title: Ochratoxin A-induced cytotoxicity, genotoxicity and reactive oxygen species in kidney cells: An integrative approach of complementary endpoints.
abstract: Ochratoxin A (OTA) is a well-known nephrotoxic and potential carcinogenic agent but no consensus about the molecular mechanisms underlying its deleterious effects has been reached yet. The aim of this study is to integrate several endpoints concerning OTA-induced toxicological effects in Vero kidney cells in order to obtain additional mechanistic data, especially regarding the influence of reactive oxygen species (ROS). One innovative aspect of this work is the use of the superoxide dismutase mimic (SODm) MnTnHex-2-PyP as a mechanistic tool to clarify the involvement of oxidative stress in OTA toxicity. The results showed concentration and time-dependent cytotoxic effects of OTA (crystal violet, neutral red and LDH leakage assays). While the SODm mildly increased cell viability, trolox and ascorbic acid had no effect with regards to this endpoint. OTA induced micronuclei formation. Using the FPG modified comet assay, OTA modestly increased the % of DNA in tail, revealing the presence of oxidative DNA lesions. This mycotoxin increased apoptosis, which was attenuated by SODm. In addition, the SODm decreased the ROS accumulation observed in DHE assay. Taken together, our data suggest that ROS partially contribute to the cytotoxicity and genotoxicity of OTA, although other mechanisms may be relevant in OTA-induced deleterious effects.
authors.affiliation: CBIOS, Universidade Lusófona Research Center for Biosciences & Health Technologies, Campo Grande 376, 1749-024 Lisboa, Portugal; Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisboa, Portugal; Department of Biomedical Sciences, University of Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Alcalá de Henares, Madrid, Spain.
authors.forename: João G
authors.initials: JG
authors.lastname: Costa
authors.affiliation: CBIOS, Universidade Lusófona Research Center for Biosciences & Health Technologies, Campo Grande 376, 1749-024 Lisboa, Portugal.
authors.forename: Nuno
authors.initials: N
authors.lastname: Saraiva
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Patrícia S
authors.initials: PS
authors.lastname: Guerreiro
authors.affiliation: Department of Human Genetics, National Institute of Health Dr. Ricardo Jorge, I.P., Lisboa, Portugal; Center for Toxicogenomics and Human Health (ToxOmics), NMS, FCM - UNL, Lisboa, Portugal.
authors.forename: Henriqueta
authors.initials: H
authors.lastname: Louro
authors.affiliation: Department of Human Genetics, National Institute of Health Dr. Ricardo Jorge, I.P., Lisboa, Portugal; Center for Toxicogenomics and Human Health (ToxOmics), NMS, FCM - UNL, Lisboa, Portugal.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Silva
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Joana P
authors.initials: JP
authors.lastname: Miranda
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Matilde
authors.initials: M
authors.lastname: Castro
authors.affiliation: Department of Radiation Oncology, Duke University Medical School, Durham, NC, USA.
authors.forename: Ines
authors.initials: I
authors.lastname: Batinic-Haberle
authors.affiliation: CBIOS, Universidade Lusófona Research Center for Biosciences & Health Technologies, Campo Grande 376, 1749-024 Lisboa, Portugal. Electronic address: ana.fernandes@ulusofona.pt.
authors.forename: Ana S
authors.initials: AS
authors.lastname: Fernandes
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Professor Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Nuno G
authors.initials: NG
authors.lastname: Oliveira
mesh_terms: D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle; D002470:Cell Survival; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D007668:Kidney / Q000166:cytology* / Q000187:drug effects*; D015394:Molecular Structure; D009152:Mutagenicity Tests; D009793:Ochratoxins / Q000008:administration & dosage / Q000737:chemistry / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D014709:Vero Cells
chemical_list: D009793:Ochratoxins; D017382:Reactive Oxygen Species; C025589:ochratoxin A
keywords: Antioxidants; Cytotoxicity; Genotoxicity; MnTnHex-2-PyP; Ochratoxin A; Reactive oxygen species