Abstract
The unique anti-oxidative activity of nitroxide radicals protecting against reactive oxygen-derived species (ROS) has been recently demonstrated in several model systems. The present study focuses on the activity of nitroxide and of its reduced form in cultured rat ventricular cardiomyocytes exposed to O2.- and H2O2 generated by hypoxanthine (HX) and xanthine oxidase (XO). To evaluate cell injury, spontaneous beating, leakage of lactate dehydrogenase (LDH), and depletion of cellular ATP were determined. The protective effect of 4-OH-2,2,6,6-tetramethyl-piperidine-N-oxyl (TPL) was compared with that of 4-OH-2,2,6,6-tetramethyl-1-hydroxypiperidine (TPL-H) and of several common anti-oxidants. A rapid exchange between TPL and TPL-H, is mediated by cellular metabolism and through reactions with ROS. In particular, TPL under O2.- flux is oxidized to oxo-ammonium cation (TPL+) which comproportionates with TPL-H yielding two nitroxide radicals. Because this exchange limits the distinction between the biological activities of TPL and TPL-H, NADH which can reduce TPL+ was included in order to maintain the nitroxide in its reduced form. The results demonstrate that both TPL and TPL-H protect cardiomyocytes against beating loss and LDH lea...Continue Reading
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