Received for publication September 21, 2004.
Revised November 16, 2004.
Accepted for publication November 30, 2004.

The CYP2E1 humanized transgenic mouse: role in acetaminophen hepatotoxicity

Abstract

The cytochrome P450 2E1 (CYP2E1) enzyme metabolizes and activates a wide array of toxicological substrates, including alcohols, the widely used analgesic acetaminophen, acetone, benzene, halothane and carcinogens such as azoxymethane and dimethylhydrazine. Most studies on the biochemical and pharmacological actions of CYP2E1 are derived from studies with rodents, rabbits and cultured hepatocytes and therefore extrapolation of the results to humans can be difficult. Creating "humanized" mice by introducing the human CYP2E1 gene into Cyp2e1-null mice can circumvent this disadvantage. A transgenic mouse line expressing the human CYP2E1 gene was established. Western blot and high performance liquid chromatography-mass spectrometry (HPLC-MS) analyses revealed human CYP2E1 protein expression and enzymatic activity in the liver of CYP2E1-humanized mice. Treatment of mice with the CYP2E1 inducer acetone demonstrated that human CYP2E1 was inducible in this transgenic model. The response to the CYP2E1 substrate acetaminophen was explored in the CYP2E1-humanized mice. Hepatoxicity, resulting from CYP2E1-mediated activation of acetaminophen, was demonstrated in the livers of CYP2E1-humanized mice by elevated serum alanine aminotransferase levels, increased hepatocyte necrosis and decreased CYP levels. These data establish that in this humanized mouse model, human CYP2E1 is functional and can metabolize and activate different CYP2E1 substrates such as chlorzoxazone, p-nitrophenol, acetaminophen and acetone. CYP2E1-humanized mice will be of great value for delineating the role of human CYP2E1 in ethanol induced oxidative stress and alcoholic liver damage. They will also function as an important in vivo tool for predicting drug metabolism and disposition and drug-drug interactions of chemicals that are substrates for human CYP2E1.

Key words: CYP2E, cytochrome P450, cytochrome P450 catalyzed oxidations, cytochrome P450 regulation, hepatotoxicity, pharmacokinetics, toxicity, transgenic models

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