title: Evidence for intracellular superoxide formation following the exposure of guinea pig enterocytes to bleomycin.
abstract: Spin trapping of free radicals during the exposure of guinea pig enterocytes to bleomycin (BLM) was investigated using an in vitro cell suspension. The spin traps employed in this study were 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) and 3,3-diethyl-5,5-dimethyl-1-pyrroline-1-oxide (DEDMPO). The hydroxyl radical spin-trapped adduct 2-hydroxy-5,5-dimethyl-1-pyrrolidinyloxyl (DMPO-OH) was observed with DMPO. In the presence of dimethyl sulfoxide (DMSO), the only 2,2,5-trimethyl-1-pyrrolidinyloxyl (DMPO-CH3) observed was that expected from hydroxyl radical formation by the decomposition of the superoxide spin-trapped adduct 2-hydroperoxy-5,5-dimethyl-1-pyrrolidinyloxyl (DMPO-OOH). Production of hydroxyl radical was not detected in the presence of DEDMPO, which is a nitrone that will spin trap hydroxyl radical, but not superoxide, at cellular concentrations. Thus, these data indicate that superoxide was produced during the exposure of guinea pig enterocytes to BLM and that DMPO-OH resulted from the cellular bioreduction of DMPO-OOH by glutathione peroxidase. Addition of superoxide dismutase to the in vitro reaction mixture indicated that superoxide production was intracellular.
authors: Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109.
authors: M J
authors: MJ
authors: Turner
authors: C H
authors: CH
authors: Bozarth
authors: K E
authors: KE
authors: Strauss
mesh_terms: D000818:Animals; D001761:Bleomycin / Q000494:pharmacology*; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D006168:Guinea Pigs; D006878:Hydroxides; D017665:Hydroxyl Radical; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000187:drug effects; D008297:Male; D013481:Superoxides / Q000378:metabolism*
chemical_list: D003497:Cyclic N-Oxides; D006878:Hydroxides; D001761:Bleomycin; D013481:Superoxides; C054821:3,3-diethyl-5,5-dimethylpyrroline 1-oxide; D017665:Hydroxyl Radical; C017245:5,5-dimethyl-1-pyrroline-1-oxide