Effects of Hydrogenized Water on Intracellular Biomarkers for Antioxidants, Glucose Uptake, Insulin Signaling and SIRT 1 and Telomerase Activity

Abstract

Hydrogen has been shown in several clinical trials to be completely safe without adverse events and there are no warnings in the literature of its toxicity or adverse effects during long-term exposure. Molecular hydrogen has proven useful and convenient as a novel antioxidant and modifier of gene expression in many conditions where oxidative stress and changes in gene expression result in cellular damage. Our intracellular biomarker studies have shown that a hydrogenized water drink formula containing 2.6 ppm dissolved hydrogen was able to penetrate cellular membranes and function as an antioxidant in human liver cells (HePG2) utilizing the Cellular Antioxidant Assay (CAA). This assay uses the protection of a florescent probe as a marker for cellular damage by reactive oxygen species (ROS), such as peroxyl radical, and compares this to the known antioxidant standard, Quercetin. Using this system oxidative damage was reduced in a dose-dependent manner. One ml of hydrogenized water was found to possess antioxidant capacity equivalent to 0.05 µmole of quercetin. When examined in a human colon cell line (Caco-2 cells), hydrogenized water demonstrated a dose-and time-dependent permeability inhibition of an intracellular fluorescent glucose derivative (2-NBDG), indicating decreased glucose uptake. In another study, the impact of hydrogenized water on Akt phosphorylation (Ser473), a biomarker for insulin signaling, was monitored in human skeletal muscle cells. The hydrogenized water treatment markedly elevated the level of phosphorylation of Akt (Ser473) in a dose-dependent manner. The anti-aging effects of hydrogenized water were examined utilizing SIRT1 expression as a biomarker of aging in human umbilical cells (HUVECs). Hydrogenized water increased dose-dependent SIRT1 gene expression. Hydrogenized water also increased telomerase activity (an anti-aging biomarker in HUVEC cells) up to 148% when cells were treated with media containing 25% hydrogenized water formula. Increased telomerase activity caused by hydrogenized water may be able to protect telomeres from degradation, suggesting the possible use of hydrogenized water in therapeutic interventions of age-related diseases. These studies show that commercial hydrogenized water improved the levels or activities of a few intracellular biomarkers specific for antioxidant activity, glucose uptake, insulin signaling and SIRT 1 and telomerase activities. Industrial Relevance: The molecular hydrogen used in this study indicates that certain commercial sources of hydrogenized water can provide similar antioxidant and gene expression modifications seen in other sources of molecular hydrogen. The biomarkers evaluated here lend well to hydrogenized water's biological activity relating to health conditions and aging.

Hydrogenized Water Effects on Protection of Brain Cells from Oxidative Stress and Glutamate Toxicity

“…At the time of initial packaging, the infused hydrogen within the test water was analyzed by both manometric and diaphragm polarographic electrode measurements at final concentration of 2.6 ppm (1.3 mM H 2 ) and pH 7.0. The Izumio hydrogenized water was used in all of the assays [24].…”

“…Human neuroblastoma SH-SY5Y cells were cultivated in a 1:1 mixture of Eagle's Minimum Essential Medium and F12 Medium with fetal bovine serum at a final concentration of 10%, seeded into a 96-well cell culture plate and cultured for 24 hr before addition of the test compound in cell culture media. When media containing hydrogenized water was used, culture medium dry powder was suspended and dissolved in freshly opened hydrogenized water, and different final concentrations of the hydrogenized water in cell culture grade water were used at v/v 1.00, 0.50, 0.250, 0.0125, 0.00625 and 0.003125 of hydrogenized water to cell culture grade water, as previously used for other cell types [24]. Culture medium was then sterile-filtered and used to treat neuronal cells immediately.…”

“…Hydrogenized water has been shown to act as a cellular antioxidant and gene regulator [22,23]. Previously we found that hydrogenized water was an effective cellular antioxidant and could decrease transport of glucose in a dose-and time-dependent manner and elevate the levels of phosphorylation of Akt, a protein kinase B biomarker for insulin signaling [24]. We also found that hydrogenized water could increase the levels of age-related SIRT1 gene expression and increase the expression of telomerase activity in a dose-dependent process [24].…”

“…Previously we found that hydrogenized water was an effective cellular antioxidant and could decrease transport of glucose in a dose-and time-dependent manner and elevate the levels of phosphorylation of Akt, a protein kinase B biomarker for insulin signaling [24]. We also found that hydrogenized water could increase the levels of age-related SIRT1 gene expression and increase the expression of telomerase activity in a dose-dependent process [24]. Here we examined the ability of hydrogenized water in vitro to protect brain cells from oxidative damage by increasing glutathione levels and reducing glutamate toxicity.…”

Molecular Hydrogen as Medicine: An Assessment of Administration Methods

“…Much of the clinical research to date has focused on the effects of HRW on oxidative stress in conditions ranging from age-related diseases to exercise fatigue [52][53][54][55]. HRW is versatile option for the treatment of many ailments, as it is portable and can be produced in a number of ways.…”