It is known that nitric oxide (NO) influences the activities of glycolytic enzymes, resulting in alteration of glycolysis rate. We investigated the effect of NO donors on the energy metabolism of rat erythrocytes. Rat erythrocyterich blood suspensions were aerobically incubated for 2 hours (1) as controls or (2) with different concentrations of NO donors: nitroglycerin (NTG), isosorbide dinitrate (ISDN), molsidomine (MO), and sodium nitroprusside (SNP). NTG, ISDN, MO, and SNP significantly (p < 0.05) increased glucose consumption and lactate accumulation in rat erythrocytes in a dose-dependent manner, indicating stimulation of glycolysis. The increased rate of glycolysis was accompanied by elevation of energy production (p < 0.05), but no changes in ATP levels were observed. The dose-dependent increase of glycolytic ATP production and the unaltered levels of ATP resulted in considerably shortened ATP-turnover time with the maximal experimental doses of NO donors used, indicating the stimulation of the ATP-consuming process in rat erythrocytes. The metabolic effects of NTG, ISDN, MO, and SNP were not mimicked by exogenous 8-Br-cGMP, NaNO2, or NaNO3, suggesting that the NO donor-induced stimulation of glycolysis and abbreviation of ATP-turnover time in rat erythrocytes was mediated by NO as an effector molecule, irrespective of the irreversible scavenger effect of hemoglobin. The implications of the NO effect on energy metabolism of erythrocytes is discussed.