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, 8 (1), 798

Non-invasive Assessment of Hepatic Mitochondrial Metabolism by Positional Isotopomer NMR Tracer Analysis (PINTA)

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Non-invasive Assessment of Hepatic Mitochondrial Metabolism by Positional Isotopomer NMR Tracer Analysis (PINTA)

Rachel J Perry et al. Nat Commun.

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Abstract

Hepatic mitochondria play a central role in the regulation of intermediary metabolism and maintenance of normoglycemia, and there is great interest in assessing rates of hepatic mitochondrial citrate synthase flux (V CS) and pyruvate carboxylase flux (V PC) in vivo. Here, we show that a positional isotopomer NMR tracer analysis (PINTA) method can be used to non-invasively assess rates of V CS and V PC fluxes using a combined NMR/gas chromatography-mass spectrometry analysis of plasma following infusion of [3-13C]lactate and glucose tracer. PINTA measures V CS and V PC fluxes over a wide range of physiological conditions with minimal pyruvate cycling and detects increased hepatic V CS following treatment with a liver-targeted mitochondrial uncoupler. Finally, validation studies in humans demonstrate that the V PC/V CS ratio measured by PINTA is similar to that determined by in vivo NMR spectroscopy. This method will provide investigators with a relatively simple tool to non-invasively examine the role of altered hepatic mitochondrial metabolism.Liver mitochondrial metabolism plays an important role for glucose and lipid homeostasis and its alterations contribute to metabolic disorders, including fatty liver and diabetes. Here Perry et al. develop a method for the measurement of hepatic fluxes by using lactate and glucose tracers in combination with NMR spectroscopy.

Conflict of interest statement

The authors declare no competing financial interests.

Figures

Fig. 1
Fig. 1
VPC/VCS ratios measured by PINTA and ex vivo NMR. This ratio is identical whether measured by PINTA or ex vivo NMR analysis of rat livers
Fig. 2
Fig. 2
VPC/VEGP ratios measured by PINTA and ex vivo NMR. This ratio correlates tightly when measured by PINTA with the VPC/VEGP ratio determined using ex vivo NMR analysis
Fig. 3
Fig. 3
VPC/VCS ratios in healthy human control subjects. The measured ratios are comparable as measured by in vivo NMR at 4 T or by PINTA. Data are presented as the mean ± S.E.M. of n = 3
Fig. 4
Fig. 4
VPC/VEGP increases with an extended fast. The VPC/VEGP ratios measured using the two techniques are identical in both 6 and 48 h fasted rats. Data are the mean ± S.E.M. of n = 4 rats per time point

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