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Cysteine protects freshly isolated cardiomyocytes against oxidative stress by stimulating glutathione peroxidase

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Abstract

Cysteine has been implicated in myocardial protection, although this is controversial and constrained by limited knowledge about the effects of cysteine at the cellular level. This study tested the hypothesis that a physiologically relevant dose of l-cysteine could be safely loaded into isolated cardiomyocytes leading to improved protection against oxidative stress. Freshly isolated adult rat ventricular cardiomyocytes were incubated for 2 h at 37°C with (cysteine incubated) or without (control) 0.5 mM cysteine prior to washing and suspension in fresh cysteine-free media. Cysteine incubated cells had higher intracellular cysteine levels compared to controls (9.6 ± 0.78 vs. 6.5 ± 0.65 nmol/mg protein, P < 0.02, n = 6 ± SE). Cell homeostasis indicators were similar in the two groups. Cysteine incubated cells had significantly higher glutathione peroxidase (GPx) activity (1.11 ± 0.23 vs. 0.54 ± 0.1 U/mg protein, P < 0.05, n = 5 ± SE) and significantly greater expression of GPx-1 (5.01 ± 0.48 vs. 3.01 ± 0.25 OD units/mm2, P < 0.05, n = 4 ± SE) compared to controls. Upon exposure to H2O2, cysteine incubated cells generated fewer reactive oxygen species and took longer to show contractile changes and undergo hypercontracture. However, when cells were exposed to H2O2 in the presence of 0.05 mM of the GPx inhibitor mercaptosuccinic acid, this increased the control cells’ susceptibility to H2O2 and completely abolished the cysteine mediated protection. These results suggest a new role for cysteine in myocardial protection involving stimulation of glutathione peroxidase.

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Acknowledgements

This study was funded by Project Grant no. PG/05/030 from the British Heart Foundation, and by NIHR Bristol BRU in Cardiovascular Medicine.

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Authors and Affiliations

  1. School of Science and Technology, University of New England, Armidale, NSW, 2351, Australia

    Nicola King

  2. Department of Clinical Science @ South Bristol, Faculty of Medicine and Dentistry, University of Bristol, Bristol Royal Infirmary, Bristol, BS2 8HW, UK

    Nicola King, Hua Lin & M.-Saadeh Suleiman

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  1. Nicola King
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Correspondence to Nicola King.

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King, N., Lin, H. & Suleiman, MS. Cysteine protects freshly isolated cardiomyocytes against oxidative stress by stimulating glutathione peroxidase. Mol Cell Biochem 343, 125–132 (2010). https://doi.org/10.1007/s11010-010-0506-6

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  • DOI: https://doi.org/10.1007/s11010-010-0506-6

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