Bioactivation and Toxicity of Acetaminophen in a Rat Hepatocyte Micropatterned Coculture System

• Published in: Journal of biochemical and molecular toxicology Vol. 27; no. 10; pp. 471 - 478
• Main Authors: Ukairo, Okechukwu, McVay, Michael, Krzyzewski, Stacy, Aoyama, Simon, Rose, Kelly, Andersen, Melvin E., Khetani, Salman R., LeCluyse, Edward L.
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01-10-2013; Wiley Subscription Services, Inc
• Subjects: Acetaminophen; Acetaminophen - toxicity; Adenosine Triphosphate - antagonists & inhibitors; Adenosine Triphosphate - biosynthesis; Albumins - metabolism; Analgesics, Non-Narcotic - toxicity; Animals; Bioactivation; Buthionine Sulfoximine - pharmacology; Coculture Techniques; Cocultures; Cytochrome P-450 CYP3A - metabolism; Dexamethasone - pharmacology; Dose-Response Relationship, Drug; Fibroblasts - cytology; Glutathione - antagonists & inhibitors; Glutathione - metabolism; Hepatocytes - cytology; Hepatocytes - drug effects; Hepatocytes - metabolism; Male; Micropatterning; Models, Biological; Primary Cell Culture; Rat Hepatocytes; Rats; Rats, Sprague-Dawley; Up-Regulation - drug effects; Urea - metabolism; Micropatterning; Rat Hepatocytes; Acetaminophen; Bioactivation; Cocultures
• ISSN: 1095-6670; 1099-0461
• DOI: 10.1002/jbt.21512

ABSTRACT
We have recently shown that primary rat hepatocytes organized in micropatterned cocultures with murine embryonic fibroblasts (HepatoPac™) maintain high levels of liver functions for at least 4 weeks. In this study, rat HepatoPac was assessed for its utility to study chemical bioactivation and associated hepatocellular toxicity. Treatment of HepatoPac cultures with acetaminophen (APAP) over a range of concentrations (0–15 mM) was initiated at 1, 2, 3, or 4 weeks followed by the assessment of morphological and functional endpoints. Consistent and reproducible concentration‐dependent effects on hepatocyte structure, viability, and basic functions were observed over the 4‐week period, and were exacerbated by depleting glutathione using buthionine sulfoximine or inducing CYP3A using dexamethasone, presumably due to increased reactive metabolite‐induced stress and adduct formation. In conclusion, the results from this study demonstrate that rat HepatoPac represents a structurally and functionally stable hepatic model system to assess the long‐term effects of bioactivated compounds. © 2013 Wiley Periodicals, Inc. J BiochemMol Toxicol 27:471‐478, 2013; View this article online at wileyonlinelibrary.com. DOI 10.1002/jbt.21512