Enhancement mechanism of antioxidant enzyme gene expression by hydrogen molecules
© Kinjo et al.; licensee BioMed Central Ltd. 2013
Published: 4 December 2013
Redox regulation system protects our body from oxidative stress-injury and keeps redox homeostasis. The hydrogen molecules (H2) exist as stable gas in the ordinal temperature and atmosphere. Recent study reports H2 improve ischemia-reperfusion injury, glaucoma, Parkinson's disease and atherosclerosis of animal models. It is supposed from these improvement results that H2 participate in reduction of the oxidation stress, however, the reaction mechanism has not been clarified thoroughly. We surmised that intracellular redox regulation system is activated by H2 thereupon antioxidative activity is generated. Thus, we tried to find the effect of H2 on the Nrf2 pathway, one of the redox regulation systems.
Materials and methods
HT1080 cells, a human fibrosarcoma cell line, were incubated in a gas incubator at an atmosphere of 75%N2/20%O2/5%CO2 or 75%H2/20%O2/5%CO2 for 24 h. Then, after the cells were treated with H2O2 or fixative solution for 30 min or 15 min, the intracellular H2O2 and Nrf2 were determined by In cell analyzer and Confocal laser microscop using a BES-H2O2 or anti-Nrf2 antibody, respectively. Furthermore, after extraction of mRNA from the treated HT1080 cells, the gene expressions were examined by using Real-time PCR.
The quantity of intracellular H2O2 increased by hydrogen peroxide treatment was significantly decreased by pretreatment of H2. H2 enhanced the expression of catalase, glultathione peroxidase, Cu/Zn-superoxide dismutase, Nrf2 genes and Nrf2 protein.
It was suggested that H2 induced the expression level of antioxidant enzyme genes like catalase and glutathione peroxidase by increasing the expression level of the Nrf2 protein and decreased the amount of intracellular H2O2 induced by the H2O2 treatment in HT1080 cells.
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