Chemical versus isotopic equilibrium and the metabolic fate of glycolytic end products in the heart

Recent studies of isotope exchange across lactate dehydrogenase (LDH) and alanine aminotransferase (AAT) in hearts call into question whether both reactions are in equilibrium. To compare the oxidative and non-oxidative fates of glycolytic end products, isolated rabbit hearts were perfused with 5 mM [2-C-13] glucose and 2.5 mM [3-C-13] pyruvate: with (n = 6) and without (n = 7) stimulation of pyruvate oxidation using dichloroacetate (DCA), and during normal perfusion or hypoxia (n = 7/n = 6, + / - DCA). C-13 NMR spectroscopy of intact hearts confirmed a steady-state enrichment level in both alanine and lactate. H-1- and C-13-NMR spectroscopy of tissue extracts identified the fractions of lactate, alanine and glutamate pools formed from each exogenous substrate. Glycolysis from glucose accounted for 22 +/- 7% of lactate formed and 10 +/- 2% of alanine formed in control hearts, and 16 +/- 2% lactate and 15 +/- 2% alanine in hypoxic hearts (mean +/- S.E.M.). In contrast, exogenous pyruvate formed 36 +/- 5% of the lactate pool, and 86 +/- 3% of the alanine pool in controls and 47 +/- 3% of lactate and of 67 +/- 3% alanine during hypoxia. [2-C-13] glucose did not contribute to oxidative energy production via the TCA cycle as determined from low C-13 enrichment of glutamate C5 from glucose (<2%), while [3-C-13] pyruvate accounted for 84 +/- 7% of labeled glutamate C4. Thus, exogenous pyruvate out-competed the metabolism of glucose, indicating low glycolytic activity. At 40 min, 96 +/- 2% of the total alanine was labeled from either glucose or pyruvate, confirming equilibrium at AAT. However, only 55 +/- 10% of total lactate was labeled, suggesting that the LDH reaction is not in rapid equilibrium within the myocardium. (C) 1996 Academic Press Limited