB inhibits the activation of NLRP3 inflammasome in the lung of ALI mice. C57BL/6 mice received LPS injection (5 mg/kg, i.t.) with or without PTUPB pre-treatment (5 mg/kg) for 1 h. mRNA expression of Nlrp3 (A, n = 6-7), pro-caspase-1 (B, n = 6-7), Asc (C, n = 6-7), and pro-Il-1ß (D, n = 6-7) in the lungs was detected by RT-qPCR 12 h after the LPS injection. Protein expression of NLRP3 and pro-IL-1ß (E-G, n = 4) in the lungs was detected by Western blotting. mRNA expression of Nf-?b/p65 (H, n = 6-9) in the lungs was detected by RT-qPCR 12 h after the LPS injection. Protein expression of caspase-1p10 and IL-1ßp17 in the lungs was detected by Western blotting (I-K, n = 4). Concentration of IL-1ß in lung tissue was detected by ELISA (L, n = 5-8). Data are expressed as the mean ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Fig 3: Exosomes extracted from miR-326-overexpressed BMSCs significantly inhibited pyroptosis, HDAC3 expression, and STAT1/NF-?B p65 signaling pathway activation in OA chondrocytes, while silencing miR-326 had the opposite effect. (a) Exosomes extracted from miR-326-overexpressed BMSCs significantly promoted proliferation of OA chondrocytes, while silencing miR-326 had the opposite effect; (b) exosomes extracted from miR-326-overexpressed BMSCs significantly promoted migration of OA chondrocytes, while silencing miR-326 had the opposite effect; (c) exosomes extracted from miR-326-overexpressed BMSCs significantly increased the mRNA levels of chondrogenic specific genes (COL2A1, SOX9, Agg, and Prg4) in OA chondrocytes, while silencing miR-326 had the opposite effect; (d) exosomes extracted from miR-326-overexpressed BMSCs significantly decreased the levels of proinflammatory factors (IL-1ß, IL-18, IL-6, and TNF-a) in OA chondrocytes, while silencing miR-326 had the opposite effect; (e) exosomes extracted from miR-326-overexpressed BMSCs significantly decreased DNA-binding activity of NF-?B p65 in OA chondrocytes, while silencing miR-326 had the opposite effect; (f) exosomes extracted from miR-326-overexpressed BMSCs significantly decreased Caspase-1 activity in OA chondrocytes, while silencing miR-326 had the opposite effect; (g) immunofluorescence results showed that exosomes extracted from miR-326-overexpressed BMSCs significantly decreased the levels of GSDMD and cl-Caspase-1 in OA chondrocytes, while silencing miR-326 had the opposite effect; (h) Western blot results showed that exosomes extracted from miR-326-overexpressed BMSCs significantly decreased the level of HDAC3 and activated the STAT1/NF-?B p65 signaling pathway in OA chondrocytes, while silencing miR-326 had the opposite effect; (i) Western blot results showed that exosomes extracted from miR-326-overexpressed BMSCs significantly decreased the levels of pyroptosis-related proteins (NLRP3, ASC, GSDMD, Caspase-1, IL-1ß, and IL-18) in OA chondrocytes, while silencing miR-326 had the opposite effect; *P < 0.01, **P < 0.01.

Fig 4: Effects of AGM on inflammation-related proteins and mRNA in LPS-induced BV2 microglial cells. (A–E) Proteins expression level of inflammasome (NLRP3. ASC,CASPASE-1,IL-1ß) in different groups (Student’s test). (F–I) mRNA expression level of inflammasome (NLRP3,ASC,CASPASE-1,IL-1ß) in different groups (Student’s test). (J) The presence of IL-1ß was measured by ELISA (Student’s test). All data are shown as mean ± standard deviation.#p <0.05 vs the control and LPS group, *p<0.05 vs the LPS group and LPS + AGM group.

Fig 5: KO of Casp1/11 and Asc completely blocks FlaTox-induced cell death and mouse death.(A to E) Peritoneal macrophages were treated with FlaTox for 4 hours. LDH release (A), ATP loss (B), caspase-1 (C), caspase-8 (D), and caspase-3/7 activities (E) were measured (n = 3). (F) FlaTox-treated mice were monitored for survival (n = 8). (G) H&E staining of small intestinal tissue of mice treated for 3 hours. Scale bars, 100 µm. (H) WT mice were intraperitoneally injected with FlaTox, or FlaTox + belnacasan (100 mg/kg body weight) + emricasan (20 mg/kg body weight) and were monitored for survival (n = 6). (I to U) Peritoneal lavages from FlaTox-treated mice were isolated at 1 hour and 3 hours for caspase-1 (I), caspase-8 (J), caspase-3/7 (K), LDH (L), IL-1ß (M), TNF (N), and IL-6 (O) (n = 3). Peritoneal lavages at 20 min were collected for LC/MS/MS-based lipidomics (P and Q) (n = 3). Serum was used for LDH (R), IL-1ß (S), TNF (T), and IL-6 (U). n = 1 for PBS group and n = 3 for FlaTox-treated mice. (V) Model of death paths. (W and X) Mice were infected with Salmonella SL7207 and monitored for survival (n = 10). The bacterial loads in spleen and liver were analyzed at the time of mice death (n = 4). Graphs show means ± SE. OD490, optical density at 490 nm.