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Am J Physiol Endocrinol Metab 281: E848-E856, 2001;
0193-1849/01 $5.00
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Vol. 281, Issue 4, E848-E856, October 2001

An integrated 2H and 13C NMR study of gluconeogenesis and TCA cycle flux in humans

John G. Jones1, Michael A. Solomon2, Suzanne M. Cole3, A. Dean Sherry1,3, and Craig R. Malloy1,2

1 Department of Radiology, University of Texas Southwestern Medical Center, Dallas 75235; 2 Department of Internal Medicine, University of Texas Southwestern Medical Center and Department of Veterans Affairs Medical Center, Dallas 75216; and 3 Department of Chemistry, University of Dallas, Richardson, Texas 75083

Hepatic glucose synthesis from glycogen, glycerol, and the tricarboxylic acid (TCA) cycle was measured in five overnight-fasted subjects by 1H, 2H, and 13C NMR analysis of blood glucose, urinary acetaminophen glucuronide, and urinary phenylacetylglutamine after administration of [1,6-13C2]glucose, 2H2O, and [U-13C3]propionate. This combination of tracers allows three separate elements of hepatic glucose production (GP) to be probed simultaneously in a single study: 1) endogenous GP, 2) the contribution of glycogen, phosphoenolpyruvate (PEP), and glycerol to GP, and 3) flux through PEP carboxykinase, pyruvate recycling, and the TCA cycle. Isotope-dilution measurements of [1,6-13C2] glucose by 1H and 13C NMR indicated that GP in 16-h-fasted humans was 10.7 ± 0.9 µmol · kg-1 · min-1. 2H NMR spectra of monoacetone glucose (derived from plasma glucose) provided the relative 2H enrichment at glucose H-2, H-5, and H-6S, which, in turn, reflects the contribution of glycogen, PEP, and glycerol to total GP (5.5 ± 0.7, 4.8 ± 1.0, and 0.4 ± 0.3 µmol · kg-1 · min-1, respectively). Interestingly, 13C NMR isotopomer analysis of phenylacetylglutamine and acetaminophen glucuronide reported different values for PEP carboxykinase flux (68.8 ± 9.8 vs. 37.5 ± 7.9 µmol · kg-1 · min-1), PEP recycling flux (59.1 ± 9.8 vs. 27.8 ± 6.8 µmol · kg-1 · min-1), and TCA cycle flux (10.9 ± 1.4 vs. 5.4 ± 1.4 µmol · kg-1 · min-1). These differences may reflect zonation of propionate metabolism in the liver.

monoacetone glucose; acetaminophen glucuronide; carbon 13; deuterium; gluconeogenesis; liver metabolism


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