Research Paper Volume 15, Issue 20 pp 11184—11200

Figure 2. Oxidative stress induces HAEC pyroptosis. (A) HAECs were treated with 0, 0.2, 0.4, or 0.6 mM H₂O₂ for 4 h, and the pyroptosis-related proteins were assessed via western blot analysis. H₂O₂-induced HAEC pyroptosis in a dose-dependent manner. (B) HAECs were treated with H₂O₂ (0.4 mM) for different durations (0 h, 2 h, 4 h, 8 h) and the pyroptosis-related proteins were assessed via western blot. H₂O₂ induced HAEC pyroptosis in a time-dependent manner. (C) Western blot revealing the downregulation of the NLRP3 protein in HAECs treated with siNLRP3 via transient transfection. (D) Western blot demonstrating the decreased protein levels of GSDMD-N, C-caspase-1, and IL-1β in HAECs transduced with siNLRP3 compared with cells transduced with NC under H₂O₂ treatment. (E) Western blot demonstrating the downregulation of the ASC protein in HAECs treated with siASC via transient transfection. (F) Western blot demonstrating the decreased protein levels of GSDMD-N and IL-1β in HAECs transduced with si-ASC compared with cells transduced with NC under H₂O₂ treatment. (G) Western blot demonstrating the decreased protein levels of C-caspase-1, GSDMD-N, and IL-1β in HAECs treated with VX-765 compared with cells without VX-765 under H₂O₂ treatment. Data are presented as mean ± SD (n=3). (^*p<0.05 vs control, H₂O₂ or H₂O₂+siNC, ^**p<0.01 vs control, H₂O₂ or H₂O₂+siNC, ^***p<0.001 vs control, ns, no significance). HAECs, human aortic endothelial cells; NC, negative control.