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. 2002 Nov;40(1-3):139–149. doi: 10.1023/A:1023936421448

Protective mechanism of reduced water against alloxan-induced pancreatic β-cell damage: Scavenging effect against reactive oxygen species

Yuping Li 1, Tomohiro Nishimura 1, Kiichiro Teruya 1, Tei Maki 1, Takaaki Komatsu 1, Takeki Hamasaki 1, Taichi Kashiwagi 1, Shigeru Kabayama 2, Sun-Yup Shim 1, Yoshinori Katakura 1, Kazuhiro Osada 1, Takeshi Kawahara 1, Kazumichi Otsubo 2, Shinkatsu Morisawa 2, Yoshitoki Ishii 3, Zbigniew Gadek 4, Sanetaka Shirahata 1,✉
PMCID: PMC3449533  PMID: 19003114

Abstract

Reactive oxygen species (ROS) cause irreversible damage to biological macromolecules, resulting in many diseases. Reduced water (RW) such as hydrogen-rich electrolyzed reduced water and natural reduced waters like Hita Tenryosui water in Japan and Nordenau water in Germany that are known to improve various diseases, could protect a hamster pancreatic β cell line, HIT-T15 from alloxan-induced cell damage. Alloxan, a diabetogenic compound, is used to induce type 1 diabetes mellitus in animals. Its diabetogenic effect is exerted via the production of ROS. Alloxan-treated HIT-T15 cells exhibited lowered viability, increased intracellular ROS levels, elevated cytosolic free Ca2+ concentration, DNA fragmentation, decreased intracellular ATP levels and lowering of glucose-stimulated release of insulin. RW completely prevented the generation of alloxan-induced ROS, increase of cytosolic Ca2+ concentration, decrease of intracellular ATP level, and lowering of glucose-stimulated insulin release, and strongly blocked DNA fragmentation, partially suppressing the lowering of viability of alloxan-treated cells. Intracellular ATP levels and glucose-stimulated insulin secretion were increased by RW to 2–3.5 times and 2–4 times, respectively, suggesting that RW enhances the glucose-sensitivity and glucose response of β-cells. The protective activity of RW was stable at 4 °C for over a month, but was lost by autoclaving. These results suggest that RW protects pancreatic β-cells from alloxan-induced cell damage by preventing alloxan-derived ROS generation. RW may be useful in preventing alloxan-induced type 1-diabetes mellitus.

Keywords: alloxan, apoptosis, diabetes, HIT-T15 cells, insulin, pancreatic beta cells, reactive oxygen species, reduced water

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