Figure 8

NETs promoted NLRP3-mediated pyroptosis of HG-treated rat CCSMCs. (A) Assessment of the protein expression levels of NLRP3, GSDMD, GSDMD-N, pro-caspase-1, cleaved-caspase-1, and ASC in HG-treated CCSMCs using western blotting; the uncropped protein blots can be seen in Supplementary file 5. (B) Assessment of IL-1β and IL-18 levels in HG-treated CCSMCs using ELISA. sh-NC- or sh-NLRP3-transfected CCSMCs were subjected to HG (25 mM) treatment or/and co-cultured with 1 × 10⁴ neutrophils (N) for 24 h. CCSMCs cultured in 5 nM glucose served as controls. Data were presented as mean ± standard deviation. *p < 0.05, **p < 0.01, and ns denotes no significant difference between groups. NET neutrophil extracellular trap, NLRP3 NOD-like receptor family, pyrin domain-containing protein 3, HG high glucose, CCSMC corpus cavernosum smooth muscle cell, GSDMD gasdermin D, GSDMD-N N-terminal proteolytic fragment of gasdermin D, ASC apoptosis-associated speck-like protein containing a CARD, IL interleukin, ELISA enzyme-linked immunosorbent assay.