Metabolism and lack of DNA reactivity of the mycotoxin ochratoxin a in cultured rat and human primary hepatocytes.
It is still unclear whether the carcinogenic mycotoxin ochratoxin A (OTA) is bioactivated to DNA-binding metabolites in rodents and humans. Therefore, we have incubated cultured rat and human primary hepatocytes with noncytotoxic concentrations of (3)H-OTA ranging from 10(-7) to 10(-5) M for 8 h and determined its metabolism and covalent DNA binding. In rat hepatocytes, OTA was metabolized to small amounts of three products, which were further studied by electrospray ionization (ESI)-MS/MS techniques. In addition to 4-hydroxy-OTA, which is a known product of OTA biotransformation, two novel metabolites were detected and tentatively identified as hexose and pentose conjugates of OTA. The in vitro induction with 3-methylcholanthrene (3MC) increased the formation of 4-hydroxy-OTA but did not alter the formation of the conjugated metabolites. No covalent binding of (3)H-OTA or its metabolites to DNA was observed in rat hepatocytes with or without 3MC induction with a limit of detection of 2 adducts per 10(9) nucleotides. However, the cellular ratio of reduced glutathione to oxidized glutathione was significantly decreased by treatment with OTA. In cultured human hepatocytes, (3)H-OTA was only very poorly metabolized, and no covalent DNA binding was observed. In conclusion, the results of this in vitro study do not support the notion that OTA has the potential to undergo metabolic activation and form covalent DNA adducts in rodents and humans.
Gross-Steinmeyer
Kerstin
K
Drug Metabolism and Pharmacokinetics, Knoll GmbH, P.O. Box 210805, D-67008 Ludwigshafen, Germany.
Weymann
Jürgen
J
Hege
Hans-Günter
HG
Metzler
Manfred
M
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism*; D005978:Glutathione / Q000032:analysis / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D009793:Ochratoxins / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization; D014316:Tritium
D009793:Ochratoxins; D014316:Tritium; C025589:ochratoxin A; D008748:Methylcholanthrene; D004247:DNA; D005978:Glutathione; C025650:4-hydroxyochratoxin A