Fig. 2
Inhibition of pyroptosis alleviated I/R and H/R injury in vivo and vitro. Kidneys and blood samples were collected from mice 24 hours after renal reperfusion following renal I/R. HK-2 cells were seeded at high density (125,000 cells/cm2, GJ formed). Cells in the hypoxia/reoxygenation (H/R) group were subjected to 24 hours of hypoxia followed by 4 hours of reoxygenation. A Representative H&E-stained kidney sections and kidney injury scores. Red arrow: cell swelling and cytoplasm rarefaction; Yellow arrow: tubular necrosis; Black arrow: cast formation; Green arrow: karyopyknosis; Blue arrow: inflammatory cells. Scale bar = 50 μM. * p < 0.05 vs. sham; # p < 0.05 vs. I/R. Data are means ± SEM (n = 5). B Serum samples were collected for measurements of serum creatinine and BUN levels. * p < 0.05 vs. sham; # p < 0.05 vs. I/R. Data are means ± SEM (n = 5). C Western blot analysis of NLRP3, GSDMD, and GSDMD-N expression using kidney lysates. * p < 0.05 vs. sham; # p < 0.05 vs. I/R mice. Data are means ± SEM (n = 3). D Concentration of IL-18 and IL-1β in serum analyzed by ELISA. * p < 0.05 vs. sham; # p < 0.05 vs. I/R. Data are means ± SEM (n = 5). E Morphology of HK-2 cells in different groups under scanning electron microscopy. Yellow arrow: bursting extensions. Scale bar = 10 μM. F Survival rate of HK-2 cells in different groups at high density. * p < 0.05 vs. control; # p < 0.05 vs. H/R. Data are means ± SEM (n = 5). G Western blot analysis of NLRP3, GSDMD, and GSDMD-N expression in HK-2 cells. * p < 0.05 vs. control; # p < 0.05 vs. H/R. Data are means ± SEM (n = 3). H Concentration of IL-18 and IL-1β in culture supernatant of HK-2 cells in different groups. * p < 0.05 vs. control; # p < 0.05 vs. H/R. Data are means ± SEM (n = 5)