Hydrogen gas inhalation alleviates myocardial ischemia-reperfusion injury by the inhibition of oxidative stress and NLRP3-mediated pyroptosis in rats

Life Sci. 2021 May 1;272:119248. doi: 10.1016/j.lfs.2021.119248. Epub 2021 Feb 20.


Aims: Reperfusion therapy is the most common and effective treatment against ischemic heart disease (IHD), but the process inflicts massive ischemia/reperfusion (I/R) injury for which no treatment exists. Notably, reperfusion after ischemia causes ischemia/reperfusion injury (IR injury) and the "no-reflow" phenomenon seriously affecting the therapeutic effects in clinical practice. The principle purpose of this study is to validate the effect of hydrogen gas on IHD and further explore the mechanism of hydrogen gas in alleviating myocardial I/R injury and no-reflow phenomenon.

Materials and methods: The rat model of myocardial ischemia-reperfusion was well established. Myocardial infarct size was evaluated by TTC & Evans blue staining. The no-reflow area and the cardiac function were assessed by thioflavin-S staining and echocardiography respectively. Microstructure and mitochondria of myocardial tissue were assessed by transmission electron microscope. Western blot and immunohistochemistry were used to evaluate the expression of NLRP3 mediated pyroptosis related proteins. The 8-OHdG, MDA and serum total ROS were used to evaluate the degree of oxidative stress.

Key findings: The myocardial infarct size, no-reflow area, cardiac function, microstructure and mitochondrial morphology of I/R model rats were significantly improved after hydrogen inhalation. In addition, the expression of 8-OHdG, MDA, ROS and NLRP3 mediated pyroptosis related proteins were significantly decreased.

Significance: We found that oxidative stress and NLRP3 mediated pyroptosis are the important mechanisms for hydrogen to alleviate myocardial I/R injury, and we also confirmed that hydrogen can significantly improve no reflow phenomenon caused by ischemia-reperfusion.

Keywords: Hydrogen; Myocardial ischemia-reperfusion injury; NLRP3 inflammasome; Oxidative stress; Pyroptosis; The no-reflow phenomenon.

MeSH terms

  • Administration, Inhalation
  • Animals
  • Cell Survival / drug effects
  • Hydrogen / administration & dosage
  • Hydrogen / pharmacology*
  • Inflammasomes / metabolism
  • Male
  • Myocardial Infarction / metabolism
  • Myocardial Ischemia / drug therapy*
  • Myocardial Reperfusion Injury / metabolism
  • Myocytes, Cardiac / metabolism
  • NLR Family, Pyrin Domain-Containing 3 Protein / metabolism
  • Oxidative Stress / drug effects
  • Pyroptosis / drug effects
  • Rats
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species / metabolism
  • Reperfusion Injury / drug therapy
  • Reperfusion Injury / prevention & control


  • Inflammasomes
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Nlrp3 protein, rat
  • Reactive Oxygen Species
  • Hydrogen