8-hydroxyguanine and hydroquinone

We investigated the effect of catechol derivatives, including dopa, dopamine, adrenaline and noradrenaline, on DNA damage and the mechanisms of DNA strand breakage and formation of 8-hydroxyguanine (8HOG). The catechol derivatives caused strand breakage of plasmid DNA in the presence of ADP-Fe(3+). The DNA damage was prevented by catalase, mannitol and dimethylsulfoxide, suggesting hydroxyl radical (HO..)-like species are involved in the strand breakage of DNA. Iron chelators, such as desferrioxamine and bathophenanthroline, and reduced glutathione also inhibited the DNA damage. Deoxyribose, a molecule that is used to detect HO,, was not degraded by dopa in the presence of ADP-Fe(3+). By adding EDTA, however, dopa induced the marked deoxyribose degradation in the presence of ADP-Fe(3+), indicating that EDTA may extract iron from ADP-Fe(3+) to catalyze HO. formation by dopa. Thus, EDTA was a good catalyst for HO.-generation, whereas it did not promote the strand breakage of DNA. However, calf thymus DNA base damage, which was detected as 8-HOG formation, was caused by dopa in the presence of EDTA-Fe(3+), but not in the presence of ADP-Fe(3+). The 8HOG formation was also inhibited by catalase and HO. scavengers, indicating that HO&z.rad; was involved in the base damage. These results suggest that DNA strand breakage is due to ferryl species rather than HO., and that 8HOG formation is due to HO. rather than ferryl species.

Topics: Animals; Catechols; Cattle; Dihydroxyphenylalanine; DNA; DNA Damage; Dopamine; Epinephrine; Guanine; Hydroquinones; Norepinephrine; Oxygen Consumption; Plasmids; Resorcinols

A simple means of measuring of 8-hydroxyguanine (8-OHGua) levels in urine was developed. Rat and human urine samples were purified by means of strong cation exchange chromatography (Amberlite CG-120), followed by cellulose partition chromatography (Whatman CF-11). Thereafter, 8-OHGua was determined by means of high performance liquid chromatography (HPLC) using an electro-chemical detector. The level of 8-OHGua in rat urine increased by a factor of 2 to 4 after an intraperitoneal administration of 2-nitropropane (25 mg/kg), paraquat (11.3 mg/kg), or hydroquinone (11 mg/kg). On the other hand, the urine of smokers and persons exposed to air polluted with car exhaust also contained 1.9 and 3.8 fold more 8-OHGua, respectively, than that of control nonsmokers. These results indicated that the amount of 8-OHGua in urine is useful marker for monitoring the level of in vivo oxidative stress.

Topics: Adult; Air Pollution; Animals; Biomarkers; Chromatography; Free Radicals; Guanine; Humans; Hydroquinones; Male; Nitroparaffins; Oxidative Stress; Paraquat; Propane; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Smoking