High concentrations of genistein exhibit pro-oxidant effects in primary muscle cells through mechanisms involving 5-lipoxygenase-mediated production of reactive oxygen species

Genistein, a typical soy isoflavone, is an important antioxidant for improving human health and animal production but the compound possesses some pro-oxidant potential. In order to explore the latter, the dose–response relationship of various concentrations of genistein on both cellular proliferation and the redox system were examined. The proliferation of primary muscle cells was promoted by a low concentration of genistein but was inhibited by high concentrations, which also enhanced lipid oxidation and suppressed membrane fluidity. By selecting a high concentration (200 μM) as a pro-oxidant treatment, the mechanism underlying the pro-oxidant function of genistein was then explored. The generation of intracellular reactive oxygen species (ROS) was stimulated by 200 μM genistein, with inhibited expression of NADPH oxidase 4 and cyclooxygenase 1 and 2 as well as increased activity of the glutathione redox system. The cellular expression of 5-lipoxygenase, however, was up-regulated by 200 μM genistein and the addition of 5-lipoxygenase inhibitor (Zileuton) decreased genistein-induced intracellular ROS level, close to that from the addition of the ROS scavenger, N-acetylcysteine. It is concluded that higher concentrations of genistein exert pro-oxidant potential in the primary muscle cells through enhancing ROS production in a 5-lipoxygenase-dependent manner.