Effect of hydrogen sulfide on glycolysis-based energy production in mouse erythrocytes

Hydrogen sulfide (H₂ S) is a gasotransmitter that regulates both physiological and pathophysiological processes in mammalian cells. Recent studies have demonstrated that H₂ S promotes aerobic energy production in the mitochondria in response to hypoxia, but its effect on anaerobic energy production has yet to be established. Glycolysis is the anaerobic process by which ATP is produced through the metabolism of glucose. Mammalian red blood cells (RBCs) extrude mitochondria and nucleus during erythropoiesis. These cells would serve as a unique model to observe the effect of H₂ S on glycolysis-mediated energy production. The purpose of this study was to determine the effect of H₂ S on glycolysis-mediated energy production in mitochondria-free mouse RBCs. Western blot analysis showed that the only H₂ S-generating enzyme expressed in mouse RBCs is 3-mercaptopyruvate sulfurtransferase (MST). Supplement of the substrate for MST stimulated, but the inhibition of the same suppressed, the endogenous production of H₂ S. Both exogenously administered H₂ S salt and MST-derived endogenous H₂ S stimulated glycolysis-mediated ATP production. The effect of NaHS on ATP levels was not affected by oxygenation status. On the contrary, hypoxia increased intracellular H₂ S levels and MST activity in mouse RBCs. The mitochondria-targeted H₂ S donor, AP39, did not affect ATP levels of mouse RBCs. NaHS at low concentrations (3-100 microM) increased ATP levels and decreased cell viability after 3 days of incubation in vitro. Higher NaHS concentrations (300-1000 microM) lowered ATP levels, but prolonged cell viability. H₂ S may offer a cytoprotective effect in mammalian RBCs to maintain oxygen-independent energy production.