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Am J Physiol Heart Circ Physiol (January 13, 2006). doi:10.1152/ajpheart.00412.2005
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Submitted on April 25, 2005
Accepted on January 9, 2006

Measurement of Myocardial Free Radical Production During Exercise Using EPR Spectrscopy

Jay H Traverse1*, Yuri E Nesmelov2, Melanie Crampton3, Paul Lindstrom3, David D Thomas2, and Robert J Bache3

1 Medicine, University of Minnesota, Minneapolis, MN, USA; Biochemistry, University of Minnesota, Minneapolis, MN, USA
2 Cardiology, Minneapolis Heart Institute at Abbott Northwestern Hospital, Minneapolis, MN, USA
3 Medicine, University of Minnesota, Minneapolis, MN, USA

* To whom correspondence should be addressed. E-mail: trave004{at}umn.edu.

Exercise is associated with an increase in oxygen flux through the mitochondrial electron transport chain that has recently been demonstrated to increase the production of reactive oxygen species (ROS) in skeletal muscle. This study examined whether exercise also causes free radical production in the heart. We measured ROS production in 7 chronically instrumented dogs during rest and treadmill exercise (6.4 Km/H, 10°grade; heart rate 204 ± 3 beats/min) using EPR spectroscopy in conjunction with the spin trap {alpha}-phenyl - tert -butylnitrone (PBN) (0.14 mol/L) in blood collected from the aorta and coronary sinus (CS). To improve signal detection, the free radical adducts were deoxygenated over a nitrogen stream for 15 minutes and extracted with toluene. The hyperfine splitting constants of the radicals were {alpha}N = 13.7 G and {alpha}H = 1.0 G, consistent with an alkoxyl or carbon centered radical. Resting aortic and CS PBN adduct concentrations were 6.7 and 6.3 (A.U x 108 ) (p=NS). Both aortic and CS adduct concentrations increased during exercise, but there was no significant difference between the aortic and CS concentrations. Thus, in contrast to skeletal muscle, submaximal treadmill exercise did not result in detectable free radical production by the heart.




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