Hydrogen-rich medium protects human skin fibroblasts from high glucose or mannitol induced oxidative damage

Biochem Biophys Res Commun. 2011 Jun 3;409(2):350-5. doi: 10.1016/j.bbrc.2011.05.024. Epub 2011 May 8.


Reactive oxygen species (ROS) are an important factor in the development of skin lesions in diabetes. A new antioxidant, hydrogen, can selectively neutralize hydroxyl radicals (()OH) and peroxynitrite (ONOO(-)) in cell-free systems, whereas it seldom reacts with other ROS. Fibroblasts are a key component of skin. In the present study, we investigated the protective effects of hydrogen-rich medium on human skin fibroblasts (HSFs) under oxidative stress. Confocal microscopy was used to assay both the intracellular superoxide anion (O(2)(-)) concentration and the mitochondrial membrane potential (ΔΨ). Cell viability was determined using the Cell Counting Kit-8 (CCK-8). The concentrations of cellular malonaldehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) were also measured. The results revealed that both mannitol and high glucose could cause oxidative stress in HSFs. Interestingly, the use of a hydrogen-rich medium significantly reduced the level of intracellular O(2)(-), stabilized the ΔΨ and attenuated production of MDA, 8-OHdG and 3-NT which efficiently enhanced the antioxidative defense system and protected the HSFs from subsequent oxidative stress damage. In other words, hydrogen decreased the excessive generation of intracellular O(2)(-) and elevated the cellular antioxidative defense. Based on our results, hydrogen may have applications in the treatment of skin diseases caused by diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Antioxidants / pharmacology*
  • Cell Proliferation*
  • Culture Media / pharmacology
  • DNA Damage / drug effects
  • Deoxyguanosine / analogs & derivatives
  • Deoxyguanosine / analysis
  • Diabetes Complications / pathology
  • Diabetes Complications / prevention & control
  • Fibroblasts / drug effects*
  • Glucose / pharmacology
  • Glutathione / analysis
  • Humans
  • Hydrogen / pharmacology*
  • Malondialdehyde / analysis
  • Mannitol / pharmacology
  • Oxidative Stress / drug effects*
  • Reactive Oxygen Species / metabolism
  • Skin / drug effects*
  • Skin Diseases / pathology
  • Skin Diseases / prevention & control
  • Superoxide Dismutase / analysis
  • Tyrosine / analogs & derivatives
  • Tyrosine / analysis


  • Antioxidants
  • Culture Media
  • Reactive Oxygen Species
  • 3-nitrotyrosine
  • Mannitol
  • Tyrosine
  • Malondialdehyde
  • Hydrogen
  • 8-Hydroxy-2'-Deoxyguanosine
  • Superoxide Dismutase
  • Deoxyguanosine
  • Glutathione
  • Glucose