Limited transfer of cytosolic NADH into mitochondria at high cardiac workload.
Glycolysis supplements energy synthesis at high cardiac workloads, producing not only ATP but also cytosolic NADH and pyruvate for oxidative ATP synthesis. Despite adequate Po(2), speculation exists that not all cytosolic NADH is oxidized by the mitochondria, leading to lactate production. In this study, we elucidate the mechanism for limited cytosolic NADH oxidation and increased lactate production at high workload despite adequate myocardial blood flow and oxygenation. Reducing equivalents from glycolysis enter mitochondria via exchange of mitochondrial alpha-ketoglutarate (alpha-KG) for cytosolic malate. This exchange was monitored at baseline and at high workloads by comparing (13)C enrichment between the products of alpha-KG oxidation (succinate) and alpha-KG efflux from mitochondria (glutamate). Under general anesthesia, a left thoracotomy was performed on 14 dogs and [2-(13)C]acetate was infused into the left anterior descending artery for 40 min. The rate-pressure product was 9,035 +/- 1,972 and 21,659 +/- 5,266 mmHg.beats.min(-1) (n = 7) at baseline (n = 7) and with dobutamine, respectively. (13)C enrichment of succinate was 57 +/- 10% at baseline and 45 +/- 13% at elevated workload (not significant), confirming oxidation of [2-(13)C]acetate. However, cytosolic glutamate enrichment, a marker of cytosolic NADH transfer to mitochondria, was dramatically reduced at high cardiac workload (11 +/- 1%) vs. baseline (50 +/- 14%, P < 0.05). This reduced exchange of (13)C from alpha-KG to cytosolic glutamate at high work indicates reduced shuttling of cytosolic reducing equivalents into the mitochondria. Myocardial tissue lactate increased 78%, countering this reduced oxidation of cytosolic NADH. The findings elucidate a contributing mechanism to glycolysis outpacing glucose oxidation in the absence of myocardial ischemia.
O'Donnell
J Michael
JM
Program in Integrative Cardiac Metabolism (MC 901), Dept. of Physiology and Biophysics, University of Illinois College of Medicine, 835 S. Wolcott Avenue, Chicago, IL 60612-7342, USA.
Kudej
Raymond K
RK
LaNoue
Kathyrn F
KF
Vatner
Stephen F
SF
Lewandowski
E Douglas
ED
D000085:Acetates / Q000493:pharmacokinetics; D000818:Animals; D002247:Carbon Isotopes; D003326:Coronary Circulation; D003600:Cytosol / Q000378:metabolism; D004285:Dogs; D004734:Energy Metabolism / Q000502:physiology*; D005260:Female; D018698:Glutamic Acid / Q000378:metabolism; D006321:Heart / Q000502:physiology*; D019344:Lactic Acid / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D009200:Myocardial Contraction / Q000502:physiology; D009206:Myocardium / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism
D000085:Acetates; D002247:Carbon Isotopes; D009243:NAD; D019344:Lactic Acid; D018698:Glutamic Acid; D019802:Succinic Acid; D010100:Oxygen