UPLC-MS/MS analysis of ochratoxin A metabolites produced by Caco-2 and HepG2 cells in a co-culture system
González Arias, Cyndia A.
Rojas-García, A. E.
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Ochatoxin A (OTA) is one of the most important mycotoxins based on its toxicity. The oral route is the main gateway of entry of OTA into the human body, and specialized epithelial cells constitute the first barrier. The present study investigated the in vitro cytotoxic effect of OTA (5, 15 and 45 μM) and production of OTA metabolities in Caco-2 and HepG2 cells using a co-culture Transwell System to mimic the passage through the intestinal epithelium and hepatic metabolism. The results derived from MTS cell viability assays and transepithelial electrical resistance measurements showed that OTA was slightly cytotoxic at the lowest concentration at 3 h, but significant toxicity was observed at all concentrations at 24 h. OTA metabolites generated in this co-culture were ochratoxin B (OTB), OTA methyl ester, OTA ethyl ester and the OTA glutathione conjugate (OTA-GSH). OTA methyl ester was the major metabolite found in both Caco-2 and HepG2 cells after all treatments. Our results showed that OTA can cause cell damage through several mechanisms and that the OTA exposure time is more important that the dosage in in vitro studies. OTA methyl ester is proposed as an OTA exposure biomarker, although future studies should be conducted.
Is part ofFood and Chemical Toxicology, 2017, vol. 109, p. 333-340
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González Arias, Cyndia A.; Marín Sillué, Sònia; Rojas-García, A. E.; Sanchís Almenar, Vicente; Ramos Girona, Antonio J. (Elsevier, 2016)
Modulation of the xenobiotic transformation system and inflammatory response by ochratoxin A exposure using a co-culture system of Caco-2 and HepG2 cells González Arias, Cyndia A.; Crespo Sempere, Ana; Marín Sillué, Sònia; Sanchís Almenar, Vicente; Ramos Girona, Antonio J. (Elsevier, 2015)Cytotoxicity of ochratoxin A (OTA) was evaluated using the MTS assay, and membrane integrity was measured using transepithelial electrical resistance (TEER). A transwell system was used to investigate the effect of OTA on ...
González Arias, Cyndia A.; Piquer Garcia, I.; Marín Sillué, Sònia; Sanchís Almenar, Vicente; Ramos Girona, Antonio J. (Wageningen Academic Publishers, 2014)Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium species with immunosuppressive, teratogenic, and carcinogenic properties. It has been determined that wine is the second largest source of OTA (10% ...