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TNFα protects cardiac mitochondria independently of its cell surface receptors

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Abstract

Our novel proposal is that TNFα exerts a direct effect on mitochondrial respiratory function in the heart, independently of its cell surface receptors. TNFα-induced cardioprotection is known to involve reactive oxygen species (ROS) and sphingolipids. We therefore further propose that this direct mitochondrial effect is mediated via ROS and sphingolipids. The protective concentration of TNFα (0.5 ng/ml) was added to isolated heart mitochondria from black 6 × 129 mice (WT) and double TNF receptor knockout mice (TNFR1&2−/−). Respiratory parameters and inner mitochondrial membrane potential were analyzed in the presence/absence of two antioxidants, N-acetyl-l-cysteine or N-tert-butyl-α-(2-sulfophenyl)nitrone or two antagonists of the sphingolipid pathway, N-oleoylethanolamine (NOE) or imipramine. In WT, TNFα reduced State 3 respiration from 279.3 ± 3 to 119.3 ± 2 (nmol O2/mg protein/min), increased proton leak from 15.7 ± 0.6% (control) to 36.6 ± 4.4%, and decreased membrane potential by 20.5 ± 3.1% compared to control groups. In TNFR1&2−/− mice, TNFα reduced State 3 respiration from 205.2 ± 4 to 75.7 ± 1 (p < 0.05 vs. respective control). In WT mice, both antioxidants added with TNFα restored State 3 respiration to 269.2 ± 2 and 257.6 ± 2, respectively. Imipramine and NOE also restored State 3 respiration to 248.4 ± 2 and 249.0 ± 2, respectively (p < 0.01 vs. TNFα alone). Similarly, both antioxidant and inhibitors of the sphingolipid pathway restored the proton leak to pre-TNF values. TNFα-treated mitochondria or isolated cardiac muscle fibers showed an increase in respiration after anoxia–reoxygenation, but this effect was lost in the presence of an antioxidant or NOE. Similar data were obtained in TNFR1&2−/− mice. TNFα exerts a protective effect on respiratory function in isolated mitochondria subjected to an anoxia–reoxygenation insult. This effect appears to be independent of its cell surface receptors, but is likely to be mediated by ROS and sphingolipids.

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Acknowledgments

This work was supported in part by a CRIG grant from the Wellcome, the Interuniversity Cape Heart Group of the South African Medical Research Council and the National Research Foundation. S.L. was supported by a Servier Senior Fellowship for Research in Heart Failure and a Medical Research Council career award. L.L. by the Wellcome Trust and the Medical Research Council of South Africa.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Cardioprotection Group, Hatter Cardiovascular Research Institute, Department of Medicine, Faculty of Health Sciences, University of Cape Town, Anzio Road, Observatory 7925, Cape Town, South Africa

    Lydia Lacerda, Joy McCarthy, Shazia F. K. Mungly, Lionel H. Opie & Sandrine Lecour

  2. Translational Medicine Branch, NHLBI, National Institute of Health, Bethesda, MD, USA

    Edward G. Lynn & Michael N. Sack

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  1. Lydia Lacerda
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Correspondence to Lydia Lacerda.

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Lacerda, L., McCarthy, J., Mungly, S.F.K. et al. TNFα protects cardiac mitochondria independently of its cell surface receptors. Basic Res Cardiol 105, 751–762 (2010). https://doi.org/10.1007/s00395-010-0113-4

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