Fig. 9.

Impact of diosgenin (DG) application on diabetic wound (DW) healing and reactive oxygen species (ROS) diminishment in vivo. To evaluate the in vivo effects of DG on tube formation and ROS reduction, six groups were seted up: a non-diabetic control group (Control), a DW group (DW), and four diabetic treatment groups receiving either a low dose of DG (DW + DG^L), a high dose of DG (DW + DG^H), Fer-1 (DW + Fer-1), and a high dose of DG with OSS_128167 (DW + DG^H + OSS_128167). (A, B) Laser Doppler perfusion imaging showing angiogenesis recovery (A) and quantitative analysis (B). (C, D) Representative immunofluorescence images (C) and quantitative analysis (D) of α-smooth muscle actin (α-SMA) in wound from each group. (E, F) Representative immunofluorescence images (E) and quantitative analysis (F) of acyl-Coenzyme a synthetase long-chain family member 4 (ACSL4), in wound from each group. (G, H) Representative fluorescent images (G) and quantitative analysis (H) of 4-hydroxynonenal (4-HNE) in wound from each group. (I, J) Representative fluorescent images (I) and quantitative analysis (J) of Fe²⁺ in wound from each group. (K, L) Representative immunohistochemical images (K) and quantitative analysis (L) of Sirt6 in wound from each group. (M, N) Western blot (WB) (M) and quantitative analysis (N) of solute carrier family 7 member 11 (SLC7A11), ACSL4, and glutathione peroxidase 4 (GPX4) protein expression in wound tissues from the control groups, DW groups and DW + DG^H groups. Results are presented as the mean ± standard deviation (SD) from quintuple tests, n ≥ 5. Significance is denoted as ^∗P < 0.05, ^∗∗∗P < 0.001; ns indicates no significant difference. Fer-1: ferrostatin-1; OSS_128167: selective Sirt6 inhibitor.