Prooxidant action of rosmarinic acid: Transition metal-dependent generation of reactive oxygen species

Rosmarinic acid and its constituent caffeic acid produced reactive oxygen species in the presence of transition metals. Complex of rosmarinic acid or caffeic acid with iron inactivated aconitase the most sensitive enzyme to oxidative stress. The inactivation of aconitase was iron-dependent, and prevented by TEMPOL, a scavenger of reactive oxygen species, suggesting that the rosmarinic acid/iron-mediated generation of superoxide anion is responsible for the inactivation of aconitase. Direct spectrophotometric determination of hydrogen peroxide and superoxide anion confirmed the rosmarinic acid/iron-dependent production of reactive oxygen species. Treatment of DNA from plasmid pBR322 and calf thymus with rosmarinic acid plus copper caused strand scission and formed 8-hydroxy-2′-deoxyguanosine in DNA. Rosmarinic acid and caffeic acid showed a potent activity that reduces transition metals. These results suggest that transition metals reduced by rosmarinic acid can form superoxide radical by one electron reduction of oxygen molecule: superoxide radical in turn converts to hydrogen peroxide and hydroxyl radical causing the formation of DNA base adduct. Cytotoxicity of rosmarinic acid may be related to the prooxidant action resulting from metal-reducing activity.