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The role of pyruvate dehydrogenase in mediating lipid-induced insulin resistance stands as a central question in the pathogenesis of type 2 diabetes mellitus. Many researchers have invoked the Randle hypothesis to explain the reduced glucose disposal in skeletal muscle by envisioning an elevated acetyl CoA pool arising from increased oxidation of fatty acids. Over the years, in vivo NMR studies have challenged that monolithic view. The advent of the dissolution dynamic nuclear polarization NMR technique and a unique type 2 diabetic rat model provides an opportunity to clarify. Dynamic nuclear polarization enhances dramatically the NMR signal sensitivity and allows the measurement of metabolic kinetics in vivo. Diabetic muscle has much lower pyruvate dehydrogenase activity than control muscle, as evidenced in the conversion of [1-13C]lactate and [2-13C]pyruvate to HCO3- and acetyl carnitine. The pyruvate dehydrogenase kinase inhibitor, dichloroacetate, restores rapidly the diabetic pyruvate dehydrogenase activity to control level. However, diabetic muscle has a much larger dynamic change in pyruvate dehydrogenase flux than control. The dichloroacetate-induced surge in pyruvate dehydrogenase activity produces a differential amount of acetyl carnitine but does not affect the tricarboxylic acid flux. Further studies can now proceed with the dynamic nuclear polarization approach and a unique rat model to interrogate closely the biochemical mechanism interfacing oxidative metabolism with insulin resistance and metabolic inflexibility.

Purpose: This study is to investigate time‐resolved 13C MR spectroscopy (MRS) as an alternative to imaging for assessing pyruvate metabolism using hyperpolarized (HP) [1‐13C]pyruvate in the human brain. Methods: Time‐resolved 13C spectra were acquired from four axial brain slices of healthy human participants (n = 4) after a bolus injection of HP [1‐13C]pyruvate. 13C MRS with low flip‐angle excitations and a multichannel 13C/1H dual‐frequency radiofrequency (RF) coil were exploited for reliable and unperturbed assessment of HP pyruvate metabolism. Slice‐wise areas under the curve (AUCs) of 13C‐metabolites were measured and kinetic analysis was performed to estimate the production rates of lactate and HCO3‐. Linear regression analysis between brain volumes and HP signals was performed. Region‐focused pyruvate metabolism was estimated using coil‐wise 13C reconstruction. Reproducibility of HP pyruvate exams was presented by performing two consecutive injections with a 45‐minutes interval. Results: [1‐13C]Lactate relative to the total 13C signal (tC) was 0.21–0.24 in all slices. [13C]HCO3‐/tC was 0.065–0.091. Apparent conversion rate constants from pyruvate to lactate and HCO3‐ were calculated as 0.014–0.018 s−1 and 0.0043–0.0056 s−1, respectively. Pyruvate/tC and lactate/tC were in moderate linear relationships with fractional gray matter volume within each slice. White matter presented poor linear regression fit with HP signals, and moderate correlations of the fractional cerebrospinal fluid volume with pyruvate/tC and lactate/tC were measured. Measured HP signals were comparable between two consecutive exams with HP [1‐13C]pyruvate. Conclusions: Dynamic MRS in combination with multichannel RF coils is an affordable and reliable alternative to imaging methods in investigating cerebral metabolism using HP [1‐13C]pyruvate. [ABSTRACT FROM AUTHOR]

Hyperpolarized 13C MRS allows the in vivo assessment of pyruvate dehydrogenase complex (PDC) flux, which converts pyruvate to acetyl-coenzyme A (acetyl-CoA). [1-13C]pyruvate has been used to measure changes in cardiac PDC flux, with demonstrated increase in 13C-bicarbonate production after dichloroacetate (DCA) administration. With [1-13C]pyruvate, the 13C label is released as 13CO2/13C-bicarbonate, and, hence, does not allow us to follow the fate of acetyl-CoA. Pyruvate labeled in the C2 position has been used to track the 13C label into the TCA (tricarboxylic acid) cycle and measure [5-13C]glutamate as well as study changes in [1-13C]acetylcarnitine with DCA and dobutamine. This work investigates changes in the metabolic fate of acetyl-CoA in response to metabolic interventions of DCA-induced increased PDC flux in the fed and fasted state, and increased cardiac workload with dobutamine in vivo in rat heart at two different pyruvate doses. DCA led to a modest increase in the 13C labeling of [5-13C]glutamate, and a considerable increase in [1-13C]acetylcarnitine and [1,3-13C]acetoacetate peaks. Dobutamine resulted in an increased labeling of [2-13C]lactate, [2-13C]alanine and [5-13C]glutamate. The change in glutamate with dobutamine was observed using a high pyruvate dose but not with a low dose. The relative changes in the different metabolic products provide information about the relationship between PDC-mediated oxidation of pyruvate and its subsequent incorporation into the TCA cycle compared with other metabolic pathways. Using a high dose of pyruvate may provide an improved ability to observe changes in glutamate. Copyright © 2013 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The interplay between glycolysis and gluconeogenesis is central to carbohydrate metabolism. Here, we describe novel methods to assess carbohydrate metabolism using [13C]‐probes derived from glycerate, a molecule whose metabolic fate in mammals remains underexplored. Isotope‐based studies were conducted via NMR and mass spectrometry analyses of freeze‐clamped liver tissue extracts after [2,3‐13C2]glycerate infusion. The ex vivo investigations were correlated with in vivo measurements using hyperpolarized [1‐13C]glycerate. Application of [13C]glycerate to N‐nitrosodiethylamine (DEN)‐treated rats provided further assessments of intermediary carbohydrate metabolism in hepatocellular carcinoma. This method afforded direct analyses of control versus DEN tissues, and altered ratios of 13C metabolic products as well as unique glycolysis intermediates were observed in the DEN liver/tumor. Isotopomer studies showed increased glycerate uptake and altered carbohydrate metabolism in the DEN rats. [ABSTRACT FROM AUTHOR]