Influence of calcium-induced workload transitions and fatty acid supply on myocardial substrate selection

Because of differences in energy yield and oxygen demand, the selection of oxidative fuels is important in the hypoxic or ischemic heart muscle. The aim of the present study was to clarify the contradictions observed in the effects of workload and fatty acid supply on myocardial fuel preference in isolated perfused rat hearts. Nuclear magnetic resonance spectroscopy combined with the administration of substrates labeled with the stable isotope carbon 13 and isotopomer analysis of glutamate labeling offers an opportunity to simultaneously measure metabolic fluxes in pathways feeding into the tricarboxylic acid (TCA) cycle. The work output was modulated by changes in extracellular calcium. In the presence of 5 mmol/L glucose, 0.5 mmol/L octanoate in the perfusate dominated the oxidative metabolism, and workload had little effect on the ratio of glucose to fatty acid utilization. This was the case even when the octanoate concentration was lowered to 50 micromol/L. The relative rate of replenishment of the TCA cycle intermediates was higher at a low workload. The redox state of flavoproteins in the intact heart was monitored fluorometrically to obtain an estimate of the mitochondrial reduced/oxidized nicotinamide-adenine dinucleotide ratio (NADH/NAD ratio) for assessment of the dominant level of regulation of cell respiration, and the myoglobin spectrum was simultaneously monitored to evaluate the oxygenation status of the myocardium. Commencement of octanoate infusion (50 micromol/L or 0.5 mmol/L) caused a large but transient reduction of mitochondrial NAD and, conversely, its cessation elicited NADH oxidation and rebound reduction. During glucose oxidation, an increase in workload led to oxidation of the mitochondrial NADH, but this effect was much smaller in the presence of 50 micromol/L octanoate and absent in the presence of 0.5 mmol/L. This indicates that strong control of oxygen consumption during glucose oxidation is exerted in the mitochondrial respiratory chain, whereas equal control during fatty acid oxidation is exerted within the metabolic pathway upstream from the respiratory chain. It is concluded that when a medium-chain fatty acid is available, myocardial workload and energy consumption have little influence on fuel preference and glucose oxidation remains suppressed.