Tracer-based assessments of hepatic anaplerotic and TCA cycle flux: practicality, stoichiometry, and hidden assumptions

• Published in: American journal of physiology: endocrinology and metabolism Vol. 309; no. 8; pp. E727 - E735
• Main Authors: Previs, Stephen F., Kelley, David E.
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 15.10.2015
• Subjects: Acetaminophen - analogs & derivatives; Acetaminophen - metabolism; Acetates; Blood Glucose - metabolism; Carbon Isotopes; Citric Acid Cycle; Data interpretation; Fatty Liver - metabolism; Fluctuations; Glucose; Glutamic Acid - metabolism; Humans; Liver; Liver - metabolism; Metabolic Flux Analysis - methods; Metabolic Flux Analysis - standards; Metabolism; Mitochondria - metabolism; Mitochondria, Liver - metabolism; Oxidation-Reduction; Pathology; Plasma; Propionates; Radioactive Tracers; stable isotopes; fatty liver; NASH; metabolic disease; diabetes; insulin resistance
• DOI: 10.1152/ajpendo.00216.2015
• ISSN: 0193-1849; 1522-1555

Two groups recently used different tracer methods to quantify liver-specific flux rates. The studies had a similar goal, i.e., to characterize mitochondrial oxidative function. These efforts could have a direct impact on our ability to understand metabolic abnormalities that affect the pathophysiology of fatty liver and allow us to examine mechanisms surrounding potential therapeutic interventions. Briefly, one method couples the continuous infusion of [ 13 C]acetate with direct real-time measurements of [ 13 C]glutamate labeling in liver; the other method administers [ 13 C]propionate, in combination with other tracers, and subsequently measures the 13 C labeling of plasma glucose and/or acetaminophen-glucuronide. It appears that a controversy has arisen, since the respective methods yielded different estimates of the anaplerotic/TCA flux ratio (V ANA :V TCA ) in “control” subjects, i.e., the [ 13 C]acetate- and [ 13 C]propionate-derived V ANA :V TCA flux ratios appear to be ∼1.4 and ∼5, respectively. While the deep expertise in the respective groups makes it somewhat trivial for each to perform the tracer studies, the data interpretation is inherently difficult. The current perspective was undertaken to examine potential factors that could account for or contribute to the apparent differences. Attention was directed toward 1) matters of practicality, 2) issues surrounding stoichiometry, and 3) hidden assumptions. We believe that the [ 13 C]acetate method has certain weaknesses that limit its utility; in contrast, the [ 13 C]propionate method likely yields a more correct answer. We hope our discussion will help clarify the differences in the recent reports. Presumably this will be of interest to investigators who are considering tracer-based studies of liver metabolism.