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Title: Environmentally persistent free radicals inhibit cytochrome P450 activity in rat liver microsomes

Abstract

Combustion processes generate particulate matter that affects human health. When incineration fuels include components that are highly enriched in aromatic hydrocarbons (especially halogenated varieties) and redox-active metals, ultrafine particulate matter containing air-stable, environmentally persistent free radicals (EPFRs) is generated. The exposure to fine EPFRs (less than 2.5 μm in diameter) has been shown to negatively influence pulmonary and cardiovascular functions in living organisms. The goal of this study was to determine if these EPFRs have a direct effect on cytochrome P450 function. This was accomplished by direct addition of the EPFRs to rat liver microsomal preparations and measurement of several P450 activities using form-selective substrates. The EPFRs used in this study were formed by heating vapors from an organic compound (either monochlorophenol (MCP230) or 1,2-dichlorobenzene (DCB230)) and 5% copper oxide supported on silica (approximately 0.2 μm in diameter) to 230 °C under vacuum. Both types of EPFRs (but not silica, physisorbed silica, or silica impregnated with copper oxide) dramatically inhibited the activities of CYP1A, CYP2B, CYP2E1, CYP2D2 and CYP3A when incubated at concentrations less than 0.1 mg/ml with microsomes and NADPH. Interestingly, at the same concentrations, the EPFRs did not inhibit HO-1 activity or the reduction of cytochrome c bymore » NADPH-cytochrome P450 reductase. CYP2D2-selective metabolism by rat liver microsomes was examined in more detail. The inhibition of CYP2D2-selective metabolism by both DCB230- and MCP230-EPFRs appeared to be largely noncompetitive and was attenuated in the presence of catalase suggesting that reactive oxygen species may be involved in the mechanism of inhibition. - Highlights: • Combustion of organic pollutants generates long-lived particulate radicals (EPFRs). • EPFRs inhibit metabolism by all cytochromes P450 tested in rat liver microsomes. • EPFR-mediated inhibition is related to spin content and is sensitive to catalase. • EPFR inhibition of CYP2D2 is noncompetitive with respect to substrate. • Exposure to EPFRs may impair the ability to eliminate xenobiotics.« less

Authors:
;  [1]; ; ; ;  [2];  [1]
  1. Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112 (United States)
  2. Department of Chemistry, Louisiana State University, Baton Rouge, LA 70803 (United States)
Publication Date:
OSTI Identifier:
22439731
Resource Type:
Journal Article
Journal Name:
Toxicology and Applied Pharmacology
Additional Journal Information:
Journal Volume: 277; Journal Issue: 2; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0041-008X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CATALASE; COMBUSTION; CONCENTRATION RATIO; COPPER OXIDES; CYTOCHROMES; ECOLOGICAL CONCENTRATION; HEATING; HEME; LIVER; METABOLISM; MICROSOMES; NANOPARTICLES; NITROPHENOL; POLLUTANTS; PUBLIC HEALTH; RADICALS; RATS; SUBSTRATES; XENOBIOTICS

Citation Formats

Reed, James R., E-mail: rreed@lsuhsc.edu, The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Cawley, George F., Ardoin, Taylor G., The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Dellinger, Barry, Lomnicki, Slawomir M., Hasan, Farhana, Kiruri, Lucy W., Backes, Wayne L., and The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112. Environmentally persistent free radicals inhibit cytochrome P450 activity in rat liver microsomes. United States: N. p., 2014. Web. doi:10.1016/J.TAAP.2014.03.021.
Reed, James R., E-mail: rreed@lsuhsc.edu, The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Cawley, George F., Ardoin, Taylor G., The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Dellinger, Barry, Lomnicki, Slawomir M., Hasan, Farhana, Kiruri, Lucy W., Backes, Wayne L., & The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112. Environmentally persistent free radicals inhibit cytochrome P450 activity in rat liver microsomes. United States. https://doi.org/10.1016/J.TAAP.2014.03.021
Reed, James R., E-mail: rreed@lsuhsc.edu, The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Cawley, George F., Ardoin, Taylor G., The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112, Dellinger, Barry, Lomnicki, Slawomir M., Hasan, Farhana, Kiruri, Lucy W., Backes, Wayne L., and The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112. 2014. "Environmentally persistent free radicals inhibit cytochrome P450 activity in rat liver microsomes". United States. https://doi.org/10.1016/J.TAAP.2014.03.021.
@article{osti_22439731,
title = {Environmentally persistent free radicals inhibit cytochrome P450 activity in rat liver microsomes},
author = {Reed, James R., E-mail: rreed@lsuhsc.edu and The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112 and Cawley, George F. and Ardoin, Taylor G. and The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112 and Dellinger, Barry and Lomnicki, Slawomir M. and Hasan, Farhana and Kiruri, Lucy W. and Backes, Wayne L. and The Stanley S. Scott Cancer Center, Louisiana State University Health Science Center, 533 Bolivar St., New Orleans, LA 70112},
abstractNote = {Combustion processes generate particulate matter that affects human health. When incineration fuels include components that are highly enriched in aromatic hydrocarbons (especially halogenated varieties) and redox-active metals, ultrafine particulate matter containing air-stable, environmentally persistent free radicals (EPFRs) is generated. The exposure to fine EPFRs (less than 2.5 μm in diameter) has been shown to negatively influence pulmonary and cardiovascular functions in living organisms. The goal of this study was to determine if these EPFRs have a direct effect on cytochrome P450 function. This was accomplished by direct addition of the EPFRs to rat liver microsomal preparations and measurement of several P450 activities using form-selective substrates. The EPFRs used in this study were formed by heating vapors from an organic compound (either monochlorophenol (MCP230) or 1,2-dichlorobenzene (DCB230)) and 5% copper oxide supported on silica (approximately 0.2 μm in diameter) to 230 °C under vacuum. Both types of EPFRs (but not silica, physisorbed silica, or silica impregnated with copper oxide) dramatically inhibited the activities of CYP1A, CYP2B, CYP2E1, CYP2D2 and CYP3A when incubated at concentrations less than 0.1 mg/ml with microsomes and NADPH. Interestingly, at the same concentrations, the EPFRs did not inhibit HO-1 activity or the reduction of cytochrome c by NADPH-cytochrome P450 reductase. CYP2D2-selective metabolism by rat liver microsomes was examined in more detail. The inhibition of CYP2D2-selective metabolism by both DCB230- and MCP230-EPFRs appeared to be largely noncompetitive and was attenuated in the presence of catalase suggesting that reactive oxygen species may be involved in the mechanism of inhibition. - Highlights: • Combustion of organic pollutants generates long-lived particulate radicals (EPFRs). • EPFRs inhibit metabolism by all cytochromes P450 tested in rat liver microsomes. • EPFR-mediated inhibition is related to spin content and is sensitive to catalase. • EPFR inhibition of CYP2D2 is noncompetitive with respect to substrate. • Exposure to EPFRs may impair the ability to eliminate xenobiotics.},
doi = {10.1016/J.TAAP.2014.03.021},
url = {https://www.osti.gov/biblio/22439731}, journal = {Toxicology and Applied Pharmacology},
issn = {0041-008X},
number = 2,
volume = 277,
place = {United States},
year = {Sun Jun 01 00:00:00 EDT 2014},
month = {Sun Jun 01 00:00:00 EDT 2014}
}