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Biochemical behaviour of norbixin during in vitro DNA damage induced by reactive oxygen species

Published online by Cambridge University Press:  09 March 2007

Karla Kovary ,
Tatiana S. Louvain ,
Maria C. Costa e Silva ,
Franco Albano ,
Barbara B. M. Pires ,
Gustavo A. T. Laranja ,
Celso L. S. Lage  and
Israel Felzenszwalb
Karla Kovary*
Affiliation:
Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Tatiana S. Louvain
Affiliation:
Departamento de Biofísica e Biometria, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Maria C. Costa e Silva
Affiliation:
Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Franco Albano
Affiliation:
Departamento de Biofísica e Biometria, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Barbara B. M. Pires
Affiliation:
Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Gustavo A. T. Laranja
Affiliation:
Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
Celso L. S. Lage
Affiliation:
Instituto de Biofísica, Centro de Ciências da Saúde, Cidade Universitária, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil
Israel Felzenszwalb
Affiliation:
Departamento de Biofísica e Biometria, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brasil
*
*Corresponding author: Karla Kovary, fax +55 21 587-6136, email [email protected]
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Abstract

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Naturally occurring antioxidants such as carotenoids are extensively studied for their potential in reducing the risk for cancer and other chronic diseases. In the present study, the radical-scavenger activity of the food additive norbixin, a water-soluble carotenoid extracted from Bixa orellana seeds and commercialized as annatto, was evaluated under conditions of DNA damage induced by reactive oxygen species, particularly by hydroxyl radicals. The cell-free scavenger activity of norbixin was evaluated using plasmid DNA as target molecule and Sn2+ or Fe2+ as oxidant. The addition of H2O2 enhanced DNA breakage induced by metal ions, particularly Fe2+. Under these conditions, norbixin started to protect plasmid DNA against single- and double-strand breakage at a metal:norbixin ratio of 1:1 (Sn2+) and 1:10 (Fe2+). However, at lower ratios to Sn2+, norbixin enhanced Sn2+-induced DNA breakage (P<0.05). The ability of norbixin to protect genomic DNA against oxidative damage was assessed in murine fibroblasts submitted to H2O2-induced oxidative stress and the results were evaluated by the comet assay. Under low serum conditions (2 % fetal bovine serum (FBS)), a protective effect of norbixin against H2O2-induced DNA breakage was inversely related to its concentration, a protection ranging from 41 % (10 μM) TO 21 % (50 μm). At higher concentrations of norbixin, however, oxidative DNA breakage was still enhanced, even in the presence of a high serum concentration (10 % FBS). Under normal conditions, norbixin per se has no detectable genotoxic or cytotoxic effects on murine fibroblasts. The antimutagenic potential of norbixin against oxidative mutagens was also evaluated by the Salmonella typhimurium assay, with a maximum inhibition of 87 % against the mutagenicity induced by H2O2. Although plasmid DNA and Ames data indicated that norbixin can protect DNA against oxidative damage, it seems to be a risky guardian of genomic DNA as it can also increase the extent of oxidative damage.

Keywords

NorbixinAnnattoSoluble carotenoidAntioxidantPro-oxidant
Type
Research Article
Information
British Journal of Nutrition , Volume 85 , Issue 4 , April 2001 , pp. 431 - 440
Copyright
Copyright © The Nutrition Society 2001

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