Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals

Abstract

Acute oxidative stress induced by ischemia-reperfusion or inflammation causes serious damage to tissues, and persistent oxidative stress is accepted as one of the causes of many common diseases including cancer. We show here that hydrogen (H2) has potential as an antioxidant in preventive and therapeutic applications. We induced acute oxidative stress in cultured cells by three independent methods. H2 selectively reduced the hydroxyl radical, the most cytotoxic of reactive oxygen species (ROS), and effectively protected cells; however, H2 did not react with other ROS, which possess physiological roles. We used an acute rat model in which oxidative stress damage was induced in the brain by focal ischemia and reperfusion. The inhalation of H2 gas markedly suppressed brain injury by buffering the effects of oxidative stress. Thus H2 can be used as an effective antioxidant therapy; owing to its ability to rapidly diffuse across membranes, it can reach and react with cytotoxic ROS and thus protect against oxidative damage.

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Figure 1: Molecular hydrogen dissolved in medium selectively reduces hydroxyl radicals in cultured cells.
Figure 2: Molecular hydrogen protects cultured PC12 cells by scavenging hydroxyl radicals.
Figure 3: Spin-trapping identifies the free radical species that H2 reduces.
Figure 4: Molecular hydrogen dissolved in solution scavenges hydroxyl radicals at 23 °C and pH 7.4 in cell-free systems.
Figure 5: Inhalation of hydrogen gas protects against ischemia-reperfusion injury.
Figure 6: Inhalation of H2 gas improved brain injury after 1 week.

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Acknowledgements

This work was supported by grants to S.O. from the Ministry of Health, Labor and Welfare (H17-Chouju-009, longevity science; and 17A-10, nervous and mental disorders) and the Ministry of Education, Culture, Sports, Science and Technology (16390257).

Author information

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Authors

Contributions

S.O. conceived the experiments. S.O., I.O., K.K. and Y.K. designed the experiments. I.O., S.A. and S.O. performed data analysis. I.O., M.I., K.T., M.W., K.N, K.Y., S.A. and S.O. performed the experiments. S.O. and I.O. wrote the paper.

Corresponding author

Correspondence to Shigeo Ohta.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Molecular hydrogen dissolved in culture medium does not reduce cellular hydrogen peroxide and nitric oxide. (PDF 351 kb)

Supplementary Fig. 2

pH, H2 and O2 maintain constant in culture medium in a closed flask filled with a mixed gas. (PDF 75 kb)

Supplementary Fig. 3

Several methods confirm protection of cells by H2 against oxidative stress. (PDF 470 kb)

Supplementary Fig. 4

Molecular hydrogen protects cultured neurons from ischemia and reperfusion in vitro. (PDF 553 kb)

Supplementary Fig. 5

Cerebral blood flow is not influenced by H2 inhalation. (PDF 417 kb)

Supplementary Fig. 6

The brain after induction of ischemia reperfusion injury with or without H2 treatment was immunostained. (PDF 305 kb)

Supplementary Table 1

Physiological parameters during cerebral ischemia reperfusion (PDF 255 kb)

Supplementary Methods (PDF 140 kb)

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Ohsawa, I., Ishikawa, M., Takahashi, K. et al. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13, 688–694 (2007). https://doi.org/10.1038/nm1577

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