Research Paper Volume 15, Issue 20 pp 11184—11200

Inhibiting microRNA-200a-3p attenuates pyroptosis via targeting the SIRT1/NF-κB/NLRP3 pathway in H2O2-induced HAEC

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Figure 6. Effect of miRNA-200a-3p on H2O2-induced pyroptosis of HAECs. (A) HAECs were treated with H2O2 (0.4 mM) for 1 h, 2 h, 4 h, and 8 h; miRNA-200a-3p expression increased in all cases compared with that in control cells. (B) qRT-PCR results showing the decreased expression of miR-200a-3p in HAECs transduced with the miR-200a-3p inhibitor compared with negative control cells. (C) LDH release from HAECs treated with H2O2 and transduced with or without the miR-200a-3p inhibitor. (D) PI staining results showing the decreased percentage of PI positive cells in HAECs transduced with the miR-200a-3p inhibitor. (E) Western blot demonstrating increased ratio of NLRP3, GSDMD-N, C-caspase-1, ASC, IL-1β, IL-18 in cells treated with H2O2 compared with cells that were not treated with H2O2. The ratio of NLRP3, GSDMD-N, C-caspase-1, ASC, IL-1β, IL-18 decreased in cells transduced with the miR-200a-3p inhibitor under H2O2 treatment compared with cells transduced with the NC inhibitor. (*p<0.05 vs control, **p<0.01 vs control or NC, ***p<0.001 vs control or NC, #p<0.05 vs H2O2, ##p<0.01 vs H2O2). Data are presented as mean ± SD (n=3). HAECs, human aortic endothelial cells; qRT-PCR, quantitative real-time polymerase chain reaction; LDH, lactate dehydrogenase; PI, propidium iodide; NC, negative control.