Hydrogen Treatment Protects against Cell Death Senescence

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J. Microbiol. Biotechnol. (2017), 27(2), 365–371 https://doi.org/10.4014/jmb.1608.08011 Review Research Article jmb Hydrogen Treatment Protects against Cell Death and Senescence Induced by Oxidative Damage A Lum Han1, Seong-Hoon Park2*, and Mi Sung Park3 Department of Family Medicine, Medical Hospital, Wonkwang University, Iksan 54538, Republic of Korea Department of Radiology Medicine, Medical Hospital, Wonkwang University, Iksan 54538, Republic of Korea Institute for Metabolic Disease, School of Medicine, Wonkwang University, Iksan 54538, Republic of Korea Received: August 5, 2016 Revised: October 17, 2016 Accepted: October 24, 2016 First published online October 25, 2016 *Corresponding author Phone: +82-63-859-5890; Fax: +82-63-850-6799; E-mail: shpark99@wku.ac.kr pISSN 1017-7825, eISSN 1738-8872 Copyright© 2017 by The Korean Society for Microbiology and Biotechnology Hydrogen has potential for preventive and therapeutic applications as an antioxidant. However, micro- and macroparticles of hydrogen in water disappear easily over time. In order to eliminate reactive oxygen species (ROS) related with the aging process, we used functional water containing nanoparticle hydrogen. Nanoparticle hydrogen does not disappear easily and collapse under water after long periods of time. We used murine embryonic fibroblasts that were isolated from 12.5-day embryos of C57BL/6 mice. We investigated the ability of nanoparticle hydrogen in water to suppress hydroxyurea-induced ROS production, cytotoxicity, and the accumulation of β-galactosidase (an indicator of aging), and promote cell proliferation. The accumulation of β-galactosidase in the cytoplasm and the appearance of abnormal nuclei were inhibited by daily treatment of cells with hydrogen water. When the aging process was accelerated by hydroxyurea-induced oxidative stress, the effect of hydrogen water was even more remarkable. Thus, this study showed the antioxidant and anti- senescence effects of hydrogen water. Nanoparticle hydrogen water is potentially a potent anti-aging agent. Keywords: Hydrogen-rich water, hydrogen nanoparticles, anti-aging, antioxidant Introduction Reactive oxygen species (ROS) include oxygen-derived free radicals such as hydroxyl radicals ( OH), superoxide anions (O ), alkoxyl radicals (RO ), and peroxyl radicals (RO ), as well as oxygen-derived non-radical species such as hydrogen peroxide (H O ) [1]. As major ROS generators, mitochondria are constantly exposed to high levels of ROS, with detrimental consequences, including oxidative damage to mitochondrial DNA [2]. Thus, increased O and OH associated with mitochondrial DNA damage have been implicated in cellular apoptosis [3]. The deleterious role of free radicals in aging and senescence was first suggested by Dr. Denham Harman in 1956 [4]. Sixty years after his publication, numerous studies have provided evidence supporting the central hypothesis, and various reviews on free radicals and aging have been published [5- 8]. Hydrogen, the most abundant and lightest chemical element in nature, has anti-apoptotic, antioxidant, anti- inflammatory, and anti-allergy effects, and thus can be used to treat various diseases [9]. Molecular hydrogen diffuses rapidly into cells and tissues, and selectively reduces OH and peroxynitrite anions, which are the most cytotoxic ROS species. The effects of hydrogen occur without affecting normal ROS signaling or disrupting normal metabolic redox reactions that influence cell signaling [10]. Hydrogen is combustible in air at concentrations between 4% and 75%. However, in water, hydrogen is a stable gas that can only react with oxide radical ions ( O ) and OH at low reaction rates [11]. The reaction rate constants of O and OH with other biomolecules are generally in the order of 10 - 10 M ·s , whereas those with hydrogen are in the order of 10 M ·s . Nevertheless, hydrogen, being a small molecule that can easily distribute throughout cells and the body, is expected to have high collision rates with other February 2017 ⎪ Vol. 27 ⎪ No. 2 • − •• − 1− 1− 7 1−1− 01 9 •• 2 −• • • 2 −• • 22 2 • 3 2 1

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