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

Rola Barhoumi, Youssef Mouneimne, Ernesto Ramos, Christophe Morisseau, Bruce D. Hammock, Stephen Safe, Alan R. Parrish, Robert C. Burghardt

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)45-56
Number of pages12
JournalToxicology and Applied Pharmacology
Volume253
Issue number1
DOIs
StatePublished - May 15 2011

Fingerprint

Benzo(a)pyrene
Polycyclic Aromatic Hydrocarbons
Metabolites
Metabolism
Liver
Spectrum analysis
Rats
Cells
Cell Line
Photons
Epoxide Hydrolases
Optical Imaging
Epoxy Compounds
Carcinogens
Microscopy
Homeostasis
Imaging techniques
Wounds and Injuries
Dynamic analysis
Microscopic examination

Keywords

  • Aryl hydrocarbon receptor
  • BaP metabolites
  • Benzo[a]pyrene
  • EROD activity
  • Microsomal epoxide hydrolase inhibitor
  • Multiphoton spectral analysis

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

Barhoumi, R., Mouneimne, Y., Ramos, E., Morisseau, C., Hammock, B. D., Safe, S., ... Burghardt, R. C. (2011). Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line. Toxicology and Applied Pharmacology, 253(1), 45-56. https://doi.org/10.1016/j.taap.2011.03.009

Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line. / Barhoumi, Rola; Mouneimne, Youssef; Ramos, Ernesto; Morisseau, Christophe; Hammock, Bruce D.; Safe, Stephen; Parrish, Alan R.; Burghardt, Robert C.

In: Toxicology and Applied Pharmacology, Vol. 253, No. 1, 15.05.2011, p. 45-56.

Research output: Contribution to journalArticle

Barhoumi, R, Mouneimne, Y, Ramos, E, Morisseau, C, Hammock, BD, Safe, S, Parrish, AR & Burghardt, RC 2011, 'Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line', Toxicology and Applied Pharmacology, vol. 253, no. 1, pp. 45-56. https://doi.org/10.1016/j.taap.2011.03.009
Barhoumi, Rola ; Mouneimne, Youssef ; Ramos, Ernesto ; Morisseau, Christophe ; Hammock, Bruce D. ; Safe, Stephen ; Parrish, Alan R. ; Burghardt, Robert C. / Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line. In: Toxicology and Applied Pharmacology. 2011 ; Vol. 253, No. 1. pp. 45-56.
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