Research Paper Volume 15, Issue 20 pp 11184—11200

Figure 1. Oxidative stress induces HAEC injury. (A) The pyroptosis-related proteins were assessed via western blot in aortia in apoE^-/- mice. (B) qRT-PCR results showing the miRNA-200a-3p expression increased in HFD group compared with that in NCD group. (C) HAECs were treated with different concentrations of H₂O₂ (0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM) for 4 h, and cell viability was detected using the CCK-8 assay. (D) HAECs were treated with 0.4 mM H₂O₂ for different durations (0 h, 1 h, 2 h, 4 h, 8 h), and cell viability was detected using the CCK-8 assay. (E) LDH released from HAECs treated with H₂O₂ (0.4 mM) at different times. (F) H₂O₂ stimulation significantly increased intracellular ROS, which contributed to the induction of cell damage. Plotting scale =100 μm. (G) PI staining results revealed increased percentage of PI positive cells in H₂O₂-treated HAECs in a time-dependent manner. Double staining of PI (red) and Hoechst 33342 (blue). Plotting scale = 200 μm. (H) Morphological changes of HAECs under light microscope. Plotting scale = 200 μm. (*p<0.05 vs control, **p<0.01 vs control, ***p<0.001 vs control). Data are presented as mean ± SD (n=3). HAECs, human aortic endothelial cells; CCK-8, cell-counting kit 8; LDH, lactate dehydrogenase; ROS, reactive oxygen species; PI, propidium iodide; SD, standard deviation.