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Journal of Biomedical Optics • Open Access

Effect of ouabain on metabolic oxidative state in living cardiomyocytes evaluated by time-resolved spectroscopy of endogenous NAD(P)H fluorescence
Author(s): Alzbeta Chorvatova; Anton Mateasik; Dusan Chorvat; Fathia Elzwiei

Paper Abstract

Time-resolved spectrometry of endogenous nicotinamide dinucleotide phosphate [NAD(P)H] fluorescence is a useful method to evaluate metabolic oxidative state in living cells. Ouabain is a well-known pharmaceutical drug used in the treatment of cardiovascular disease, the effects of which on myocardial metabolism were recently demonstrated. Mechanisms implicated in these actions are still poorly understood. We investigate the effect of ouabain on the metabolic oxidative state of living cardiac cells identified by time-resolved fluorescence spectroscopy of mitochondrial NAD(P)H. Spectral unmixing is used to resolve individual NAD(P)H fluorescence components. Ouabain decreased the integral intensity of NAD(P)H fluorescence, leading to a reduced component amplitudes ratio corresponding to a change in metabolic state. We also noted that lactate/pyruvate, affecting the cytosolic NADH gradient, increased the effect of ouabain on the component amplitudes ratio. Cell oxidation levels, evaluated as the percentage of oxidized NAD(P)H, decreased exponentially with rising concentrations of the cardiac glycoside. Ouabain also stimulated the mitochondrial NADH production. Our study sheds a new light on the role that ouabain plays in the regulation of metabolic state, and presents perspective on a noninvasive, pharmaceutical approach for testing the effect of drugs on the mitochondrial metabolism by means of time-resolved fluorescence spectroscopy in living cells.

Paper Details

Date Published: 14 May 2012
PDF: 8 pages
J. Biomed. Opt. 17(10) 101505 doi: 10.1117/1.JBO.17.10.101505
Published in: Journal of Biomedical Optics Volume 17, Issue 10
Show Author Affiliations
Alzbeta Chorvatova, International Laser Ctr. (Slovakia)
Anton Mateasik, International Laser Ctr. (Slovakia)
Dusan Chorvat, International Laser Ctr. (Slovakia)
Fathia Elzwiei, Univ. de Montréal (Canada)


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