Hydrogenized Water Effects on Protection of Brain Cells from Oxidative Stress and Glutamate Toxicity

Robert Settineri^1, , Jie Zhou², Jin Ji³, Rita R. Ellithorpe⁴, Steven Rosenblatt⁵, Antonio Jimenez⁶, Shigeo Ohta⁷, Gonzalo Ferreira⁸ and Garth L. Nicolson⁹

¹Sierra Productions Research, Irvine, CA USA

²Brunswick Laboratories, Inc., Southborough, MA USA

³PulchriBio Intl, Boston, MA USA

⁴Tustin Longevity Center, Tustin, CA USA

⁵Saint John's Health Center, Santa Monica, CA USA

⁶Hope4Cancer Institute, Baja California, Mexico

⁷Department of Neurology, Juntendo University, Graduate School of Medicine, Tokyo, Japan

⁸Department of Biophysicas, Laboratory of Ion Channels, School of Medicine, Universidad de la Republica, Montevideo, Uruguay

⁹Department of Molecular Pathology, The Institute for Molecular Medicine, Huntington Beach, CA USA

Cite this paper:
Robert Settineri, Jie Zhou, Jin Ji, Rita R. Ellithorpe, Steven Rosenblatt, Antonio Jimenez, Shigeo Ohta, Gonzalo Ferreira and Garth L. Nicolson. Hydrogenized Water Effects on Protection of Brain Cells from Oxidative Stress and Glutamate Toxicity. American Journal of Food and Nutrition. 2018; 6(1):9-13. doi: 10.12691/ajfn-6-1-2

Abstract

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 μM 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, (γH2AX, 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.

Keywords:
hydrogenized water bioassays glutathione oxidative stress Nrf2 glutamate toxicity

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Beal MF. Mitochondria take center stage in aging and neurodegeneration. Ann Neurol. 2005; 58: 495-505.
[2]	Dias V, Junn E, Mouradian MM. The role of oxidative stress in Parkinson’s disease. J Parkinsons Dis. 2013; 3(4): 461-491.
[3]	Schiavone S, Jaquet V, Trabace L, Krause K-H. Severe life stress and oxidative stress in the brain: from animal models to human pathology. Antioxid Redox Signal. 2013; 18(12): 1475-1490.
[4]	Finkel T. Signal transduction by reactive oxygen species. J Cell Biol. 2011; 125: 376-393.
[5]	Janssen-Heininger YM, Mossman BT, Heintz NH, Forman HJ, Kalyanaraman B, et al. Redox-based regulation of signal transduction: principles, pitfalls, and promises. Free Radic Biol Med. 2008; 45: 1-17.
[6]	Navarro-Yepes J, Burns M, Anandhan A, Khalimonchuk O, Maria del Razo L, Quintanilla-Vega B, Pappa A, Panayiotidis MI, Franco R. Oxidative stress, redox signaling and authophagy: cell death versus survival. Antioxid Redox Signal. 2014; 21(1): 66-85.
[7]	Fischer R, Maier O. Interrelation of oxidative stress and inflammation in neurodegenerative diseases: role of TNF. Oxid Med Cell Longev. 2015; 2015: 610813.
[8]	Di Meo S, Reed TT, Venditti P, Victor VM. Role of ROS and RNS sources in physiological and pathological conditions. Oxid Med Cell Longev. 2016; 2016: 1245049.
[9]	Smeyne M, Smeyne RJ. Glutathione metabolism and Parkinson’s disease. Free Radical Biol Med. 2013; 62: 13-25.
[10]	Meister A, Anderson ME. Glutathione. Annu Rev Biochem. 1983; 52: 711-760.
[11]	Aoyama K, Nakaki T. Impaired glutathione synthesis in neurodegeneration. Int J Mol Sci. 2013; 14: 21021-21044.
[12]	Johnson WM, Wilson-Deflosse AL, Mieyal JJ. Dysregulation of glutathione homeostasis in neurodegenerative diseases. Nutrients. 2012; 4: 1399-1440.
[13]	Main PAE, Angley MT, O’Doherty CE,Thomas P, Fenech M. The potential role of the antioxidant and detoxification properties of glutathione in autism spectrum disorders: a systematic review and meta-analysis. Nutr Metab. (Lond) 2012; 9: 36.
[14]	Ma Q. Role of Nrf2 in oxidative stress and toxicity. Annu Rev Pharmacol Toxicol. 2013; 53: 301-426.
[15]	Huang Y, Li W, Su Z-Y, Kong A-NT. The complexity of the Nrf2 pathway: beyond the antioxidant response. J Nutr Biochem. 2015; 26: 1401-1413.
[16]	Tebay LE, Robertson H, Durant ST, Vitale SR, Penning TM, Dinkova-Kostova AT, Hayes JD. Mechanisms of activation of the transcription factor Nrf2 by redox stressor, nutrient cues, and energy status and the pathways through which it attenuates degenerative disease. Free Radical Biol Med. 2015; 88: 108-146.
[17]	Sandberg M, Patil J, D’Angelo B, Weber SG, Mallard C. Nrf2-regulation in brain health and disease: implication of cerebral inflammation. Neuropharmacol. 2014; 79: 298-306.
[18]	Huang Y, Li W, Su Z-Y, Kong A-N T. The complexity of the Nrf2 pathway: beyond the antioxidant response. J Nur Biochem. 2105; 26: 1401-1413.
[19]	Ramsey CP, Glass CA, Montgomery MB, Lindl KA, Ritson GP, Chia LA, et al. Expression of Nrf2 in neurodegenerative diseases. J Neuropathol Exp Neurol. 2007: 66: 75-85.
[20]	Marmiroli P, Cavaletti G. The glutamatertic neurotransmission in the central nervous system. Curr Med Chem. 2012; 19: 1269-1276.
[21]	Zhou Y, Danbolt NC. Glutamate as a neurotransmitter in the healthy brain. J Neural Transm. 2014; 121: 799-817.
[22]	Ohta S. Molecular hydrogen as a novel antioxidant. Overview of the advantages of hydrogen for medical applications. Meth Enzymol. 2015; 555: 289-317.
[23]	Nicolson GL, Ferreira de Mattos G, Settineri R, Costa C, Ellithorpe R, Rosenblatt S, La Valle J, Jimenez A, Ohta S. Clinical effects of hydrogen administration: from animal and human diseases to exercise medicine. Intern J Clin Med. 2016: 7: 32-76.
[24]	Settineri R, Ji J, Luo C, Ellithorpe RR, Ferreira de Mattos G, Rosenblatt S, La Valle J, Jinenez A, Ohta S, Nicolson GL. Effects of hydrogenized water in intracellular biomarkers for antioxidants, glucose uptake, insulin signaling and SIRT1 and telomerase activity. Am J Food Nutrit. 2016; 4: 161-168.
[25]	Navjot S, Singh R, Sarangi U, Saxena N, Chaudhary A, Kaur G, Kaul SC, Wadhwa R. Combinations of Ashwagandha leaf extracts lprotect brain-derived cells against oxidative stress and induced differentiation. PLos One. 2015; 10: e0120554.
[26]	Barayuga SM, Pang X, Andres MA, Panee A, Bellinger FP. Methamphetamine Decreases Levels of Glutathione Peroxidases 1 and 4 in SH-SY5Y Neuronal Cells: Protective Effects of Selenium. Neurotoxicol. 2013; 37: 240-246.
[27]	Lee J-M, Chan K, Kan YW, Johnson JA. Targeted disruption of Nrf2 causes regenerative immune-mediated hemolytic anemia. Proc Natl Acad Sci USA. 2004; 101: 9751-9756.
[28]	R Core Team. A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria, 2017. https://www.R-project.org.
[29]	Zhou Y, Danbolt NC. Glutamate as a neurotransmitter in the healthy brain. J Neural Transm. 2014; 121: 799-817.
[30]	Mayford M, Siegelbaum SA, Kandel ER. Synapses and memory storage. Cold Spring Har Perspect Biol. 2012; 4: a005751.
[31]	Ikonomidou C, Turski L. Why did NMDA receptor antagonists fail clinical trials for stroke and traumatic brain injury? Lancet Neurol. 2002; 1: 383-386.
[32]	Traynelis SF, Wollmuth LP, McBain CJ, Menniti FS, Vance KM, Ogden KK, et al. Glutamate receptor ion channels: structure, regulation and function. Pharmacol Rev. 2010; 62: 405-496.
[33]	Lewerenz J, Maher P. Chronic glutamate toxicity in neurodegenerative diseases-What is the evidence? Front Neurosci. 2015; 9: 469.
[34]	Pearce RK, Owen A, Daniel S, Jenner P, Marsden CD. Alterations in the distribution of glutathione in the substantia nigra in Parkinson’s disease. J Neural Transm. 1997; 104: 661-667.
[35]	Mischley LK, Conley KE, Shankland EG, Kavanagy TJ, Rosenfeld ME, Duda JE, White CC, Wilbur TK, De La Torre P, Padowski JM. Central nervous system uptake of intranasal glutathione in Parkinson’s disease. NPJ Parkinson's Dis. 2016; 2: 16002.
[36]	Sies H, Berndt C, Jones DP. Oxidative Stress. Annu Rev Biochem. 2017; 86: 715-748.
[37]	Liu Y, Hyde AS, Simpson MA, Barycki JJ. Emerging regulatory paradigms in glutathione metabolism. Adv Cancer Res. 2014; 122: 69-101.
[38]	Currais A, Maher P. Functional consequences of age-dependent changes in glutathione status in the brain. Antioxid Redox Signal. 2013; 19: 813-822.
[39]	Towsend DM, Tew KD. The role of glutathione-S-transferase in anti-cancer drug resistance. Oncogene 2003; 22: 7369-7375.
[40]	Suryakant KN, Khatri R, Jaiswal AK. Regulation of Nrf2 – An update. Free Radic Biol Med. 2014; 66.
[41]	Gao B, Doan A, Hybertson BM. The clinical potential of influencing Nrf2 signaling in degenerative and immunological disorders. Clin Pharmacol. 2014; 6: 19-39.
[42]	Su Z-Y, Shu L, Khor TO, Lee JH, Fuentes F, Kong A-N T. A perspective on dietary phytochemicals and cancer prevention: oxidative stress, Nrf2 and epigenomics. Top Curr Chem. 2013; 329: 133-162.
[43]	Ma Q, He X. Molecular basis of electrophilic and oxidative defense: promises and perils of Nrf2. Pharmacol Rev. 2012; 64: 1055-1081.
[44]	Loboda A, Damulewicz, Pyza E, Jozkowicz A, Dulak J. Role of Nrf2/HO-1 system in development, oxidative stress response and diseases: an evolutionarily conserved mechanism. Cell Mol Life Sci. 2016; 73: 3221-3247.
[45]	Zhang H, Davies KJA, Forman HJ. Oxidative stress response and Nrf2 signaling in aging. Free Radic Biol Med. 2015; 88: 314-336.