Hydrogen sulfide increases glutathione biosynthesis, and glucose uptake and utilisation in C₂C₁₂ mouse myotubes

Diabetic patients have lower blood concentrations of hydrogen sulfide (H₂S), L-cysteine (LC), and glutathione (GSH). Using C₂C₁₂ mouse myotubes as a model, this study investigates the hypothesis that the beneficial effects of LC supplementation are mediated by upregulation of the H₂S status under diabetic conditions. Results show that exogenous administration of sodium hydrosulfide (NaHS, 10 or 20 µM; 6 hours), a H₂S donor, significantly (p < .05) upregulates the gene expression of cystathionine-γ-lyase (CSE), LC transporter (Slc7a11/xCT), and the genes involved in GSH biosynthesis. Additionally, it reduces homocysteine (HCys), reactive oxygen species (ROS) production, and enhances cellular LC, H₂S, and glucose uptake and utilisation in myoblasts. The use of CSE siRNA to induce deficient endogenous H₂S production causes an increase in H₂O₂, ROS, HCys levels, and downregulation of GSH biosynthesis pathway enzymes. In additional, CSE knockdown downregulates glucose transporter type 4 (GLUT4) and gene expression of its key transcription factors, and reduces glucose uptake in C₂C₁₂ myotubes. CSE knockdown cells showed specific increases in the protein S-glutathionylation of LC transporter and GLUT4 along with increased total protein S-glutathionylation. Taken together, evidence from this study provides molecular insights into the importance of the CSE/H₂S system in maintaining the cellular glutathione and glucose homeostasis in C₂C₁₂ myotubes.