title: Cellular target of cyclophosphamide toxicity in the murine liver: role of glutathione and site of metabolic activation.
abstract: Hepatic venoocclusive disease (HVOD) is caused by the disruption of the microcirculation by an as-yet unknown mechanism. Previous in vitro studies with azathioprine, monocrotaline, and dacarbazine suggested that toxins that cause HVOD initially causing HVOD target sinusoidal endothelial cells (SEC) perhaps via profound glutathione (GSH) depletion. The current study examines cyclophosphamide toxicity in SEC and hepatocytes, as well as the interplay between the two cell types. Cyclophosphamide was not directly toxic to SEC, but in coculture of SEC and hepatocytes, cyclophosphamide was significantly more toxic to SEC. Two cyclophosphamide metabolites, 4-hydroperoxycyclophosphamide and acrolein, were equally toxic to SEC, and toxicity occurred at 20-fold-lower concentrations than in hepatocytes. 4-Hydroperoxycyclophosphamide depleted GSH by greater than 95% before inducing cell death in SEC. When hepatocyte-GSH levels were sustained with supplemental methionine and serine in coculture, toxicity in both cell types was diminished. In coculture, SEC are significantly more susceptible than hepatocytes to cyclophosphamide toxicity, and this is likely caused by acrolein generated by the hepatocyte. As seen with other toxins implicated in HVOD, the profound depletion of SEC GSH precedes the onset of toxicity. The degree of cyclophosphamide toxicity induced in SEC is determined by both metabolic activation and GSH detoxification in the hepatocytes.
authors.affiliation: Center for Liver Diseases and the Division of Gastrointestinal and Liver Diseases, Department of Medicine, USC School of Medicine, Los Angeles, CA, USA.
authors.forename: L D
authors.initials: LD
authors.lastname: DeLeve
mesh_terms: D000171:Acrolein / Q000633:toxicity; D000818:Animals; D001711:Biotransformation; D018920:Coculture Techniques; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000378:metabolism / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004727:Endothelium / Q000166:cytology / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism*; D006504:Hepatic Veno-Occlusive Disease / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000378:metabolism / Q000633:toxicity*; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D008715:Methionine / Q000494:pharmacology; D051379:Mice; D008809:Mice, Inbred C3H; D012694:Serine / Q000494:pharmacology; D013997:Time Factors
chemical_list: D007166:Immunosuppressive Agents; C012358:4-hydroxycyclophosphamide; D012694:Serine; D000171:Acrolein; D003520:Cyclophosphamide; D008715:Methionine; D005978:Glutathione