Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line

Journal Article · Sun May 15 00:00:00 EDT 2011 · Toxicology and Applied Pharmacology

DOI:https://doi.org/10.1016/j.taap.2011.03.009· OSTI ID:21535297

Barhoumi, Rola ^[1]; Mouneimne, Youssef ^[2]; Ramos, Ernesto ^[1]; Morisseau, Christophe; Hammock, Bruce D ^[3]; Safe, Stephen ^[4]; Parrish, Alan R ^[5]

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1. Department of Veterinary Integrative Biosciences, Texas A and M University, College Station, TX 77843-4458 (United States)
2. American University of Beirut, Beirut (Lebanon)
3. Department of Entomology and Cancer Center, University of California at Davis, Davis, CA 95616 (United States)
4. Department of Veterinary Physiology and Pharmacology, Texas A and M University, College Station, TX 77843 (United States)
5. Department of Pharmacology and Physiology, University of Missouri, Colombia, MO 65211 (United States)

Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an 'advanced unmixing process', identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 h. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to the analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.

• OSTI ID: 21535297
• Journal Information: Toxicology and Applied Pharmacology, Vol. 253, Issue 1; Other Information: DOI: 10.1016/j.taap.2011.03.009; PII: S0041-008X(11)00095-0; Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; ISSN 0041-008X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
CARCINOGENS
EPOXIDES
FLUORESCENCE
HOMEOSTASIS
INJURIES
LIVER CELLS
METABOLISM
METABOLITES
POLYCYCLIC AROMATIC HYDROCARBONS
PYRENE
RATS
RECEPTORS
UPTAKE
ANIMAL CELLS
ANIMALS
AROMATICS
CONDENSED AROMATICS
DISEASES
EMISSION
HYDROCARBONS
LUMINESCENCE
MAMMALS
MEMBRANE PROTEINS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PHOTON EMISSION
PROTEINS
RODENTS
SOMATIC CELLS
VERTEBRATES