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1,25(OH)2-vitamin D3 enhances the stimulating effect of leucine and insulin on protein synthesis rate through Akt/PKB and mTOR mediated pathways in murine C2C12 skeletal myotubes

Authors

  • Jérôme Salles,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
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  • Audrey Chanet,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
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  • Christophe Giraudet,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
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  • Véronique Patrac,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
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  • Philippe Pierre,

    1. Centre d'Immunologie de Marseille-Luminy, Aix-Marseille Université, Marseille, France
    2. Institut National de la Santé et de la Recherche Médicale, U1104, Marseille, France
    3. Centre National de la Recherche Scientifique, UMR 7280 Marseille, France
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  • Marion Jourdan,

    1. Nutricia Research, Utrecht, The Netherlands
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  • Yvette C. Luiking,

    1. Nutricia Research, Utrecht, The Netherlands
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  • Sjors Verlaan,

    1. Nutricia Research, Utrecht, The Netherlands
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  • Carole Migné,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
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  • Yves Boirie,

    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
    3. CHU Clermont-Ferrand, Service de Nutrition Clinique, Clermont-Ferrand, France
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  • Stéphane Walrand

    Corresponding author
    1. Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, Clermont-Ferrand, France
    2. INRA, UMR 1019, UNH, CRNH Auvergne, Clermont-Ferrand, France
    • Correspondence: Dr. Stéphane Walrand, Laboratoire de Nutrition Humaine, 58 rue Montalembert BP 321, 63009 Clermont-Ferrand Cedex 1, France

      E-mail: swalrand@clermont.inra.fr

      Fax: +33-4-73-60-82-55

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Abstract

Scope

In recent years, there has been a growing body of evidence pointing to an effect of vitamin D on muscle mass and function. Our aim was to investigate the combined effect of 1,25(OH)2-vitamin D3 (1,25(OH)2D3) with anabolic factors insulin and leucine on protein fractional synthesis rate (FSR) and regulation in the mouse C2C12 myotube.

Methods and results

After differentiation, myotubes were cultured in 1,25(OH)2D3 solutions at 0, 1, or 10 nM for 72 h. Cells were treated by l-[1–13C]valine and puromycin in presence or not of leucine and insulin, and protein FSR was determined by measuring tracer enrichments and puromycin incorporation in proteins, respectively. Protein expression and phosphorylation state of insulin receptor (IR), Akt, GSK3, mTOR, p70 S6 kinase, rpS6, and 4EBP1 were measured by Western blot. Transcript levels of IR and 1,25(OH)2D3 receptor (VDR) were determined by qPCR. 1,25(OH)2D3 (10 nM) with leucine and insulin increased protein FSR in C2C12 myotubes (14–16%). IR and VDR mRNA expression was increased with 1,25(OH)2D3 treatment. The Akt/mTOR-dependent pathway was activated by insulin and leucine and further enhanced by 1,25(OH)2D3.

Conclusion

1,25(OH)2D3 sensitizes the Akt/mTOR-dependant pathway to the stimulating effect of leucine and insulin, resulting in a further activation of protein synthesis in murine C2C12 skeletal myotubes.

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