An Oral Load of [13C3]Glycerol and Blood NMR Analysis Detect Fatty Acid Esterification, Pentose Phosphate Pathway, and Glycerol Metabolism through the Tricarboxylic Acid Cycle in Human Liver

J Biol Chem. 2016 Sep 2;291(36):19031-41. doi: 10.1074/jbc.M116.742262. Epub 2016 Jul 18.


Drugs and other interventions for high impact hepatic diseases often target biochemical pathways such as gluconeogenesis, lipogenesis, or the metabolic response to oxidative stress. However, traditional liver function tests do not provide quantitative data about these pathways. In this study, we developed a simple method to evaluate these processes by NMR analysis of plasma metabolites. Healthy subjects ingested [U-(13)C3]glycerol, and blood was drawn at multiple times. Each subject completed three visits under differing nutritional states. High resolution (13)C NMR spectra of plasma triacylglycerols and glucose provided new insights into a number of hepatic processes including fatty acid esterification, the pentose phosphate pathway, and gluconeogenesis through the tricarboxylic acid cycle. Fasting stimulated pentose phosphate pathway activity and metabolism of [U-(13)C3]glycerol in the tricarboxylic acid cycle prior to gluconeogenesis or glyceroneogenesis. Fatty acid esterification was transient in the fasted state but continuous under fed conditions. We conclude that a simple NMR analysis of blood metabolites provides an important biomarker of pentose phosphate pathway activity, triacylglycerol synthesis, and flux through anaplerotic pathways in mitochondria of human liver.

Keywords: anaplerosis; biomarker; gluconeogenesis; glycerol; liver metabolism; mitochondria; pentose phosphate pathway (PPP); tricarboxylic acid cycle (TCA cycle) (Krebs cycle); triglyceride.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Administration, Oral
  • Adult
  • Biomarkers / blood
  • Carbon Isotopes / administration & dosage
  • Carbon Isotopes / pharmacokinetics
  • Citric Acid Cycle / drug effects*
  • Esterification / drug effects
  • Esterification / physiology
  • Fatty Acids / blood*
  • Female
  • Glycerol / administration & dosage*
  • Humans
  • Liver / metabolism*
  • Magnetic Resonance Spectroscopy / methods*
  • Male
  • Middle Aged
  • Pentose Phosphate Pathway / drug effects*


  • Biomarkers
  • Carbon Isotopes
  • Fatty Acids
  • Glycerol