Comparative study of the formation of oxidative damage marker 8-hydroxy-2′-deoxyguanosine (8-OHdG) adduct from the nucleoside 2′-deoxyguanosine by transition metals and suspensions of particulate matter in relation to metal content and redox reactivity

Abstract

An association between exposure to ambient particulate matter (PM) and increased incidence of mortality and morbidity due to lung cancer and cardiovascular diseases has been demonstrated by recent epidemiological studies. Reactive oxygen species (ROS), especially hydroxyl radicals, generated by PM, have been suggested by many studies as an important factor in the oxidative damage of DNA by PM. The purpose of this study was to characterize quantitatively hydroxyl radical generation by various transition metals in the presence of H₂O₂ in aqueous buffer solution (pH 7.4) and hydroxylation of 2′-deoxyguanosine (dG) to 8-hydroxy-2′-deoxyguanosine (8-OHdG) under similar conditions. The order of metals' redox reactivity and hydroxyl radical production was Fe(II), V(IV), Cu(I), Cr(III), Ni(II), Co(II), Pb(II), Cd(II). Then, we investigated the generation of hydroxyl radicals in the presence of H₂O₂ by various airborne PM samples, such as total suspended particulate (TSP), PM₁₀, PM_2.5 (PM with aerodynamic diameter 10 and 2.5 μm), diesel exhaust particles (DEP), gasoline exhaust particles (GEP) and woodsmoke soot under the same conditions. When suspensions of PMs were incubated with H₂O₂ and dG at pH 7.4, all particles induced hydroxylation of dG and formation of 8-OHdG in a dose–dependent increase. Our findings demonstrated that PM's hydroxyl radical (HO√) generating ability and subsequent dG hydroxylation is associated with the concentration of water-soluble metals, especially Fe and V and other redox or ionizable transition metals and not their total metal content, or insoluble metal oxides, via a Fenton-driven reaction of H₂O₂ with metals. Additionally, we observed, by Electron paramagnetic resonance (EPR), that PM suspensions in the presence of H₂O₂ generated radical species with dG, which were spin–trapped by 2-methyl-2-nitroso–propane (MNP).

• Airborne particulate matter
• Fenton reaction
• hydrogen peroxide
• hydroxyl radicals
• 8-hydroxyl-2 ′-deoxyguanosine
• reactive oxygen species

Abbreviations
DMPO	=	5,5-Dimethyl-1-pyrroline–N-oxide
8-OHdG	=	8-hydroxy-2′-deoxyguanosine
MNP	=	2-methyl-2-nitroso–propane
dG	=	2′-deoxyguanosine
TSP	=	total suspended particulates
DEP	=	diesel exhaust particles
GEP	=	gasoline exhaust particles

Abbreviations
DMPO	=	5,5-Dimethyl-1-pyrroline–N-oxide
8-OHdG	=	8-hydroxy-2′-deoxyguanosine
MNP	=	2-methyl-2-nitroso–propane
dG	=	2′-deoxyguanosine
TSP	=	total suspended particulates
DEP	=	diesel exhaust particles
GEP	=	gasoline exhaust particles