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Acetaminophen‐induced injury in HepaRG cells: a novel human cell line for studies of drug hepatotoxicity
Author(s) -
McGill Mitchell R,
Yan Huimin,
Jaeschke Hartmut
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.759.9
Subject(s) - acetaminophen , glutathione , toxicity , metabolite , pharmacology , lactate dehydrogenase , necrosis , programmed cell death , cell culture , mitochondrion , chemistry , liver injury , cell , drug , biochemistry , biology , medicine , apoptosis , enzyme , genetics , organic chemistry
Acetaminophen (APAP) overdose is the most frequent cause of acute liver failure in the U.S. The mechanism of APAP hepatotoxicity in rodents involves formation of a reactive metabolite, glutathione (GSH) depletion, mitochondrial dysfunction, and oncotic necrosis. To investigate these mechanisms in a human system, a metabolically competent cell line is needed. In this study, we tested the value of a human hepatoma‐derived cell line (HepaRG) for APAP toxicity studies. Cells were treated with 20 mM APAP and the time course of cell dysfunction and injury was evaluated. APAP caused a decline in cellular GSH levels to 31% of control at 6 h and 24 % at 24 h. The mitochondrial membrane potential (JC‐1) was unaffected at 6 h but was reduced to 45% of control at 12 h and 20% at 24 h. Lactate dehydrogenase (LDH) release was not observed until 24 h (38%) and 48 h (69%) indicating cell necrosis. In addition, a clear dose‐response (5–20 mM) was observed with each of these parameters. Inhibition of drug metabolism with 2% DMSO abrogated all effects. Experiments with HepG2 cells, which are not metabolically competent, showed no GSH depletion, mitochondrial dysfunction or cell injury. Conclusion: APAP toxicity in HepaRG cells mimics closely the sequence of events observed in rodents. Thus, HepaRG cells may be a useful human model for mechanistic studies of drug hepatotoxicity. (Supported by NIH grant DK070195 and Biopredic Int.)