Skip to main content

Treatment with Hydrogen-Rich Saline Delays Disease Progression in a Mouse Model of Amyotrophic Lateral Sclerosis

Abstract

Amyotrophic lateral sclerosis (ALS) is the most frequent adult-onset motor neuron disease, and accumulating evidence indicates that oxidative mechanisms contribute to ALS pathology, but classical antioxidants have not performed well in clinical trials. The aim of this work was to investigate the effect of treatment with hydrogen molecule on the development of disease in mutant SOD1 G93A transgenic mouse model of ALS. Treatment of mutant SOD1 G93A mice with hydrogen-rich saline (HRS, i.p.) significantly delayed disease onset and prolonged survival, and attenuated loss of motor neurons and suppressed microglial and glial activation. Treatment of mutant SOD1 G93A mice with HRS inhibited the release of mitochondrial apoptogenic factors and the subsequent activation of downstream caspase-3. Furthermore, treatment of mutant SOD1 G93A mice with HRS reduced levels of protein carbonyl and 3-nitrotyrosine, and suppressed formation of reactive oxygen species (ROS), peroxynitrite, and malondialdehyde. Treatment of mutant SOD1 G93A mice with HRS preserved mitochondrial function, marked by restored activities of Complex I and IV, reduced mitochondrial ROS formation and enhanced mitochondrial adenosine triphosphate synthesis. In conclusion, hydrogen molecule may be neuroprotective against ALS, possibly through abating oxidative and nitrosative stress and preserving mitochondrial function.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. 1.

    Bruijn LI, Miller TM, Cleveland DW (2004) Unraveling the mechanisms involved in motor neuron degeneration in ALS. Annu Rev Neurosci 27:723–749

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Cozzolino M, Ferri A, Carri MT (2008) Amyotrophic lateral sclerosis: from current developments in the laboratory to clinical implications. Antioxid Redox Signal 10:405–443

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Andersen PM, Al-Chalabi A (2011) Clinical genetics of amyotrophic lateral sclerosis: What do we really know? Nat Rev Neurol 7:603–615

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Rowland LP, Shneider NA (2001) Amyotrophic lateral sclerosis. N Engl J Med 344:1688–1700

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Miller RG, Rosenberg JA, Gelinas DF, Mitsumoto H, Newman D, Sufit R, Borasio GD, Bradley WG, Bromberg MB, Brooks BR, Kasarskis EJ, Munsat TL, Oppenheimer EA (1999) Practice parameter: the care of the patient with amyotrophic lateral sclerosis (an evidence-based review): report of the Quality Standards Subcommittee of the American Academy of Neurology: ALS Practice Parameters Task Force. Neurology 52:1311–1323

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Morrison KE (2002) Therapies in amyotrophic lateral sclerosis-beyond riluzole. Curr Opin Pharmacol 2:302–309

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Barber SC, Mead RJ, Shaw PJ (2006) Oxidative stress in ALS: a mechanism of neurodegeneration and a therapeutic target. Biochim Biophys Acta 1762:1051–1067

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Barber SC, Shaw PJ (2010) Oxidative stress in ALS: key role in motor neuron injury and therapeutic target. Free Radic Biol Med 48:629–641

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Fitzmaurice PS, Shaw IC, Kleiner HE, Miller RT, Monks TJ, Lau SS, Mitchell JD, Lynch PG (1996) Evidence for DNA damage in amyotrophic lateral sclerosis. Muscle Nerve 19:797–798

    CAS  PubMed  Google Scholar 

  10. 10.

    Andrus PK, Fleck TJ, Gurney ME, Hall ED (1998) Protein oxidative damage in a transgenic mouse model of familial amyotrophic lateral sclerosis. J Neurochem 71:2041–2048

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Pehar M, Beeson G, Beeson CC, Johnson JA, Vargas MR (2014) Mitochondria-targeted catalase reverts the neurotoxicity of hSOD1G 93A astrocytes without extending the survival of ALS-linked mutant hSOD1 mice. PLoS One 9:e103438

    Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Desnuelle C, Dib M, Garrel C, Favier A (2001) A double-blind, placebocontrolled randomized clinical trial of alpha-tocopherol (vitamin E) in the treatment of amyotrophic lateral sclerosis. ALS Riluzole-Tocopherol Study Group. Amyotroph Lateral Scler Other Motor Neuron Disord 2:9–18

    CAS  Article  PubMed  Google Scholar 

  13. 13.

    Orrell R, Lane R, Ross M (2007) Antioxidant treatment for amyotrophic lateral sclerosis/motor neuron disease. Cochrane Database Syst Rev CD002829

  14. 14.

    Graf M, Ecker D, Horowski R, Kramer B, Riederer P, Gerlach M, Hager C, Ludolph AC, Becker G, Osterhage J, Jost WH, Schrank B, Stein C, Kostopulos P, Lubik S, Wekwerth K, Dengler R, Troeger M, Wuerz A, Hoge A, Schrader C, Schimke N, Krampfl K, Petri S, Zierz S, Eger K, Neudecker S, Traufeller K, Sievert M, Neundörfer B, Hecht M, German vitamin E/ALS Study Group (2005) High dose vitamin E therapy in amyotrophic lateral sclerosis as add-on therapy to riluzole: results of a placebo-controlled double-blind study. J Neural Transm 112:649–660

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, Katsura K, Katayama Y, Asoh S, Ohta S (2007) Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13:688–694

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Ohno K, Ito M, Ichihara M, Ito M (2012) Molecular hydrogen as an emerging therapeutic medical gas for neurodegenerative and other diseases. Oxid Med Cell Longev 2012:353152

    Article  PubMed  PubMed Central  Google Scholar 

  17. 17.

    Fujita K, Seike T, Yutsudo N, Ohno M, Yamada H, Yamaguchi H, Sakumi K, Yamakawa Y, Kido MA, Takaki A, Katafuchi T, Tanaka Y, Nakabeppu Y, Noda M (2009) Hydrogen in drinking water reduces dopaminergic neuronal loss in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson’s disease. PLoS One 4:e7247

    Article  PubMed  PubMed Central  Google Scholar 

  18. 18.

    Fu Y, Ito M, Fujita Y, Ito M, Ichihara M, Masuda A, Suzuki Y, Maesawa S, Kajita Y, Hirayama M, Ohsawa I, Ohta S, Ohno K (2009) Molecular hydrogen is protective against 6-hydroxydopamine-induced nigrostriatal degeneration in a rat model of Parkinson’s disease. Neurosci Lett 453:81–85

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Li J, Wang C, Zhang JH, Cai JM, Cao YP, Sun XJ (2010) Hydrogen-rich saline improves memory function in a rat model of amyloid-beta-induced Alzheimer’s disease by reduction of oxidative stress. Brain Res 1328:152–161

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Wang C, Li J, Liu Q, Yang R, Zhang JH, Cao YP, Sun XJ (2011) Hydrogen-rich saline reduces oxidative stress and inflammation by inhibit of JNK and NF-κB activation in a rat model of amyloid-beta-induced Alzheimer’s disease. Neurosci Lett 491:127–132

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Wang H, Guan Y, Wang X, Smith K, Cormier K, Zhu S, Stavrovskaya IG, Huo C, Ferrante RJ, Kristal BS, Friedlander RM (2007) Nortriptyline delays disease onset in models of chronic neurodegeneration. Eur J Neurosci 26:633–641

    Article  PubMed  Google Scholar 

  22. 22.

    Elks CM, Mariappan N, Haque M, Guggilam A, Majid DS, Francis J (2009) Chronic NF-kB blockade reduces cytosolic and mitochondrial oxidative stress and attenuates renal injury and hypertension in SHR. Am J Physiol Renal Physiol 296:F298–F305

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Mariappan N, Soorappan RN, Haque M, Sriramula S, Francis J (2007) TNF-α-induced mitochondrial oxidative stress and cardiac dysfunction: restoration by superoxide dismutase mimetic Tempol. Am J Physiol Heart Circ Physiol 293:H2726–H2737

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Ferri A, Fiorenzo P, Nencini M, Cozzolino M, Pesaresi MG, Valle C, Sepe S, Moreno S, Carrì MT (2010) Glutaredoxin 2 prevents aggregation of mutant SOD1 in mitochondria and abolishes its toxicity. Hum Mol Genet 19:4529–4542

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Friedlander RM (2003) Apoptosis and caspases in neurodegenerative diseases. N Engl J Med 348:1365–1375

    CAS  Article  PubMed  Google Scholar 

  26. 26.

    Li M, Ona VO, Guégan C, Chen M, Jackson-Lewis V, Andrews LJ, Olszewski AJ, Stieg PE, Lee JP, Przedborski S, Friedlander RM (2000) Functional role of caspase-1 and caspase-3 in an ALS transgenic mouse model. Science 288:335–339

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Beckman JS, Carson M, Smith CD, Koppenol WH (1993) ALS, SOD and peroxynitrite. Nature 364:584

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Basso M, Samengo G, Nardo G, Massignan T, D’Alessandro G, Tartari S, Cantoni L, Marino M, Cheroni C, De Biasi S, Giordana MT, Strong MJ, Estevez AG, Salmona M, Bendotti C, Bonetto V (2009) Characterization of detergent-insoluble proteins in ALS indicates a causal link between nitrative stress and aggregation in pathogenesis. PLoS One 4:e8130

    Article  PubMed  PubMed Central  Google Scholar 

  29. 29.

    Casoni F, Basso M, Massignan T, Gianazza E, Cheroni C, Salmona M, Bendotti C, Bonetto V (2005) Protein nitration in a mouse model of familial amyotrophic lateral sclerosis: possible multifunctional role in the pathogenesis. J Biol Chem 280:16295–16304

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Chinta SJ, Andersen JK (2011) Nitrosylation and nitration of mitochondrial complex I in Parkinson’s disease. Free Radic Res 45:53–58

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Palacios-Callender M, Quintero M, Hollis VS, Springett RJ, Moncada S (2004) Endogenous NO regulates superoxide production at low oxygen concentrations by modifying the redox state of cytochrome c oxidase. Proc Natl Acad Sci U S A 101:7630–7635

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  32. 32.

    Sasaki S, Iwata M (2007) Mitochondrial alterations in the spinal cord of patients with sporadic amyotrophic lateral sclerosis. J Neuropathol Exp Neurol 66:10–16

    Article  PubMed  Google Scholar 

  33. 33.

    Kong J, Xu Z (1998) Massive mitochondrial degeneration in motor neurons triggers the onset of amyotrophic lateral sclerosis in mice expressing a mutant SOD1. J Neurosci 18:3241–3250

    CAS  PubMed  Google Scholar 

  34. 34.

    Cassina P, Cassina A, Pehar M, Castellanos R, Gandelman M, de León A, Robinson KM, Mason RP, Beckman JS, Barbeito L, Radi R (2008) Mitochondrial dysfunction in SOD1G93A-bearing astrocytes promotes motor neuron degeneration: prevention by mitochondrial-targeted antioxidants. J Neurosci 28:4115–4122

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Dupuis L, Gonzalez de Aguilar JL, Oudart H, de Tapia M, Barbeito L, Loeffler JP (2004) Mitochondria in amyotrophic lateral sclerosis: a trigger and a target. Neurodegener Dis 1:245–254

    Article  PubMed  Google Scholar 

  36. 36.

    Shi P, Gal J, Kwinter DM, Liu X, Zhu H (2010) Mitochondrial dysfunction in amyotrophic lateral sclerosis. Biochim Biophys Acta 1802:45–51

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  37. 37.

    Lipscomb GL, Schut GJ, Thorgersen MP, Nixon WJ, Kelly RM, Adams MW (2014) Engineering hydrogen gas production from formate in a hyperthermophile by heterologous production of an 18-subunit membrane-bound complex. J Biol Chem 289:2873–2879

    CAS  Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Shu-Yi Pan.

Ethics declarations

Conflict of interest

We declare that we have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Zhang, Y., Li, H., Yang, C. et al. Treatment with Hydrogen-Rich Saline Delays Disease Progression in a Mouse Model of Amyotrophic Lateral Sclerosis. Neurochem Res 41, 770–778 (2016). https://doi.org/10.1007/s11064-015-1750-7

Download citation

Keywords

  • Hydrogen molecule
  • Amyotrophic lateral sclerosis
  • Oxidative and nitrosative stress
  • Mitochondrial function