Fig 2: Bioinformatics analysis of the transcriptome sequencing results of cells. (A,B) Heatmap analysis of IRGs in cellular DEGs. (C–E) Expression levels of the pyroptosis proteins NLRP3 and ASC in SV-HUC-1 cells were detected by immunofluorescence staining (scale bar = 100 µm). (F) SEM showed TGF-ß1-induced epithelial cell swelling, cell membrane pore formation, and pyroptotic body generation. Scale bar = 5 µm. (G) TEM showed cell membrane disruption, disruption of organelle structures such as mitochondria and the endoplasmic reticulum, and vacuolization. Scale bar = 2 µm and scale bar = 500 nm. (H) Protein expression levels of NLRP3, GSDMD, ASC, and IL1-ß were detected by Western blotting in vitro. The data are presented as the mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Fig 3: The NF-?B signaling pathway and pyroptosis in rat bladder tissues after SCI. (A) Immunohistochemical staining of bladder tissue showed increased expression of P-P65 in the NB group (scale bar = 100 µm). (B) The ratio of the NF-?B pathway protein P-P65/P65 and IKBa expression levels in tissues were detected by Western blotting. (C) Serum levels of IL-6, IL-1ß, and IL-18 in rats (n = 5 each group). (D–F) Immunofluorescence staining and mean fluorescence intensity analysis of NLRP3 and ASC in bladder tissues (scale bar = 100 µm). (G) SEM showed disorganized cell arrangement in the epithelial layer of the rat bladder after SCI, increased filamentous secretions, and the generation of pyroptotic bodies. Scale bar = 10 µm, arrows indicate pyroptotic bodies. (H) TEM showed cell death, abnormal endoplasmic reticulum morphology and structural disruption. Scale bar = 2 µm, arrows indicate abnormal endoplasmic reticulum. (I) Protein expression levels of NLRP3, GSDMD, ASC, and IL1-ß in the two groups were detected by Western blotting. The data are presented as the mean ± SD; ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant.

Fig 4: Pyroptosis was involved in the process of bladder fibrosis in the bladder after SCI. (A) Serum levels of IL-1ß, IL-18 and LDH. (B) TUNEL staining of bladder tissue after SCI. (C) Western blot bands of NLRP3, GSDMD, IL-1ß and caspase1 and their relative optical density in bladder tissue at different time points after SCI. (D) Immunofluorescence of bladder sections showed upregulation of caspase1. (E) Immunofluorescence of bladder sections showed upregulation of NLRP3. Data are presented as the mean ± S.D. (n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001. Scale bars, 100 µm.