Hydrogen-rich saline promotes survival of retinal ganglion cells in a rat model of optic nerve crush

PLoS One. 2014 Jun 10;9(6):e99299. doi: 10.1371/journal.pone.0099299. eCollection 2014.


Objective: To investigate the effect of molecular hydrogen (H2) in a rat model subjected to optic nerve crush (ONC).

Methods: We tested the hypothesis that after optic nerve crush (ONC), retinal ganglion cell (RGC) could be protected by H₂. Rats in different groups received saline or hydrogen-rich saline every day for 14 days after ONC. Retinas from animals in each group underwent measurements of hematoxylin and eosin (H&E) staining, cholera toxin beta (CTB) tracing, gamma synuclein staining, and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining 2 weeks post operation. Flash visual evoked potentials (FVEP) and pupillary light reflex (PLR) were then tested to evaluate the function of optic nerve. The malondialdehyde (MDA) level in retina was evaluated.

Results: H&E, gamma synuclein staining and CTB tracing showed that the survival rate of RGCs in hydrogen saline-treated group was significantly higher than that in saline-treated group. Apoptosis of RGCs assessed by TUNEL staining were less observed in hydrogen saline-treated group. The MDA level in retina of H₂ group was much lower than that in placebo group. Furthermore, animals treated with hydrogen saline showed better function of optic nerve in assessments of FVEP and PLR.

Conclusion: These results demonstrated that H₂ protects RGCs and helps preserve the visual function after ONC and had a neuroprotective effect in a rat model subjected to ONC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Count
  • Cell Survival / drug effects
  • Disease Models, Animal
  • Evoked Potentials, Visual / drug effects
  • Hydrogen / pharmacology*
  • In Situ Nick-End Labeling
  • Light
  • Male
  • Malondialdehyde / metabolism
  • Nerve Crush*
  • Optic Nerve Injuries / pathology*
  • Optic Nerve Injuries / physiopathology
  • Pupil / drug effects
  • Pupil / radiation effects
  • Rats, Sprague-Dawley
  • Reflex / drug effects
  • Reflex / radiation effects
  • Retinal Ganglion Cells / drug effects
  • Retinal Ganglion Cells / pathology*
  • Sodium Chloride / pharmacology*
  • Staining and Labeling
  • gamma-Synuclein / metabolism


  • gamma-Synuclein
  • Sodium Chloride
  • Malondialdehyde
  • Hydrogen

Grant support

This work was supported by The National Basic Research Program of China (973 Program 2005CB724302) and the Creativity and Innovation Training Program of Second Military Medical University (MS2010035). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.