title: Reduced cytochrome P4501A activity and recovery from oxidative stress during subchronic benzo[a]pyrene and benzo[e]pyrene treatment of rainbow trout.
abstract: This study assessed the role of aryl hydrocarbon receptor (AHR) affinity, and cytochrome P4501A (CYP1A) protein and activity in polyaromatic hydrocarbon (PAH)-induced oxidative stress. In the 1-100nM concentration range benzo[a]pyrene (BaP) but not benzo[e]pyrene (BeP) competitively displaced 2nM [(3)H]2, 3, 7, 8-tetrachloro-dibenzo-p-dioxin from rainbow trout AHR2α. Based on appearance of fluorescent aromatic compounds in bile over 3, 7, 14, 28 or 50days of feeding 3μg of BaP or BeP/g fish/day, rainbow trout liver readily excreted these polyaromatic hydrocarbons (PAHs) and their metabolites at near steady state rates. CYP1A proteins catalyzed more than 98% of ethoxyresorufin-O-deethylase (EROD) activity in rainbow trout hepatic microsomes. EROD activity of hepatic microsomes initially increased and then decreased to control activities after 50days of feeding both PAHs. Immunohistochemistry of liver confirmed CYP1A protein increased in fish fed both PAHs after 3days and remained elevated for up to 28days. Neither BaP nor BeP increased hepatic DNA adduct concentrations at any time up to 50days of feeding these PAHs. Comet assays of blood cells demonstrated marked DNA damage after 14days of feeding both PAHs that was not significant after 50days. There was a strong positive correlation between hepatic EROD activity and DNA damage in blood cells over time for both PAHs. Neither CYP1A protein nor 3-nitrotyrosine (a biomarker for oxidative stress) immunostaining in trunk kidney were significantly altered by BaP or BeP after 3, 7, 14, or 28days. There was no clear association between AHR2α affinity and BaP and BeP-induced oxidative stress.
authors.affiliation: Oregon State University, Department of Environmental and Molecular Toxicology, Corvallis, OR, USA. larry.curtis@oregonstate.edu
authors.forename: Lawrence R
authors.initials: LR
authors.lastname: Curtis
authors.forename: Claudia B
authors.initials: CB
authors.lastname: Garzon
authors.forename: Mary
authors.initials: M
authors.lastname: Arkoosh
authors.forename: Tracy
authors.initials: T
authors.lastname: Collier
authors.forename: Mark S
authors.initials: MS
authors.lastname: Myers
authors.forename: Jon
authors.initials: J
authors.lastname: Buzitis
authors.forename: Mark E
authors.initials: ME
authors.lastname: Hahn
mesh_terms: D000818:Animals; D001564:Benzo(a)pyrene / Q000008:administration & dosage / Q000494:pharmacology*; D001580:Benzopyrenes / Q000008:administration & dosage / Q000494:pharmacology*; D019363:Cytochrome P-450 CYP1A1 / Q000187:drug effects / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*
chemical_list: D001580:Benzopyrenes; D001564:Benzo(a)pyrene; C026487:benzo(e)pyrene; D019363:Cytochrome P-450 CYP1A1