title: Cysteine protects freshly isolated cardiomyocytes against oxidative stress by stimulating glutathione peroxidase.
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/mm(2), P < 0.05, n = 4 ± SE) compared to controls. Upon exposure to H(2)O(2), cysteine incubated cells generated fewer reactive oxygen species and took longer to show contractile changes and undergo hypercontracture. However, when cells were exposed to H(2)O(2) in the presence of 0.05 mM of the GPx inhibitor mercaptosuccinic acid, this increased the control cells' susceptibility to H(2)O(2) and completely abolished the cysteine mediated protection. These results suggest a new role for cysteine in myocardial protection involving stimulation of glutathione peroxidase.
authors.affiliation: School of Science and Technology, University of New England, Armidale, NSW, Australia. Nking20@une.edu.au
authors.forename: Nicola
authors.initials: N
authors.lastname: King
authors.forename: Hua
authors.initials: H
authors.lastname: Lin
authors.forename: M-Saadeh
authors.initials: MS
authors.lastname: Suleiman
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000502:physiology*; D004789:Enzyme Activation; D005979:Glutathione Peroxidase / Q000378:metabolism*; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D018384:Oxidative Stress; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D005979:Glutathione Peroxidase; D003545:Cysteine