Prooxidant action of rhodizonic acid: transition metal-dependent generation of reactive oxygen species causing the formation of 8-hydroxy-2'-deoxyguanosine formation in DNA.

Keiko Murakami, Miyako Haneda, Makoto Naruse, Masataka Yoshino

Index: Toxicol. In Vitro 20(6) , 910-4, (2006)

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Abstract

Rhodizonic acid, a six-membered cyclic hydroxyquinone, produced reactive oxygen species as a complex with transition metals. Addition of rhodizonic acid with ferrous ion caused an inactivation of aconitase the most sensitive enzyme to oxidative stress in permeabilized yeast cells. The iron-dependent inactivation of aconitase implies the rhodizonic acid/iron-mediated generation of reactive oxygen species. Spectrophotometric analysis of the interaction of rhodizonic acid with FeSO4 showed that addition of superoxide dismutase could inhibit the oxidation of rhodizonic acid, suggesting that reactive oxygen species produced from rhodizonic acid is superoxide radical. Rhodizonic acid further acted as a prooxidant causing a copper-dependent DNA damage. Treatment of DNA from plasmid pBR322 and calf thymus with rhodizonic acid plus copper caused strand scission and the formation of 8-hydroxy-2'-deoxyguanosine in DNA. Addition of catalase protected DNA from the rhodizonic acid-mediated strand scission, indicating that hydroxyl radical may participate in the DNA damage. Rhodizonic acid also showed a potent copper-reducing activity. These results indicate that copper ion reduced by rhodizonic acid may participate in the formation of superoxide radical that converts to hydrogen peroxide and hydroxyl radical. Other cyclic hydroxyquinones such as four-membered squaric acid and five-membered croconic acid did not show any prooxidant and reducing effects. Cytotoxic effects of tetrahydroquinone the precursor of rhodizonic acid may be related to the prooxidant properties of rhodizonic acid formed in cells.