Hydrogen-rich pure water prevents superoxide formation in brain slices of vitamin C-depleted SMP30/GNL knockout mice

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Abstract

Hydrogen is an established anti-oxidant that prevents acute oxidative stress. To clarify the mechanism of hydrogen’s effect in the brain, we administered hydrogen-rich pure water (H2) to senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice, which cannot synthesize vitamin C (VC), also a well-known anti-oxidant. These KO mice were divided into three groups; recipients of H2, VC, or pure water (H2O), administered for 33 days. VC levels in H2 and H2O groups were <6% of those in the VC group. Subsequently, superoxide formation during hypoxia-reoxygenation treatment of brain slices from these groups was estimated by a real-time biography imaging system, which models living brain tissues, with Lucigenin used as chemiluminescence probe for superoxide. A significant 27.2% less superoxide formed in the H2 group subjected to ischemia–reperfusion than in the H2O group. Thus hydrogen-rich pure water acts as an anti-oxidant in the brain slices and prevents superoxide formation.

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Materials and methods

Hydrogen-rich pure water. Pure water was produced by the following processes: passage through (1) a reverse osmosis/ultrafiltration unit, (2) an ion-exchange resin, and (3) an ultrafiltration membrane (pure water: pH 6.9 ± 0.05; electric conductivity 0.7 ± 0.2 μS/cm). Hydrogen-rich pure water then resulted from dissolving hydrogen gas directly into pure water and had the following physical properties: pH 6.7 ± 0.1, low electric conductivity (0.9 ± 0.2 μS/cm), high content of dissolved hydrogen (1.2 ± 0.1 

Effect of hydrogen-rich pure water on body weight

SMP30/GNL KO mice were divided into three groups, mice fed hydrogen-rich pure water (H2), VC water (VC), or pure water (H2O) after weaning at 30 days of age. To investigate the effect of H2, VC, or H2O administration on growth, we compared body weight changes (Fig. 1). All three groups of SMP30/GNL KO mice gained the same amount of weight throughout the experiment. That is, the body weights of H2, VC, and H2O administration groups at 63 days of age were 26.1 ± 0.5, 25.0 ± 1.1, and 24.9 ± 0.9 g,

Discussion

In this study, we demonstrated that the administration of hydrogen-rich pure water created a pronounced decrease of superoxide formation in brain slices. These brain tissues came from VC-negative SMP30/GNL KO mice and were examined during hypoxia-reoxygenation treatment by using a real-time biographic system in which Lucigenin functioned as a chemiluminescence probe that detects superoxide. This outcome reflects a decrease of ROS generation in brain slices during ischemia and coincides with the

Acknowledgments

This study is supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture, Japan (to A.I., S.H. and N.S.), and a Grant-in-Aid for Smoking Research Foundation, Japan (to A.I.). We thank Ms. P. Minick for the excellent English editorial assistance. Vitamin C powder was kindly provided by DSM Nutrition Japan.

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    In the current study, we investigated whether chronic intake of GeO2 results in cochlear mitochondrial impairment and associated SNHL in CBA/J mice. Next, we investigated the effects of ROS scavengers, taurine, coenzyme Q10 (CoQ10), or hydrogen-rich water (Huxtable, 1992; Erdem et al., 2000; Qiao et al., 2015; Koh et al., 2014; Das et al., 2009; Manna et al., 2009; Alam and Hafiz, 2011; Roy and Sil, 2012; M Sikorska et al., 2014; M Sikorska et al., 2014; Sohet et al., 2009; Someya et al., 2009; Yamada et al., 2015; Ohsawa et al., 2008; Sato et al., 2008; Ohsawa et al., 2007; Hayashida et al., 2008; Yoshida et al., 2012; Fukuda et al., 2007; Nakashima-Kamimura et al., 2009; Lin et al., 2011; Fransson et al., 2021), on cochlear degeneration and SNHL induced by GeO2. Female CBA/J mice were purchased from CLEA Japan (Tokyo, Japan).

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These authors contributed equally to this work.

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