Cobalt-mediated oxidative DNA damage and its prevention by polyphenol antioxidants

Abstract

Although cobalt is a required nutrient, it is toxic due to its ability to generate reactive oxygen species (ROS) and damage DNA. ROS generation by Co²⁺ often has been compared to that of Fe²⁺ or Cu⁺, disregarding the reduction potential differences among these metal ions. In plasmid DNA damage studies, a maximum of 15% DNA damage is observed with Co²⁺/H₂O₂ treatment (up to 50 μM and 400 μM, respectively) significantly lower than the 90% damage observed for Fe²⁺/H₂O₂ or Cu⁺/H₂O₂ treatment. However, when ascorbate is added to the Co²⁺/H₂O₂ system, a synergistic effect results in 90% DNA damage. DNA damage by Fe²⁺/H₂O₂ can be prevented by polyphenol antioxidants, but polyphenols both prevent and promote DNA damage by Cu⁺/H₂O₂. When tested for cobalt-mediated DNA damage affects, eight of ten polyphenols (epicatechin gallate, epigallocatechin gallate, propyl gallate, gallic acid, methyl-3,4,5-trihydroxybenzoate, methyl-4,5-dihydroxybenzoate, protocatechuic acid, and epicatechin) prevent cobalt-mediated DNA damage with IC₅₀ values of 1.3 to 27 μM and two (epigallocatechin and vanillic acid) prevent little to no DNA damage. EPR studies demonstrate cobalt-mediated formation of ^•OH, O₂^•-, and ^•OOH, but not ¹O₂ in the presence of H₂O₂ and ascorbate. Epigallocatechin gallate and methyl-4,5-dihydroxybenzoate significantly reduce ROS generated by Co²⁺/H₂O₂/ascorbate, consistent with their prevention of cobalt-mediated DNA damage. Thus, while cobalt, iron, and copper are all d-block metal ions, cobalt ROS generation and its prevention is significantly different from that of iron and copper.

Keywords: Ascorbate; Cobalt; DNA damage; Hydrogen peroxide; Polyphenol; Reactive oxygen species.