Oxygen consumption and oxyradical production from microsomal reduction of aqueous extracts of cigarette tar.

Aqueous extracts of cigarette tar (ACT) contain a quinone-hydroquinone-semiquinone system that undergoes autoxidation to produce superoxide and hydrogen peroxide. In this paper we ask whether the production of activated oxygen species from this quinone system can be enhanced via an NAD(P)H-dependent autoxidation in which electrons are donated from electron transport components of the microsomal system to electron acceptors in ACT such as quinones and/or metal chelates. Therefore, the ability of ACT to enhance the oxidation of NADPH, consumption of oxygen, and production of oxyradicals during microsomal electron transport has been investigated. Rates of NADPH oxidation and oxygen consumption are enhanced in a concentration-dependent manner by both aqueous and dimethyl sulfoxide (DMSO) extracts of tar over that observed in the presence of microsomes alone. Over the time course studied, neither NADPH oxidation nor oxygen consumption by tar extracts is observed in the absence of microsomes. Microsomal NADPH oxidation is enhanced by 1,6- and 1,3-benzo[a]pyrene quinones and by iron-EDTA; per unit weight, the enhancement by these compounds is similar to that of tar. NADPH oxidation in the presence of tar and absence of iron-EDTA is inhibited by about 15% by the addition of deferoxamine to the reaction mixture. Chrysene quinone enhances NADPH oxidation to a much greater extent than does either of the benzo[a]pyrene quinones or tar. Inhibition of tar-stimulated microsomal oxygen uptake by superoxide dismutase, stimulation of oxygen uptake by azide (an inhibitor of catalase), and the presence of spin-trappable superoxide anion radical all indicate the reduction of tar components to superoxide and hydrogen peroxide. The similar effects of tar on NADPH oxidation catalyzed either by microsomes or by purified NADPH-cytochrome P450 reductase suggest that cytochrome P450 reductase is an important locus of electron transfer from microsomes to components of cigarette tar extracts. These data suggest that the cytotoxic and mutagenic effects of smoking may be mediated by metabolic activation of quinones and, to a lesser extent, metal chelates in cigarette tar to oxyradicals.