Fig. 2.

Role of diosgenin (DG) in counteracting high-glucose and high-fat (HGHF)-induced ferroptosis in human umbilical vein endothelial cells (HUVECs). To validate the role of DG in ferroptosis, five groups were set up: control, HGHF, HGHF + low dose of DG (10 μM) (DG^L), HGHF + high dose of DG (20 μM) (DG^H), and HGHF + ferrostatin-1 (Fer-1). (A) Chemical structure of DG. (B, C) Cell viability percentages after exposure to different concentrations of DG for 24 h (B) and 48 h (C). (D, E) Cell viability percentages after pretreatment with HGHF (D) and erastin (E), combined with or without different doses of DG. (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) Representative images of mitochondrial morphology in differentially treated HUVECs. (I, J) Representative fluorescent images (I) and quantitative analysis (J) of C11-BODIPY staining in differentially treated HUVECs. (K, L) Representative fluorescent images (K) and quantitative analysis (L) of FerroOrange staining in differentially treated HUVECs. 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.