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The effect of oxygen in Sirt3-mediated myocardial protection: a proof-of-concept study in cultured cardiomyoblasts

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Abstract

Sirtuin 3 is a nicotinamide adenine dinucleotide dependent mitochondrial deacetylase that governs mitochondrial metabolism and oxidative defense. The demise in myocardial function following myocardial ischemia has been associated with mitochondrial dysfunction. Sirt3 maintains myocardial contractile function and protects from cardiac hypertrophy. The role of Sirt3 in ischemia is controversial. Our objective was to understand, under what circumstances Sirt3 is protective in different facets of ischemia, using an in vitro proof-of-concept approach based on simulated ischemia in cultured cardiomyoblasts. Cultured H9c2 cardiomyoblasts were subjected to hypoxia and/or serum deprivation, the combination of which we refer to as simulated ischemia. Apoptosis, as assessed by Annexin V staining in life-cell imaging and propidium-iodide inclusion in flow cytometry, was enhanced following simulated ischemia. Interestingly, serum deprivation was a stronger trigger of apoptosis than hypoxia. Knockdown of Sirt3 further increased apoptosis upon serum deprivation, whereas no such effect occurred upon additional hypoxia. Similarly, only upon serum deprivation but not upon simulated ischemia, silencing of Sirt3 led to a deterioration of mitochondrial function in extracellular flux analysis. In the absence of oxygen these Sirt3-dependent effects were abolished. These data indicate, that Sirt3-mediated myocardial protection is oxygen-dependent. Thus, mitochondrial respiration takes center-stage in Sirt3-dependent prevention of stress-induced myocardial damage.

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Funding

This work was funded by the National Health and Medical Research Council (NHMRC), the Victorian Government’s Operational Infrastructure Support Program, the Swiss National Science Foundation (310030, 146923), Matching Funds at the University of Zurich and the Zurich Heart House, Zurich, Switzerland. PD was supported by the German Research Foundation. JS was supported by the German Society of Cardiology. KP is a Principal Research Fellow of the NHMRC.

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Author notes

  1. Philipp Diehl, Daniel S. Gaul, as well as Karlheinz Peter and Stephan Winnik have contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology & Angiology I, Heart Center Freiburg University, Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Philipp Diehl, Jonas Sogl, Heiko Bugger, Christoph Bode, Martin Moser & Karlheinz Peter

  2. Baker IDI Heart and Diabetes Institute, Melbourne, Australia

    Philipp Diehl, Jonas Sogl, Ulrike Flierl, Darren Henstridge & Karlheinz Peter

  3. Department of Cardiology, University Heart Center Zurich, University Hospital Zurich, Ramistr. 100, 8098, Zurich, Switzerland

    Daniel S. Gaul, Frank Ruschitzka, Thomas F. Lüscher, Christian M. Matter & Stephan Winnik

  4. Hannover Medical School, Hanover, Germany

    Ulrike Flierl

  5. Center for Molecular Cardiology, University of Zurich, Zurich, Switzerland

    Juergen Pahla, Thomas F. Lüscher & Stephan Winnik

  6. Swiss Center for Regenerative Medicine, University of Zurich, Zurich, Switzerland

    Maximilian Y. Emmert & Christian M. Matter

  7. Zurich Center for Human Integrative Physiology (ZHIP), University of Zurich, Zurich, Switzerland

    Thomas F. Lüscher & Christian M. Matter

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  1. Philipp Diehl
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Correspondence to Stephan Winnik.

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Diehl, P., Gaul, D.S., Sogl, J. et al. The effect of oxygen in Sirt3-mediated myocardial protection: a proof-of-concept study in cultured cardiomyoblasts. J Thromb Thrombolysis 46, 102–112 (2018). https://doi.org/10.1007/s11239-018-1677-3

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