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Measurement of myocardial free radical production during exercise using EPR spectroscopy

¹Cardiovascular Division, University of Minnesota Medical School; ²Minneapolis Heart Institute Foundation at Abbott Northwestern Hospital; and ³Department of Biochemistry, University of Minnesota, Minneapolis, Minnesota

Submitted 25 April 2005 ; accepted in final form 9 January 2006

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 seven chronically instrumented dogs during rest and treadmill exercise (6.4 km/h at 10° grade; and heart rate, 204 ± 3 beats/min) using electron paramagnetic resonance spectroscopy in conjunction with the spin trap -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 min and extracted with toluene. The hyperfine splitting constants of the radicals were _N = 13.7 G and _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 x 10⁸ arbitrary units (P = not significant). 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.

spin trap; coronary blood flow; myocardial oxygen consumption; electron paramagnetic resonance

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