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Modulation of apoptosis by sulforaphane is associated with PGC-1α stimulation and decreased oxidative stress in cardiac myoblasts

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Abstract

Sulforaphane is a naturally occurring isothiocyanate capable of stimulating cellular antioxidant defenses and inducing phase 2 detoxifying enzymes, which can protect cells against oxidative damage. Oxidative stress and apoptosis are intimately involved in the pathophysiology of cardiac diseases. Although sulforaphane is known for its anticancer benefits, its role in cardiac cells is just emerging. The aim of the present study was to investigate whether sulforaphane can modulate oxidative stress, apoptosis, and correlate with PGC-1α, a transcriptional cofactor involved in energy metabolism. H9c2 cardiac myoblasts were incubated with R-sulforaphane 5 µmol/L for 24 h. Cell viability, ANP gene expression, oxidative stress and apoptosis markers, and protein expression of PGC-1α were studied. In cells treated with sulforaphane, cellular viability increased (12 %) and ANP gene expression decreased (46 %) compared to control cells. Moreover, sulforaphane induced a significant increase in superoxide dismutase (103 %), catalase (101 %), and glutathione S-transferase (72 %) activity, reduced reactive oxygen species levels (15 %) and lipid peroxidation (65 %), as well as stimulated the expression of the cytoprotective enzyme heme oxygenase-1 (4-fold). Sulforaphane also promoted an increase in the expression of the anti-apoptotic protein Bcl-2 (60 %), decreasing the Bax/Bcl-2 ratio. Active Caspase 3\7 and p-JNK/JNK were also reduced by sulforaphane, suggesting a reduction in apoptotic signaling. This was associated with an increased protein expression of PGC-1α (42 %). These results suggest that sulforaphane offers cytoprotection to cardiac cells by activating PGC1-α, reducing oxidative stress, and decreasing apoptosis signaling.

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Acknowledgment

This work was supported by Brazilian Research Agencies: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Fundação de Amparo à Pesquisa do Rio Grande do Sul (FAPERGS), as well as the Northern Ontario School of Medicine. Technical assistance from Tânia Fernandes, Sílvia Guimarães, and Lucas Barbosa is acknowledged. We acknowledge intellectual contribution from Prof. Dr. Maria Flávia Ribeiro, Cláudio Felipe K. Rocha, and Dr. Carmem Sartório. We would also like to thank the Laboratory of Molecular Biology, Endocrinology and Tumor (Labimet, UFRGS).

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Authors and Affiliations

  1. Laboratory of Cardiovascular Physiology, Institute of Basic Health Science (ICBS), Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil

    Rafael O. Fernandes, Jéssica H. P. Bonetto, Alexandre L. de Castro, Ilma Simoni Brum, Alex Sander R. Araujo & Adriane Belló-Klein

  2. Department of Biology, Lakehead University, Thunder Bay, ON, Canada

    Boran Baregzay & Stephanie Puukila

  3. Medical Sciences Division, Northern Ontario School of Medicine, Lakehead University, Thunder Bay, ON, Canada

    Heidi Forsyth & Neelam Khaper

  4. Department of Physiology, Federal University of Pelotas (UFPel), Pelotas, RS, Brazil

    Paulo C. Schenkel

  5. General and Inorganic Chemistry Division, School of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina

    Susana F. Llesuy

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  1. Rafael O. Fernandes
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Correspondence to Neelam Khaper or Adriane Belló-Klein.

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Fernandes, R.O., Bonetto, J.H., Baregzay, B. et al. Modulation of apoptosis by sulforaphane is associated with PGC-1α stimulation and decreased oxidative stress in cardiac myoblasts. Mol Cell Biochem 401, 61–70 (2015). https://doi.org/10.1007/s11010-014-2292-z

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