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The Evaluation and Quantitation of H2 Gas Metabolism in Rats

Radomir Hyspler1 *, Alena Ticha1 , Henk Schierbeek2 , Alexander Galkin3 , Zdenek Zadak1 1 Department of Research and Development, University Hospital Hradec Kralove, Hradec Kralove, Czech Republic, 2 Division of Mother and Child, University of Amsterdam, Amsterdam, Netherlands, 3 School of Biological Sciences, Queen’s University, Belfast, United Kingdom



Despite the significant interest in molecular hydrogen as an antioxidant in the last eight years, its quantitative metabolic parameters in vivo are still lacking, as is an appropriate method for determination of hydrogen effectivity in the mammalian organism under various conditions.

Basic Procedures

Intraperitoneally-applied deuterium gas was used as a metabolic tracer and deuterium enrichment was determined in the body water pool. Also, in vitro experiments were performed using bovine heart submitochondrial particles to evaluate superoxide formation in Complex I of the respiratory chain.

Main Findings

A significant oxidation of about 10% of the applied dose was found under physiological conditions in rats, proving its antioxidant properties. Hypoxia or endotoxin application did not exert any effect, whilst pure oxygen inhalation reduced deuterium oxidation. During in vitro experiments, a significant reduction of superoxide formation by Complex I of the respiratory chain was found under the influence of hydrogen. The possible molecular mechanisms of the beneficial effects of hydrogen are discussed, with an emphasis on the role of iron sulphur clusters in reactive oxygen species generation and on iron species-dihydrogen interaction.

Principal Conclusions

According to our findings, hydrogen may be an efficient, non-toxic, highly bioavailable and low-cost antioxidant supplement for patients with pathological conditions involving ROSinduced oxidative stress.