Hydrogen-Rich Water and Heme Oxygenase Modulators

HRW was prepared by using a hydrogen water generator from Hydrogen (Osmo-star Soriano S.L., Alicante, Spain), that uses electrolysis method to break down H₂O into H₂. HRW was intraperitoneally injected 1 h before testing at a concentration of 0.3 mM. Tin protoporphyrin IX (SnPP) and CoPP were acquired from Frontier Scientific (Livchem GmbH & Co., Frankfurt, Germany), and dicoumarol was purchased from Eurodiagnostico S.L. (Madrid, Spain). All of them were dissolved in dimethyl sulfoxide (DMSO, 1% solution in saline (0.4% NaCl). Glibenclamide purchased from Sigma–Aldrich (St. Louis, MO, USA) was dissolved in saline solution. SnPP, Glibenclamide and dicoumarol were administered 30 min before testing at a dose of 10 mg/kg, and CoPP at 3 h prior to the tests at dose of 2.5 mg/kg according to earliest studies [13 (link),31 (link)]. All drugs were prepared right before their use and were intraperitoneally injected in a final volume of 10 mL/kg. For each treatment, the respective control group received the corresponding vehicle (VEH).