Research Paper Volume 15, Issue 20 pp 11184—11200

Figure 7. Effect of miRNA-200a-3p on H₂O₂-induced pyroptosis of HAECs via SIRT1. (A) qRT-PCR results showing the increased expression of miR-200a-3p in HAECs transduced with the miR-200a-3p mimics compared with negative control cells. (B, C) qRT-PCR and western blot results demonstrating the decreased SIRT1 RNA and protein expression in HAECs transduced with miR-200a-3p mimics. (D, E) qRT-PCR results showing the decreased SIRT1 RNA and protein expression in HAECs treated with H₂O₂ compared with cells that were not treated with H₂O₂. SIRT1 expression increased in HAECs transduced with the miR-200a-3p inhibitors compared with cells transduced with the NC inhibitor under H₂O₂ treatment. (F) Schematic of the complementary miR-200a-3p and SIRT1-3'UTR sequences. HEK-293T cells were co-transfected with wild-type or mutant SIRT1-3'UTR reporter constructs and miR-200a-3p mimics or corresponding negative controls (^**p<0.01 vs SIRT1-3'UTR-wt+NC mimics group, ns, not significant vs SIRT1-3'UTR-mut+NC mimics group)_. (G) Western blot demonstrating the decreased expression of SIRT1 in HAECs compared with that in negative control cells with or without H₂O₂ treatment. (H) Western blot demonstrating the decreased ratio of NLRP3, GSDMD-N, C-caspase-1, ASC, IL-1β, and IL-18 in cells transduced with miR-200a-3p inhibitor under H₂O₂ treatment compared with that in cells transduced with the NC inhibitor. The ratio of NLRP3, GSDMD-N, C-caspase-1, ASC, IL-1β, and IL-18 increased in cells transduced with the miR-200a-3p inhibitor and si-SIRT1 under H₂O₂ treatment compared with that in cells transduced with the miR-200a-3p inhibitor. (^*p<0.05 vs NC, ^**p<0.01 vs NC, ^***p<0.001 vs NC, ^#p<0.05 vs H₂O_2, ^##p<0.01 vs H₂O₂, ^&p<0.05 vs H₂O₂+Anti-miR-200a-3p, ^&&p<0.01 vs H₂O₂+ Anti-miR-200a-3p.). Data are presented as mean ± SD (n=3). HAECs, human aortic endothelial cells; qRT-PCR, quantitative real-time polymerase chain reaction; NC, negative control.