Effects of hydrogen-rich water on aging periodontal tissues

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OPEN SUBJECT AREAS: INFLAMMASOME PRE-CLINICAL STUDIES Received 3 February 2014 Accepted 13 June 2014 Published 2 July 2014 Correspondence and requests for materials should be addressed to T.T. (tomofu@md. okayama-u.ac.jp) Effects of hydrogen-rich water on aging periodontal tissues in rats Takaaki Tomofuji, Yuya Kawabata, Kenta Kasuyama, Yasumasa Endo, Toshiki Yoneda, Mayu Yamane, Tetsuji Azuma, Daisuke Ekuni & Manabu Morita Department of Preventive Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5- 1 Shikata-cho, Kita-ku, Okayama 700-8558, Japan. Oxidative damage is involved in age-related inflammatory reactions. The anti-oxidative effects of hydrogen-rich water suppress oxidative damage, which may aid in inhibiting age-related inflammatory reactions. We investigated the effects of drinking hydrogen-rich water on aging periodontal tissues in healthy rats. Four-month-old male Fischer 344 rats (n 5 12) were divided into two groups: the experimental group (hydrogen-rich water treatment) and the control group (distilled water treatment). The rats consumed hydrogen-rich water or distilled water until 16 months of age. The experimental group exhibited lower periodontal oxidative damage at 16 months of age than the control group. Although protein expression of interleukin-1b did not differ, gene expression of Nod-like receptor protein 3 inflammasomes was activated in periodontal tissues from the experimental group as compared with the control group. Drinking hydrogen-rich water is proposed to have anti-aging effects on periodontal oxidative damage, but not on inflammatory reactions in healthy rats. Aging is a reality for all living organisms, and is the result of the progressive accumulation of deleterious changes that reduce an organism’s ability to resist stress, decreasing the possibility of survival. One of the most popular theories used to explain the mechanisms of aging is the mitochondrial free radical theory of aging1,2. In this theory, mitochondrial function declines in an age-dependent manner, thereby enhancing the production of reactive oxygen species (ROS)3. Moreover, during aging, it is reported that increased production of ROS induces oxidative mitochondrial DNA (mtDNA) damage, which can stimulate the activation of the Nod-like receptor protein (NLRP) 3 inflammasome in tissues4. Because NLRP3 inflammasomes serve as a platform for caspase-1 activation and subsequent proteolytic maturation of the potent pro-inflammatory cytokine interleukin (IL)-1b4,5, these indicate that oxidative mtDNA damage can stimulate age-related inflammatory reactions in tissues. Therefore, reduction in oxidative mtDNA damage through antioxidant therapy may be effective in suppressing age-related inflammatory reactions. Molecular hydrogen is considered a novel antioxidant that can reduce oxidative damage6. Drinking hydrogen-rich water is an alternative mode of molecular hydrogen delivery. Clinical and animal studies have demonstrated that drinking hydrogen-rich water provides numerous health benefits, such as in regards to the serum lipid profiles of those at risk for metabolic syndrome7,8, quality of life after radiation exposure9, oxidative damage in patients with chronic hepatitis B10 and nonalcoholic steatohepatitis related hepatocarcinogenesis11. In a previous study, we also found that drinking hydrogen-rich water suppressed ligature-induced periodontal inflammation in a rat model12. Notably, these studies mainly focused on the effects of hydrogen-rich water on human disease status and animal disease models. However, it remains unclear what effects drinking hydrogen-rich water have over the course of a lifetime. In the present study, we hypothesized that drinking hydrogen-rich water might suppress age-related oxidative mtDNA damage and inflammatory reactions in the periodontal tissue. Thus, the purpose of this study was to investigate the anti-aging effects of drinking hydrogen-rich water on oxidative mtDNA damage and inflammat- ory reactions in rat periodontal tissue. The main outcomes of this study were oxidative mtDNA damage and IL-1b expression in the periodontal tissue. The level of oxidative damage was determined by measuring 8-hydroxy- deoxyguanosine (8-OHdG), which is highly specific for DNA damage13. To gain better insight into the mech- anism of action of hydrogen-rich water, we analyzed histological changes and NLRP3 inflammasome activation. Results Body weights and food consumption. Body weights (mean 6 SD of 6 rats, g) of the baseline, control and experimental groups were 286 6 10, 324 6 28 and 346 6 16, respectively. There were no significant differences SCIENTIFIC REPORTS | 4 : 5534 | DOI: 10.1038/srep05534 1

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