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Am J Physiol Endocrinol Metab (November 23, 2004). doi:10.1152/ajpendo.00382.2004
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Submitted on August 17, 2004
Accepted on November 15, 2004

Long-Term Calorie Restriction Reduces Proton Leak and Hydrogen Peroxide Production in Liver Mitochondria

Kevork Hagopian1, Mary-Ellen Harper2, Jesmon J. Ram1, Stephen J. Humble1, Richard Weindruch3, and Jon J. Ramsey1*

1 Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA, USA
2 Department of Biochemistry, Microbiology, and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada
3 Department of Medicine, University of Wisconsin Medical School and Veterans Administration Geriatric Research, Education and Clinical Center, Madison, WI, USA

* To whom correspondence should be addressed. E-mail: jjramsey{at}ucdavis.edu.

Calorie restriction (CR), without malnutrition, increases maximal lifespan in diverse species. It has been proposed that reduction in energy expenditure and reactive oxygen species (ROS) production could be a mechanism for life span extension with CR. As a step towards testing this theory, mitochondrial proton leak, H2O2 production and markers of oxidative stress were measured in liver from FBNF1 rats fed control or 40% CR diets for 12 or 18 months. CR was initiated at 6 months of age. Proton leak kinetics curves, generated from simultaneous measures of oxygen consumption and membrane potential, indicated a decrease in proton leak after 18 months of CR while only a trend towards a proton leak decrease was observed after 12 months. Significant shifts in phosphorylation and substrate oxidation curves also occurred with CR; however, these changes occurred in concert with the proton leak changes. Metabolic control analysis indicated no difference in the overall pattern of control of the oxidative phosphorylation system between control and CR animals. At 12 months, no significant differences were observed between groups for H2O2 production or markers of oxidative stress. However, at 18 months, protein carbonyl content was lower in CR animals, as was H2O2 production when mitochondria were respiring on either succinate alone or pyruvate plus malate in the presence of rotenone. These results indicate that long-term CR lowers mitochondrial proton leak and H2O2 production, and this is consistent with the idea that CR may act by decreasing energy expenditure and ROS production.




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