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.