ToxSci Advance Access originally published online on May 4, 2009
Toxicological Sciences 2009 110(1):84-94; doi:10.1093/toxsci/kfp090
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Ochratoxin A–Mediated DNA and Protein Damage: Roles of Nitrosative and Oxidative Stresses
* Quality and Safety Department, Nestlé Research Center, Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland
Laboratoire d'Etude du Métabolisme des Médicaments, DSV/iBiTec-S/SPI, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France
1 To whom correspondence should be addressed at Nestle Research Center, PO Box 44, Vers-chez-les Blanc, CH-1000 Lausanne 26, Switzerland. Fax: +41-21-785-85-53. E-mail: christophe.cavin{at}rdls.nestle.com.
Received January 8, 2009; accepted April 27, 2009
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Abstract |
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Ochratoxin A (OTA) is a mycotoxin occurring in a variety of foods. OTA is nephrotoxic and nephrocarcinogenic in rodents. An OTA-mediated increase of the inducible nitric oxide synthase (iNOS) expression was observed in normal rat kidney renal cell line and in rat hepatocyte cultures, suggesting the induction of nitrosative stress. This was associated with an increased nuclear factor kappa-light chain enhancer of activated B cells activity. The potential consequences of iNOS induction were further investigated. A significant increase in the levels of protein nitrotyrosine residues was observed with OTA. In addition, OTA was found to increase the level of DNA abasic sites in both cell cultures system. This end point was used as an indirect measure of 8-nitroguanine formation. Treatment of the cells with L-N6-(1-iminoethyl) lysine, a specific inhibitor of iNOS activity, inhibited the OTA-mediated overnitration of proteins but did not reduce the level of DNA abasic sites. It was found previously that nuclear factor-erythroid 2 p45-related factor 2 (Nrf2) activators were able to restore the cellular defense against oxidative stress and could prevent DNA abasic sites in cell cultures. In the present study, pretreatment of the cells with activators of Nrf2 prevented OTA-mediated increase in lipid peroxidation, confirming the potential of Nrf2 activators to confer protection against OTA-mediated oxidative stress. In addition, it was found that Nrf2 activators could also prevent OTA-induced protein nitration and cytotoxicity. In conclusion, the present data further confirm oxidative stress as a key source of OTA-induced DNA damage and provide additional evidence for a role of this mechanism in OTA carcinogenicity. The exact role of nitrosative stress still remains to be established.
Key Words: mycotoxin; ochratoxin A; inducible nitric oxide synthase; nuclear factor-erythroid 2 p45-related factor 2; oxidative and nitrosative stress; nephrotoxicity; carcinogenicity.
2 These authors contributed equally to this study.