Integration of [U-13C]glucose and 2H2O for quantification of hepatic glucose production and gluconeogenesis |
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Authors: | Perdigoto Rui Rodrigues Tiago B Furtado Alexandre L Porto Armando Geraldes Carlos F G C Jones John G |
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Institution: | Medicine III, Transplantation Unit, University Hospital of Coimbra, 3049, Coimbra, Portugal. |
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Abstract: | Glucose metabolism in five healthy subjects fasted for 16 h was measured with a combination of U-13C]glucose and 2H2O tracers. Phenylbutyric acid was also provided to sample hepatic glutamine for the presence of 13C-isotopomers derived from the incorporation of U-13C]glucose products into the hepatic Krebs cycle. Glucose production (GP) was quantified by 13C NMR analysis of the monoacetone derivative of plasma glucose following a primed infusion of U-13C]glucose and provided reasonable estimates (1.90 +/- 0.19 mg/kg/min with a range of 1.60-2.15 mg/kg/min). The same derivative yielded measurements of plasma glucose 2H-enrichment from 2H2O by 2H NMR from which the contribution of glycogenolytic and gluconeogenic fluxes to GP was obtained (0.87 +/- 0.14 and 1.03 +/- 0.10 mg/kg/min, respectively). Hepatic glutamine 13C-isotopomers representing multiply-enriched oxaloacetate and U-13C]acetyl-CoA were identified as multiplets in the 13C NMR signals of the glutamine moiety of urinary phenylacetylglutamine, demonstrating entry of the U-13C]glucose tracer into both oxidative and anaplerotic pathways of the hepatic Krebs cycle. These isotopomers contributed 0.1-0.2% excess enrichment to carbons 2 and 3 and approximately 0.05% to carbon 4 of glutamine. |
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Keywords: | phenylacetylglutamine 2H NMR 13C NMR isotopomers gluconeogenesis Krebs cycle |
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