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Biochemistry/Physiology

Altered tricarboxylic acid cycle flux in primary myotubes from severely obese humans

International Journal of Obesity volume 43, pages 895–905 (2019)Cite this article

Subjects

Abstract

Background/objective

The partitioning of glucose toward glycolytic end products rather than glucose oxidation and glycogen storage is evident in skeletal muscle with severe obesity and type 2 diabetes. The purpose of the present study was to determine the possible mechanism by which severe obesity alters insulin-mediated glucose partitioning in human skeletal muscle.

Subjects/methods

Primary human skeletal muscle cells (HSkMC) were isolated from lean (BMI = 23.6 ± 2.6 kg/m2, n = 9) and severely obese (BMI = 48.8 ± 1.9 kg/m2, n = 8) female subjects. Glucose oxidation, glycogen synthesis, non-oxidized glycolysis, pyruvate oxidation, and targeted TCA cycle metabolomics were examined in differentiated myotubes under basal and insulin-stimulated conditions.

Results

Myotubes derived from severely obese subjects exhibited attenuated response of glycogen synthesis (20.3%; 95% CI [4.7, 28.8]; P = 0.017) and glucose oxidation (5.6%; 95% CI [0.3, 8.6]; P = 0.046) with a concomitant greater increase (23.8%; 95% CI [5.7, 47.8]; P = 0.004) in non-oxidized glycolytic end products with insulin stimulation in comparison to the lean group (34.2% [24.9, 45.1]; 13.1% [8.6, 16.4], and 2.9% [−4.1, 12.2], respectively). These obesity-related alterations in glucose partitioning appeared to be linked with reduced TCA cycle flux, as 2-[14C]-pyruvate oxidation (358.4 pmol/mg protein/min [303.7, 432.9] vs. lean 439.2 pmol/mg protein/min [393.6, 463.1]; P = 0.013) along with several TCA cycle intermediates, were suppressed in the skeletal muscle of severely obese individuals.

Conclusions

These data suggest that with severe obesity the partitioning of glucose toward anaerobic glycolysis in response to insulin is a resilient characteristic of human skeletal muscle. This altered glucose partitioning appeared to be due, at least in part, to a reduction in TCA cycle flux.

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Acknowledgements

We thank the research volunteers for their dedication and effort to this study. We also thank Gabe Dubis and Angela Clark for assisting in specimen collection. This study was supported by grants from Janssen Research & Development, LLC, Golden LEAF Foundation, National Institutes of Health (DK56112, JAH) and American Heart Association Postdoctoral Fellowship (15POST25080003, KZ). This publication was made possible by Mayo Clinic Metabolomics Resource Core through grant number U24DK100469 from the National Institute of Diabetes and Digestive and Kidney Diseases and originates from the National Institutes of Health Director’s Common Fund.

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Author notes

  1. J. Matthew Hinkley

    Present address: Translational Research Institute for Metabolism and Diabetes, Florida Hospital, Orlando, FL, USA

Authors and Affiliations

  1. Department of Exercise and Health Sciences, University of Massachusetts Boston, Boston, MA, USA

    Kai Zou

  2. Department of Kinesiology, East Carolina University, Greenville, NC, USA

    Kai Zou, J. Matthew Hinkley, Sanghee Park, Donghai Zheng & Joseph A. Houmard

  3. Human Performance Laboratory, East Carolina University, Greenville, NC, USA

    Kai Zou, J. Matthew Hinkley, Sanghee Park, Donghai Zheng & Joseph A. Houmard

  4. East Carolina Diabetes and Obesity Institute, East Carolina University, Greenville, NC, USA

    Kai Zou, J. Matthew Hinkley, Sanghee Park, Donghai Zheng, Walter J. Pories, G. Lynis Dohm & Joseph A. Houmard

  5. Department of Physical Therapy, East Carolina University, Greenville, NC, USA

    Terry E. Jones

  6. Department of Surgery, East Carolina University, Greenville, NC, USA

    Walter J. Pories

  7. Janssen Research & Development LLC, Spring House, PA, USA

    Pamela J. Hornby & James Lenhard

  8. Department of Physiology, East Carolina University, Greenville, NC, USA

    G. Lynis Dohm

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Correspondence to Kai Zou.

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Conflict of interest

P.J.H. and J.L. are employed by Johnson & Johnson. G.L.D. and W.J.P. received research grants from Janssen Research & Development, LLC.

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Zou, K., Hinkley, J.M., Park, S. et al. Altered tricarboxylic acid cycle flux in primary myotubes from severely obese humans. Int J Obes 43, 895–905 (2019). https://doi.org/10.1038/s41366-018-0137-7

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