Alpha-ketoglutarate enhances adipose-derived stem cells survival in wound healing by hypoxia-inducible factor 1-alpha-mediated redox homeostasis and glycogen-dependent bioenergetics

Figure 5 The combinational changes in glutamine and glycogen metabolism are necessary for cell viability in α-ketoglutarate-adipose-derived stem cells. A and B: Analysis of cell viability of adipose-derived stem cells (ADSCs), α-ketoglutarate-ADSCs (αKG-ADSCs), and αKG-ADSCs treated with BPTES (A) or 1,4-dideoxy-1,4-imino-d-arabinitol (DAB) (B) during hypoxia (0.5% O₂); C-F: Intracellular reactive oxygen species levels (C and E) and quantification of intracellular ATP levels (D and F) in ADSCs, αKG-ADSCs, and αKG-ADSCs treated with BPTES (C and D) or DAB (E and F) during hypoxia (0.5% O₂); G: Cell viability of ADSCs, αKG-ADSCs, and αKG-ADSCs treated with BPTES and DAB during normal or combined stress conditions (1% O₂), 1 mmol/L glucose, and 12 mmol/L H₂O₂ (high H₂O₂); H: Cell viability of αKG-ADSCs in 3 days after in vivo ectopic ADSCs implantation. Viable cells are CM-FDA⁺AnxV-PI^-; I and J: Terminal deoxynucleotidyl transferase dUTP nick end labeling immunostaining (I) at day 3 after cell implantation, with quantification of the percentage of TUNEL+ cells (J); K: Ratio of glutathione to glutathione disulfide in implanted ADSCs, αKG-ADSCs, and GLS1^KDPYGL^KD αKG-ADSCs; L: In vivo intracellular reactive oxygen species levels in implanted ADSCs, αKG-ADSCs, and GLS1^KDPYGL^KD αKG-ADSCs; M: Glycogen content in implanted ADSCs, αKG-ADSCs, and GLS1^KDPYGL^KD αKG-ADSCs. Data are presented with means ± SEM (^aP < 0.05, ^bP < 0.01). ADSCs: Adipose-derived stem cells; αKG-ADSCs: Alpha-ketoglutarate adipose-derived stem cells; GSH: Glutathione; GSSG: Glutathione disulfide; MFI: Mean fluorescent intensity; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling.