title: Degradation of DMPO adducts from hydroxyl and 1-hydroxyethyl radicals by rat liver microsomes.
abstract: Hydroxyl and 1-hydroxyethyl radical adducts of 5,5-dimethylpyrroline N-oxide (DMPO) were prepared by photolysis, and mechanisms for loss of their EPR signals in rat liver microsomal suspensions were evaluated. Rates of NADPH-dependent EPR signal loss were more rapid in phosphate buffer than in Tris buffer. Addition of superoxide dismutase (SOD) partially protected the adducts when Tris was used as a buffer, but was relatively ineffective in the presence of phosphate. The ferrous iron chelator bathophenan-throlene partially protected the spin adducts in the presence and absence of phosphate, but complete protection was observed when SOD was also added. The spin adducts were unstable in the presence of Fe+2 and K3Fe(CN)6, but Fe+3 alone had little effect on the EPR signals. The data are consistent with two mechanisms for microsomal degradation of DMPO spin adducts under these conditions. Microsomes from superoxide in the presence of oxygen and NADPH, which attacks these DMPO spin adducts directly. The spin adducts are also degraded in the presence of Fe+2, and phosphate stimulates this iron-dependent destruction of DMPO spin adducts.
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Oklahoma Health Sciences Center, Oklahoma City 73190, USA. lester-reinke@uokhsc.edu
authors.forename: L A
authors.initials: LA
authors.lastname: Reinke
authors.forename: D R
authors.initials: DR
authors.lastname: Moore
authors.forename: P B
authors.initials: PB
authors.lastname: McCay
mesh_terms: D000818:Animals; D003497:Cyclic N-Oxides / Q000737:chemistry*; D004578:Electron Spin Resonance Spectroscopy; D017665:Hydroxyl Radical / Q000737:chemistry*; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry*; D009249:NADP; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels; D019097:Spin Trapping; D013482:Superoxide Dismutase / Q000737:chemistry; D013481:Superoxides / Q000737:chemistry
chemical_list: D003497:Cyclic N-Oxides; D013113:Spin Labels; D013481:Superoxides; D017665:Hydroxyl Radical; D009249:NADP; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D013482:Superoxide Dismutase