Caspase-3-mediated splenic lymphocyte apoptosis in a porcine model of cardiac arrest

Am J Emerg Med. 2014 Sep;32(9):1027-32. doi: 10.1016/j.ajem.2014.06.004. Epub 2014 Jun 12.

Abstract

Background: Postresuscitation immunologic dysfunction contributes to the low survival rate after successful resuscitation, but its mechanism remains poorly understood. The mitochondrial apoptosis pathway is initiated by the Bcl-2/Bax-controlled and caspase-3-mediated pathway, this study investigated whether mitochondrial pathway-mediated splenic lymphocyte apoptosis is involved in the postresuscitation immunosuppression in a porcine model of cardiac arrest.

Methods: Twenty-eight Wuzhishan miniature pigs were randomly divided into 2 groups: return of spontaneous circulation (ROSC; n = 22) and sham-operated (n = 6). Return of spontaneous circulation was initiated after 8 minutes of untreated ventricular fibrillation. After successful ROSC, CD4(+) and CD8(+) lymphocyte subsets were determined by flow cytometry. Surviving pigs were randomly assigned to be humanely killed at 24 and 72 hours after ROSC (n = 8 per group). Spleens were removed for histopathologic analysis, Western blotting, quantitative real-time polymerase chain reaction, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay.

Results: A high degree of splenic lymphocyte apoptosis was observed in the ROSC group. Expression of Bax and activated caspase-3 was markedly increased in splenic tissue, whereas Bcl-2 was significantly decreased in the post-ROSC group compared with the sham-operated group (P < .05) at 24 and 72 hours after ROSC. The messenger RNA levels of activated caspase-3 of splenic tissue were significantly elevated at 24 and 72 hours after ROSC.

Conclusion: These results demonstrates that Bcl-2/Bax and caspase-3-mediated mitochondrial apoptosis signaling pathway may contribute to abnormal splenic lymphocyte apoptosis, which may be one of the main pathologic mechanisms of postresuscitation disturbance of immunologic function in a porcine model of cardiac arrest.

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Blotting, Western
  • CD4 Lymphocyte Count
  • CD4-Positive T-Lymphocytes / physiology
  • CD8-Positive T-Lymphocytes / physiology
  • Caspase 3 / physiology*
  • Disease Models, Animal
  • Flow Cytometry
  • Heart Arrest / immunology
  • Heart Arrest / physiopathology*
  • Lymphocytes / physiology*
  • Male
  • Proto-Oncogene Proteins c-bcl-2 / physiology
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction / physiology
  • Spleen / cytology
  • Spleen / physiopathology*
  • Swine
  • Swine, Miniature
  • bcl-2-Associated X Protein / physiology

Substances

  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Caspase 3