Abstract

Reaclive oxygen species (ROS) cause irreversible damage to biologica] macromolecules, resulting in many dis- eases. Reduced water (RW) such as hydrogen-n'ch electrolyzed reduced water and natura] reduced waters like Hita Tenryosui water in Japan and Nordenau water in Germany thai are known to improve various diseases, could protecl a hamster pancreatic B cell line, HIT-TIS from alloxan-induced cell damage. Alloxan, a diabelogenic compound, is used to induce type 1 diabeles mellitus in animals. Its diabelogenic effect is exerted via the pro- duction of ROS. Alloxan-trealed HIT-TIS cells exhibited lowered viability, increased intracellular ROS levels, elevated cylosolic free Ca2+ concentration, DNA fragmentation, decreased intracellular A'I‘ P levels and lowering of glucose-sl'unulated release of insulin. RW completely prevemed the generation of alloxan-induced Ro8, increase of cytosolic Ca2+ oonoentration. decrease of intracellular ATP level, and lowering of glucose-sl'unulated insulin release, and strongly blocked DNA fragmentation. panially suppressing the lowen'ng of viability of alloxan-treated cells. lmracellular ATP levels and glucose-stimulated insulin secretion were increased by RW to 2-35 times and 2-4 times, respectively, suggesting thal RW enhances the glucose-sensitivity and glucose response of fi-cells. The protective activity of RW was stable al 4 °C for aver a month, but was lost by auloclaving. These results suggesl that RW protects pancreatic fl-cells from alloxan-induced cell damage by preventing alloxan-derived ROS generation. RW may be useful in preventing alloxan-induced type 1-diabetes mellitus. Abbreviations: BSA, bovine serum albumin; DCF, dichlorofiurescein; DCFH-DA, 2’,7’-dichlorofluorescin- diacetate; ELISA, enzyme-linked immunosorbenl assay; ERW. electnolyzed-reduced water; HBSS. Hank’s balanoed sall solution; IDDM, insulin-dependem diabetes mellitus; IlDM, insulin-independem diabetes mellitus; KRB, Krebs Ringer bicarbonate buffer; NRW. nature reduced water; ROS, reactive oxygen species; RW, reduced water.