Hydrogen-rich saline attenuates skin ischemia/reperfusion induced apoptosis via regulating Bax/Bcl-2 ratio and ASK-1/JNK pathway

J Plast Reconstr Aesthet Surg. 2015 Jul;68(7):e147-56. doi: 10.1016/j.bjps.2015.03.001. Epub 2015 Mar 19.


Introduction: Many pathways have been reported involving the effect of hydrogen-rich saline on protecting skin flap partial necrosis induced by the inflammation of ischemia/reperfusion injury. This study focused on the influence of hydrogen-rich saline treatment on apoptosis pathway of ASK-1/JNK and Bcl-2/Bax radio in I/R injury of skin flaps.

Methods: Adult male Sprague-Dawley rats were divided into three groups. Group 1 was sham surgery group, Group 2 and 3 were ischemia/reperfusion surgery treated with physiological saline and hydrogen-rich saline respectively. Blood perfusion of flap was measured by Laser doppler flowmeters. Hematoxylin and eosin staining was used to observe morphological changes. Early apoptosis in skin flap was observed through TUNEL staining and presented as the percentage of TUNEL-positive cells of total cells. pASK-1, pJNK, Bcl-2 and Bax were examined by immunodetection. In addition Bcl-2, Bax and caspase-3 were detected by qPCR. Caspase-3 activity was also measured.

Results: Compared to the Group 2, tissues from the group 3 were observed with a high expression of Bcl-2 and a low expression of pASK-1, pJNK, and Bax, a larger survival area and a high level of blood perfusion. Hydrogen-rich saline ameliorated inflammatory infiltration and decreased cell apoptosis.

Conclusion: The results indicate that hydrogen-rich saline could ameliorate ischemia/reperfusion injury and improve flap survival rate by inhibiting the apoptosis factor and, at the same time, promoting the expression of anti-apoptosis factor.

Keywords: Apoptosis; Hydrogen-rich saline; Ischemia/reperfusion; Skin flaps.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Apoptosis / genetics
  • Free Tissue Flaps / blood supply
  • Hydrogen / administration & dosage*
  • Ischemia / prevention & control*
  • MAP Kinase Kinase Kinase 5 / metabolism
  • Male
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / prevention & control*
  • Saline Solution, Hypertonic / administration & dosage*
  • Skin / blood supply*
  • bcl-2-Associated X Protein / metabolism


  • Proto-Oncogene Proteins c-bcl-2
  • Saline Solution, Hypertonic
  • bcl-2-Associated X Protein
  • Hydrogen
  • MAP Kinase Kinase Kinase 5
  • MAP3K5 protein, human