title: Interactive effects of zearalenone and its metabolites on cytotoxicity and metabolization in ovarian CHO-K1 cells.
abstract: Zearalenone (ZEA) is a non-steroidal estrogen mycotoxin with high binding affinity to estrogen receptors. ZEA is rapidly absorbed and metabolized in vivo to α-zearalenol (α-ZOL) and β-zearalenol (β-ZOL). So, mixtures of them may be present in biological systems and suppose a hazard to animals and human health. The aims of this study were to determine the cytotoxic effects of ZEA and its metabolites, alone and in combination in ovarian (CHO-K1) cells during 24, 48 and 72h by the MTT assay; and to investigate the metabolism of the CHO-K1 cells on ZEA, and its conversion into α-ZOL and β-ZOL by CHO-K1 cell after 24 and 48h of exposure. The IC50 value obtained for individual mycotoxins range from 60.3 to >100.0μM, from 30.0 to 33.0μM and from 55.0 to >75.0μM for ZEA, α-ZOL and β-ZOL, respectively. Cytotoxic interactions were assayed by the isobologram method, which provides a combination index (CI) value as a quantitative measure of the degree of the three mycotoxin interaction. The CI values for binary combinations ranged from 0.56±0.15 (synergism at low concentrations) to 5.25±5.10 (addition at high concentrations) and tertiary combinations from 2.95±0.75 (antagonism at low concentrations) to 0.41±0.23 (synergism at high concentrations). The concentration of ZEA and its metabolites was determined with liquid chromatography coupled to the mass spectrometer detector-linear ion trap (LC-MS-LIT). The percentage of ZEA degradation ranged from 4% (24h) to 81% (48h). In the same conditions, α-ZOL and β-ZOL concentration decreased from 8% to 85%. No conversion of ZEA in α-ZOL and β-ZOL was detected. However, at 24h of exposure other degradation products of ZEA and its derived were detected.
authors.affiliation: Laboratory of Food Chemistry and Toxicology, Faculty of Pharmacy, University of Valencia, Av. Vicent Andrés Estellés, s/n, 46100 Burjassot, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Tatay
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Meca
authors.forename: Guillermina
authors.initials: G
authors.lastname: Font
authors.forename: Maria-Jose
authors.initials: MJ
authors.lastname: Ruiz
mesh_terms: D000818:Animals; D016466:CHO Cells; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D004968:Estrogens, Non-Steroidal / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity*; D005260:Female; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry; D009183:Mycotoxins / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity*; D013997:Time Factors; D015025:Zearalenone / Q000008:administration & dosage / Q000378:metabolism / Q000633:toxicity*; D015029:Zeranol / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000633:toxicity
chemical_list: D004968:Estrogens, Non-Steroidal; D009183:Mycotoxins; C029659:zearalenol; D015025:Zearalenone; D015029:Zeranol
keywords: Cytotoxicity; Interactions; LC–MS–LIT; Metabolites; Zearalenone