Fig 2: TCS induces pyroptosis of A549 cells. (A) Immunofluorescence determines the effect of different concentrations of TCS (0, 10, 20, and 40 µg/mL) on the expression of GSDMD-N. (B) Immunofluorescence determines the effect of different concentrations of TCS (0, 10, 20, and 40 µg/mL) on the ROS levels. (C) Western blotting measures the effect of different concentrations of TCS (0, 10, 20, and 40 µg/mL) on the pyroptosis-related proteins (NLRP3, ASC, caspase-1, pro caspase-1, GSDMD-N and GAPDH). *P<0.05, **P<0.01, and ***P<0.001 vs. 0 µg/mL. GSDMD, gasdermin-D; DAPI, 4',6-diamidino-2-phenylindole; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing a CARD; TCS, trichosanthin; ROS, reactive oxygen species.

Fig 3: Overexpression of NLRP3 reversed irisin-mediated antipyroptosis and anticardiac hypertrophy effects.A Immunoblots showing protein levels of NLRP3 and ASC (A1, A2), cleaved caspase-1, and GSDMD-N (A1, A3) in the suitably treated cardiomyocytes. B Percentage of PI-positive cells in the suitably treated cardiomyocytes. C Measurement of the cell surface area of cardiomyocytes in the indicated groups. D mRNA expression of cardiac hypertrophy markers BNP and ß-MHC in the suitably treated cardiomyocytes. Results are presented as the mean ± SEM, n = 6. *P < 0.01 compared to control, #P < 0.01 compared to Ang-II group, &P < 0.01 versus Ang-II + Irisin group.

Fig 4: Expression of NLRs, AIM2, and ASC associated with pyroptosis in TBI. (a) The immunoblot and (b–e) quantitative data of NLRs and AIM2 at 24 h post-CCI. Note that the increased expression of NLRs and AIM2 in the injured cortex after TBI was not inhibited by VX765 treatment. (f) The immunoblot and (g) quantitative data of ASC at 24 h post-CCI. The expression of ASC monomers was increased in the injured cortex and was not suppressed by VX765 treatment. In contrast, the increased ASC dimers and oligomers after TBI could be inhibited by VX765 treatment, suggesting that ASC oligomerization was blocked. (h) The immunostaining and (i) quantitative data of ASC in the injured cortex at 24 h post-CCI. As shown immunohistochemically, ASC was distributed in the damaged and surrounding areas. The cortex showed a strong increase in ASC staining in the CCI and DMSO groups compared to the VX765 group at 24 h post-TBI. (j) Co-IP examined the interactions between NLRP3 and ASC and proved that the interactions were significantly enhanced in the CCI and DMSO groups while VX765 treatment inhibited the binding between NLRP3 and ASC. Scale bars: 25 µm. n = 3. Data are presented as the mean ± SD. **P < 0.01, ***P < 0.001. N.S.: not significant.

Fig 5: rhKGF-2 has a role in regulating the activation of PI3K/Akt pathway, Nrf2/HO-1/NQO1 pathway and FoxO1-NLRP3-ASC-Caspase1 inflammasome in vivo. The SILI rat model was established by inhalation of smoke. Then, rhKGF-2 (10 ng/kg body weight), GSK2126458 (300 µg/kg body weight) and AS1842856 (10 µg/kg body weight) were used for were used for treating the SILI rats, the same volume of saline was administered on the rat in the sham group. Left lung tissues of rats in each group were collected for detection. A1-A2: WB was carried out to determine the expression of PI3K/Akt and FoxO1 in the lung tissues. B1-B2. The protein levels of Nrf2, HO-1 and NQO1 in the lung tissues. or nuclear were detected by WB. Nuclear Nrf2 was calculated using Histone H3 as internal control. C1-C2. WB was conducted to detect NLRP3-ASC-Caspase1 in the lung tissues. The relative protein expression (Y-axis) was calculated as fold change of sham group. Data are expressed as mean ± SD. All experiments were repeated for three times. Data difference was analyzed via ANOVA, and Student's t test. NSP>0.05, *p<0.05, **p<0.01, ***p<0.001. Five rats were involved in each group (N = 5).