Fig. 1.

High-glucose and high-fat (HGHF) induces ferroptosis in human umbilical vein endothelial cells (HUVECs). HUVECs were treated with erastin, HGHF and ferrostatin-1 (Fer-1) for 24 h. (A) Cell viability of HUVECs exposed to different concentrations of palmitic acid (PA) for 24 h with or without HGHF (25.5 mM). (B) Cell viability of HUVECs exposed to different concentrations of Fer-1 for 24 h with or without HGHF. (C) Cell Counting Kit-8 (CCK-8) assay results of HUVECs pretreated with phosphate-buffered saline (PBS), 3-methyladenine (3-MA), Necrostatin-1 (Nec-1), Z-Val-Ala-Asp(OMe)-fluoromethylketone (Z-VAD-FMK), N-acetylcysteine (N-Ace), or Fer-1 followed by HGHF for 24 h. (D, E) Representative fluorescent images (D) and positive quantitative analysis (E) of HUVECs labeled with 5-ethynyl-20-deoxyuridine (EdU) (green); nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (F, G) Western blot (WB) analysis of solute carrier family 7 member 11 (SLC7A11), acyl-Coenzyme a synthetase long-chain family member 4 (ACSL4), and glutathione peroxidase 4 (GPX4) protein expression in differentially treated HUVECs: representative WB image (F) and quantitative analysis (G). (H, I) Representative fluorescent images (H) and quantitative analysis (I) of C11-BODIPY staining in differentially treated HUVECs. (J, K) WB analysis of SLC7A11, ACSL4, and GPX4 protein expression in wound tissues from the Control and diabetic wounds (DW) groups: representative WB image (J) and quantitative analysis (K). (L, M) Representative immunofluorescence images (L) and quantitative analysis (M) of GPX4 in wound tissues. Data are presented as the mean ± standard deviation (SD) from 5 independent experiments (n = 5). Statistical significance is denoted as ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001; ns indicates no significant difference.