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Cardiovascular Research Advance Access [Accepted Manuscript] published online on September 18, 2007
Cardiovascular Research, doi:10.1093/cvr/cvm011
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Exercise and tachycardia increase NADPH oxidase and ryanodine receptor-2 activity: Possible role in cardioprotection

Gina Sánchez1, Matías Escobar2, Zully Pedrozo2, Pilar Macho1, Raúl Domenech1, Steffen Härtel3, Cecilia Hidalgo2 and Paulina Donoso2,

1 Programas de Fisiopatología, Instituto de Ciencias Biomédicas y Centro FONDAP de Estudios Moleculares de la Célula, Facultad de Medicina, Universidad de Chile
2 Biología Celular y Molecular, Instituto de Ciencias Biomédicas y Centro FONDAP de Estudios Moleculares de la Célula, Facultad de Medicina, Universidad de Chile
3 Anatomía y Biología del Desarrollo, Instituto de Ciencias Biomédicas y Centro FONDAP de Estudios Moleculares de la Célula, Facultad de Medicina, Universidad de Chile

Corresponding author: Paulina Donoso, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Casilla 70005, Santiago 7, Chile. Telephone: (56-2) 978 6602. FAX: (56-2) 777 6916. Email: pdonoso{at}med.uchile.cl

Aim: Our objective was to investigate in cardiac muscle the contribution of NADPH oxidase to a) ryanodine receptor-2 (RyR2) S-glutathionylation and b) the preconditioning effects of exercise and tachycardia on infarct size following coronary artery occlusion.

Methods: We measured NADPH oxidase activity, RyR2 S-glutathionylation and calcium release kinetics in sarcoplasmic reticulum vesicles isolated from dog ventricular muscle after exercise and tachycardia, plus or minus prior administration of the NADPH oxidase inhibitor apocynin. In ventricular muscle sections, we studied the colocalization of NADPH oxidase and RyR2 by confocal microscopy using fluorescent antibodies. We determined the effect of apocynin on the infarct size produced by occlusion of the descendent anterior coronary artery in animals preconditioned by exercise or tachycardia.

Results: Exercise and tachycardia increased NADPH oxidase activity, RyR2 S-glutathionylation and calcium release rates in isolated sarcoplasmic reticulum vesicles. Cardiac muscle sections displayed significant colocalization of NADPH oxidase and RyR2, suggesting direct and specific effects of reactive oxygen species produced by NADPH oxidase on RyR2 activation. The NADPH oxidase inhibitor apocynin prevented the increase in RyR2 S-glutathionylation, reduced calcium release activity and completely prevented the protective effects of exercise and tachycardia on infarct size.

Conclusions: The loss of cardioprotection induced by the NADPH oxidase inhibitor suggests that reactive oxygen species generated by this enzyme are important mediators of the preconditioning response, which presumably involves NADPH oxidase-induced RyR2 S-glutathionylation.

Time for primary review: 30 days


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