Inhaled hydrogen gas therapy for prevention of testicular ischemia/reperfusion injury in rats

J Pediatr Surg. 2012 Apr;47(4):736-42. doi: 10.1016/j.jpedsurg.2011.09.035.


Purpose: This study evaluated whether 2% hydrogen (H(2)) gas therapy protects against testicular ischemia/reperfusion injury which results in increased formation of reactive oxygen species and/or reactive nitrogen species, leading to testicular apoptosis and impaired spermatogenesis.

Methods: Pubertal six-week-old Spraque-Dawley rats were assigned to 5 groups (10 animals/group) as follows: group A was a sham operated group; groups B, C, D, and E underwent 5 hours of left testicular ischemia followed by 0, 30, 60, and 120 minutes of 2% H(2) gas therapy, respectively. Histological analysis was performed to verify structure and morphology of the testes and to investigate Johnsen scores, mean seminiferous tubule diameter, and the number of germ cell layers to classify spermatogenesis. Germ cell apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling assay and Bax/Bcl-2 ratio real-time polymerase chain reaction. We also investigated malondialdehyde levels as an indicator of lipid peroxidation.

Results: Compared to the sham group (A), germ cell apoptosis and lipid peroxidation in the ischemia group (B) were significantly increased with abnormal morphology and impaired spermatogenesis. In contrast, amelioration of testicular damages was evident in the H(2) therapy groups (C, D, and E).

Conclusions: Our results showed that inhalation of 2% H(2) gas may be a promising therapy with anti-apoptotic and anti-oxidant properties in cases of testicular ischemia/reperfusion injury.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Inhalation
  • Animals
  • Antioxidants / pharmacology
  • Antioxidants / therapeutic use*
  • Apoptosis / drug effects
  • Biomarkers / metabolism
  • Disease Models, Animal
  • Hydrogen / pharmacology
  • Hydrogen / therapeutic use*
  • Lipid Peroxidation / drug effects
  • Male
  • Malondialdehyde / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / etiology
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / prevention & control*
  • Spermatic Cord Torsion / complications*
  • Spermatogenesis / drug effects
  • Treatment Outcome
  • bcl-2-Associated X Protein / metabolism


  • Antioxidants
  • Bax protein, rat
  • Biomarkers
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Malondialdehyde
  • Hydrogen