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Nitrogen Compounds Prevent H9c2 Myoblast Oxidative Stress-Induced Mitochondrial Dysfunction and Cell Death

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An Erratum to this article was published on 02 March 2010

Abstract

Oxidative stress has been connected to various forms of cardiovascular diseases. Previously, we have been investigating the potential of new nitrogen-containing synthetic compounds using a neuronal cell model and different oxidative stress conditions in order to elucidate their potential to ameliorate neurodegenerative diseases. Here, we intended to extend these initial studies and investigate the protective role of four of those new synthetic compounds (FMA4, FMA7, FMA762 and FMA796) against oxidative damage induced to H9c2 cardiomyoblasts by tert-butylhydroperoxide (t-BHP). The data indicates that FMA762 and FMA796 decrease t-BHP-induced cell death, as measured by both sulforhodamine B assay and nuclear chromatin condensation evaluation, at non-toxic concentrations. In addition, the two mentioned compounds inhibit intracellular signalling mechanisms leading to apoptotic cell death, namely those mediated by mitochondria, which was confirmed by their ability to overcome t-BHP-induced morphological changes in the mitochondrial network, loss of mitochondrial membrane potential, increased expression of the pro-apoptotic proteins p53, Bax and AIF and activation of caspases-3 and -9. Importantly, our results indicate that the compounds’ ROS scavenging ability plays a crucial role in the protection profile, as a significant decrease in t-BHP-induced oxidative stress occurred in their presence. Data obtained indicates that some of the test compounds may clearly prove valuable in a clinical context by diminishing cardiac injury associated to oxidative stress without any toxicity.

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Abbreviations

ROS:

Reactive oxygen species

t-BHP:

tert-butyl hydroperoxide

SRB:

Sulforhodamine B

TMRM:

Tetramethyl rhodamine methyl ester

Δψ :

Membrane potential

CCCP:

Carbonyl cyanide m-chloro phenyl hydrazone

AIF:

Apoptosis-inducing factor

CM-H2DCFDA:

5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate

NBT:

NitroBlue Tetrazolium

fSD:

Intracellular fluorescence standard deviation

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Acknowledgments

JPS is supported by the Portuguese Foundation for Science and Technology (FCT), Grant SFRH/BD/17174/2004. The present work was supported by FCT research Grant PTDC/QUI/64358/2006. We want to thank Prof. Fernanda Proença, from the Department of Chemistry, University of Minho, for kindly supplying the synthetic nitrogen compounds used.

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Authors and Affiliations

  1. CBMA—Molecular and Environmental Biology Centre, Department of Biology, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    João P. Silva & Olga P. Coutinho

  2. Center for Neurosciences and Cellular Biology, University of Coimbra, Coimbra, Portugal

    Vilma A. Sardão & Paulo J. Olveira

Authors
  1. João P. Silva
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  2. Vilma A. Sardão
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  4. Paulo J. Olveira
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Correspondence to João P. Silva.

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An erratum to this article can be found at http://dx.doi.org/10.1007/s12012-010-9067-x

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Silva, J.P., Sardão, V.A., Coutinho, O.P. et al. Nitrogen Compounds Prevent H9c2 Myoblast Oxidative Stress-Induced Mitochondrial Dysfunction and Cell Death. Cardiovasc Toxicol 10, 51–65 (2010). https://doi.org/10.1007/s12012-010-9062-2

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