Lycopene entrapped in human albumin protects 2'-deoxyguanosine against singlet oxygen damage

Abstract

The generation of electronically excited molecular oxygen 1O2 has been shown to occur in several biological systems, such as photooxidation of a variety of biological compounds and xenobiotics ("photodynamic action") and also enzymatic reactions. The high reactivity of 1O2 with unsaturated compounds, sulfides and amino groups arises from its electrophilicity and relatively long lifetime. Thus, biological targets for 1O2 having the above functional groups include unsaturated fatty acids, proteins, enzymes and DNA. There is interest in the role of nutrition in the prevention and pathogenesis of cancer. Epidemiological studies in humans have suggested that carotenoids aid in cancer prevention. Lycopene and oxycarotenoids are present at significant levels in cells and plasma. Extensively conjugated biomolecules such as carotenoids act largely on physical quenching of 1O2 and in much lesser extent on chemical reaction. In this study we observed the protective effect of beta-carotene and lycopene entrapped in human albumin (HSA) against the oxidative 1O2 attack of 2'-deoxyguanosine (dGuo). Photosensitization with methylene blue associated with Chelex resine or Polymer-Rose bengal (Sensitox) and thermodecomposition of water-soluble endoperoxide 3,3'-(1,4-naphthylidene)dipropionate were employed to generate 1O2. The detection of 8-oxo-7,8-dihydro-2'-deoxyguanosine(8-oxodGuo) and 4-hydroxy-8-oxo-7,8-dihydro-2'-deoxyguanosine(4-OH-8-oxodGuo) were performed using reversed phase HPLC with UV, electrochemical detection and by electrospray ionization mass spectrometry. Results showed a significant decrease in the amount of 8-oxodGuo in the presence of lycopene. The percentages of 4-OH-8-oxodGuo and 8-oxodGuo measured were 50% and 70% lower than the control, respectively. These data indicate that carotenoids entrapped in albumin can be an efficient quencher of 1O2 and may be of interest in protecting against the deleterious effect of this excited state molecule.