Characterization of Antioxidant Properties of Natural Killer-Enhancing Factor-B and Induction of Its Expression by Hydrogen Peroxide

Abstract

Natural killer-enhancing factor B (NKEF-B) belongs to a highly conserved family of recently discovered antioxidants. The role of NKEF-B as an antioxidant was demonstrated by its protection of transfected cells to oxidative damage by hydrogen peroxide. To further characterize the antioxidant properties of NKEF-B, we compared the sensitivity of a human endothelial cell line ECV304 and its transfectant, B/1 that hyperexpresses NKEF-B, to various oxidants. In addition, we investigated the changes in the expression of NKEF-B mRNA upon oxidative stress. We found that B/1 was significantly more resistant than the control cells to the oxidative stresses caused byt-butyl hydroperoxide (t-BHP) and methyl mercury (MeHg). In contrast, there was no difference in the sensitivity of B/1 and the control cells to sulfhydryl reactive agents, diethyl maleate and diamide. B/1 was also as sensitive as the control cells to buthionine sulfoximine. The expression of NKEF-B mRNA was induced when the parental cell line ECV304 was treated with 2 mmHP. The induction reached a maximum level around 2 hr and decreased to the basal level around 4 hr. NKEF-A mRNA was not induced by HP. These results demonstrate antioxidant activities of NKEF-B toward prooxidants such as alkyl hydroperoxide and MeHg. Together with its antioxidant activity, the induction of NKEF-B by HP indicates that NKEF-B is an important oxidative stress protein providing protection against a variety of xenobiotic toxic agents.