Hydrogen Treatment Protects Mice Against Chronic Pancreatitis by Restoring Regulatory T Cells Loss

Cell Physiol Biochem. 2017;44(5):2005-2016. doi: 10.1159/000485906. Epub 2017 Dec 11.


Background/aims: Chronic pancreatitis is an inflammatory disease of the pancreas characterized by progressive tissue destruction and fibrogenesis. The development of chronic pancreatitis is associated with immune cell dysregulation. Currently, the specific and effective treatment of chronic pancreatitis remains absent.

Methods: By using an L-arginine induced chronic pancreatitis mouse model, we tested the therapeutic potential of hydrogen, a strong hydroxyl radicals scavenger, in the chronic pancreatitis model. Tissue inflammation, damage and fibrosis were analyzed on HE, TUNEL, MPO, and sirius staining. Pancreas levels of MDA content, SOD activity, TNF-α , IL-10 cytokine expression and serum amylase and lipase activity were determined by ELISA and absorbance assay. Apoptosis, T cells subtype proportion and intracellular level of reactive oxygen species (ROS) were analyzed by flow cytometry. Tregs adoptive transfer and CD25 neutralization were used to validate the role of Tregs in chronic pancreatitis.

Results: We found that hydrogen treatment significantly improved multiple symptoms of chronic pancreatitis. The number of Tregs was reduced in chronic pancreatitis mice, while hydrogen treatment restored the Treg loss by L-arginine administrations. Depletion of Tregs abolished the protective effect of hydrogen treatment in chronic pancreatitis. In vitro study showed that hydrogen blocked ROS generation in Tregs and promoted Tregs survival.

Conclusion: Hydrogen treatment showed reliable benefits in controlling the severity of chronic pancreatitis. Our study supported that hydrogen could be used as a novel treatment in chronic pancreatitis patient in the future.

Keywords: Hydrogen; Immunoregulation; Pancreatitis; Treg.

MeSH terms

  • Adoptive Transfer
  • Amylases / blood
  • Animals
  • Antibodies, Neutralizing / immunology
  • Arginine / toxicity
  • Cell Survival / drug effects
  • Disease Models, Animal
  • Hydrogen / pharmacology*
  • Hydrogen / therapeutic use
  • Interleukin-10 / metabolism
  • Interleukin-2 Receptor alpha Subunit / immunology
  • Interleukin-2 Receptor alpha Subunit / metabolism
  • Lipase / blood
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Oxidative Stress / drug effects
  • Pancreas / metabolism
  • Pancreas / pathology
  • Pancreatitis, Chronic / chemically induced
  • Pancreatitis, Chronic / pathology
  • Pancreatitis, Chronic / prevention & control
  • Protective Agents / pharmacology*
  • Protective Agents / therapeutic use
  • Reactive Oxygen Species / metabolism
  • T-Lymphocytes, Regulatory / cytology
  • T-Lymphocytes, Regulatory / drug effects*
  • T-Lymphocytes, Regulatory / metabolism
  • Tumor Necrosis Factor-alpha / metabolism


  • Antibodies, Neutralizing
  • Interleukin-2 Receptor alpha Subunit
  • Protective Agents
  • Reactive Oxygen Species
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • Hydrogen
  • Arginine
  • Lipase
  • Amylases