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Acute β-Hydroxy-β-Methyl Butyrate Suppresses Regulators of Mitochondrial Biogenesis and Lipid Oxidation While Increasing Lipid Content in Myotubes

  • Original Article
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Lipids

Abstract

Leucine modulates synthetic and degradative pathways in muscle, possibly providing metabolic benefits for both athletes and diseased populations. Leucine has become popular among athletes for improving performance and body composition, however little is known about the metabolic effects of the commonly consumed leucine-derived metabolite β-hydroxy-β-methyl butyrate (HMB). Our work measured the effects of HMB on metabolic protein expression, mitochondrial content and metabolism, as well as lipid content in skeletal muscle cells. Specifically, cultured C2C12 myotubes were treated with either a control or HMB ranging from 6.25 to 25 μM for 24 h and mRNA and/or protein expression, oxygen consumption, glucose uptake, and lipid content were measured. Contrary to leucine’s stimulatory effect on metabolism, HMB-treated cells exhibited significantly reduced regulators of lipid oxidation including peroxisome proliferator-activated receptor alpha (PPARα) and PPARβ/δ, as well as downstream target carnitine palmitoyl transferase, without alterations in glucose or palmitate oxidation. Furthermore, HMB significantly inhibited activation of the master regulator of energetics, AMP-activated protein kinase. As a result, HMB-treated cells also displayed reduced total mitochondrial content compared with true control or cells equivocally treated with leucine. Additionally, HMB treatment amplified markers of lipid biosynthesis (PPARγ and fatty acid synthase) as well as consistently promoted elevated total lipid content versus control cells. Collectively, our results demonstrate that HMB did not improve mitochondrial metabolism or content, and may promote elevated cellular lipid content possibly through heightened PPARγ expression. These observations suggest that HMB may be most beneficial for populations interested in stimulating anabolic cellular processes.

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Abbreviations

2-NBDG:

2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose

AKT:

Protein kinase B

AMPK:

AMP-activated protein kinase

BCAT2:

Branched-chain aminotransferase 2

BCKDH:

Branched-chain-alpha-keto acid dehydrogenase

CPT1b:

Carnitine palmitoyl transferase

FAS:

Fatty acid synthase

Foxo1:

Forkhead box protein O1

GLUT4:

Glucose transporter 4

MEF2:

Myocyte enhancer factor 2

mTOR:

Mammalian target of rapamycin

NRF1:

Nuclear respiratory factor 1

PGC-1α:

Peroxisome proliferator-activated receptor gamma coactivator 1 alpha

PPARα:

Peroxisome proliferator-activated receptor alpha

PPARβ/δ:

Peroxisome proliferator-activated receptor beta/delta

PPARγ:

Peroxisome proliferator-activated receptor gamma

SIRT1:

NAD+-dependent deacetylase sirtuin 1

SIRT3:

NAD+-dependent deacetylase sirtuin 3

TFAM:

Mitochondrial transcription factor A

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Acknowledgments

Support for this work was provided by the Department of Exercise Science within the Congdon School of Health Sciences. We would like also to thank the Department of Physical Therapy (Congdon School of Health Sciences) and the School of Arts and Sciences for the use of shared lab space and equipment. JKS performed experiments and was the lead author of the manuscript. MAJ and LMG assisted with experiments and manuscript preparation. NPG assisted with manuscript preparation. RAV conducted and oversaw all experiments, designed experiments, oversaw manuscript writing, performed statistical analyses, and conceived the study. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Department of Exercise Science, High Point University, One University Parkway, High Point, NC, 27268, USA

    Jamie K. Schnuck, Michele A. Johnson, Lacey M. Gould & Roger A. Vaughan

  2. School of Medicine, Medical College of Wisconsin, Milwaukee, WI, 53226, USA

    Nicholas P. Gannon

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  1. Jamie K. Schnuck
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  2. Michele A. Johnson
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Correspondence to Roger A. Vaughan.

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Schnuck, J.K., Johnson, M.A., Gould, L.M. et al. Acute β-Hydroxy-β-Methyl Butyrate Suppresses Regulators of Mitochondrial Biogenesis and Lipid Oxidation While Increasing Lipid Content in Myotubes. Lipids 51, 1127–1136 (2016). https://doi.org/10.1007/s11745-016-4193-2

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  • DOI: https://doi.org/10.1007/s11745-016-4193-2

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