Reduction of lipid-soluble nitroxides in CHO cells and macrophage tumor cells

There is a need to understand the metabolism of nitroxides because of their usefulness in measurements in living cells of complex phenomena, such as biophysical properties, redox metabolism, and the concentration of oxygen at specific sites. As part of a systematic study of the metabolism of nitroxides in cells, the authors studied Chinese hamster ovary (CHO) cells and mouse macrophage tumor (M5076) cells, using a set of lipophilic nitroxides based on 5 doxyl stearate: the free acid, the methyl ester of the acid, and a phosphatidylcholine with two doxyl stearates esterified to the glycerol. The rates of metabolism of these nitroxides under anoxia differed significantly as a function of both the type of cell and the type of nitroxide. The rates of reduction of the three lipophilic nitroxides depended on their localization. The rates of reduction were first order for all three lipophilic nitroxides, and the only products detected were the respective hydroxylamines. Effects of freeze-thawing and incubation temperature differed in the two cell lines. The authors conclude that the metabolism of nitroxides in different cell lines can be quite different. This may be especially important in understanding studies using nitroxides in living cells, functional organs, and in vivo.