eng Science and Education Publishing American Journal of Food and Nutrition 2374-1163 2018-02-09 6 1 9 13 10.12691/ajfn-6-1-2 AJFN2018612 article Robert Settineri sierraprod@aol.com 1 Jie Zhou 2 Jin Ji 3 Rita R. Ellithorpe 4 Steven Rosenblatt 5 Antonio Jimenez 6 Shigeo Ohta 7 Gonzalo Ferreira 8 Garth L. Nicolson 9 Sierra Productions Research, Irvine, CA USA Brunswick Laboratories, Inc., Southborough, MA USA PulchriBio Intl, Boston, MA USA Tustin Longevity Center, Tustin, CA USA Saint John's Health Center, Santa Monica, CA USA Hope4Cancer Institute, Baja California, Mexico Department of Neurology, Juntendo University, Graduate School of Medicine, Tokyo, Japan Department of Biophysicas, Laboratory of Ion Channels, School of Medicine, Universidad de la Republica, Montevideo, Uruguay Department of Molecular Pathology, The Institute for Molecular Medicine, Huntington Beach, CA USA Hydrogenized water is known to have protective effects on cells and tissues, mainly through its antioxidant activities. Here we examined the protective effects of a commercial source of hydrogenized water on cultured human brain cells. Hydrogenized water was able to protect brain cells from oxidative stress and glutamate toxicity. At H2 concentrations above 0.01 mM the glutathione levels increased in cultured brain cells. The level of glutathione rose from approximately 500 to approximately 850 üM at the maximum dose of hydrogenized water with an EC50 of approximately 0.030 mM. Hydrogenized water was also able to enhance the signaling pathway for oxidative stress response mediated by Nrf2 (Nuclear factor erythroid 2 like factor). Treatment of cells with hydrogenized water at concentrations above 0.01 mM H2 induced activation of Nrf2 (EC50 approximately 0.05 mM). Hydrogenized water was also able to protect brain cells against glutamate toxicity. Using a DNA damage response element, (̦H2AX, to monitor the damage of glutamate toxicity we found that concentrations of H2 above 0.01 mM protected cells from glutamate damage with an EC50 of approximately 0.05 mM H2. These in vitro results demonstrated that hydrogenized water can protect brain cells against common types of damage from oxidative stress and glutamate toxicity. http://pubs.sciepub.com/ajfn/6/1/2/ajfn-6-1-2.pdf hydrogenized water bioassays glutathione oxidative stress Nrf2 glutamate toxicity