Interactive effects of zearalenone and its metabolites on cytotoxicity and metabolization in ovarian CHO-K1 cells

• Published in: Toxicology in vitro 2014-02, Vol.28 (1), p.95-103
• Main Authors: Tatay, Elena, Meca, Giuseppe, Font, Guillermina, Ruiz, Maria-Jose
• Format: Article
• Language: English
• Subjects: Animals; CHO Cells; Chromatography, Liquid; Cricetinae; Cricetulus; Cytotoxicity; Estrogens, Non-Steroidal - administration & dosage; Estrogens, Non-Steroidal - metabolism; Estrogens, Non-Steroidal - toxicity; Female; Inhibitory Concentration 50; Interactions; LC–MS–LIT; Mass Spectrometry; Metabolites; Mycotoxins - administration & dosage; Mycotoxins - metabolism; Mycotoxins - toxicity; Time Factors; Zearalenone; Zearalenone - administration & dosage; Zearalenone - metabolism; Zearalenone - toxicity; Zeranol - administration & dosage; Zeranol - analogs & derivatives; Zeranol - toxicity
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2013.06.025

•Zearalenona and its metabolites alter the viability of CHO-K1 cells.•Higher alteration produced their mixtures.•Binary combinations of Zearalenona, α-zearalenol and β-zearalenol showed additive effects at the higher concentrations.•Tertiary combinations shown antagonism at low concentrations and synergistic effect at higher concentrations.•Zearalenone did not produce α-zearalenol and β-zearalenol in CHO-K1 cells. 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.