Transfection of nuclear factor-kappaB decoy oligodeoxynucleotide protects against ischemia/reperfusion injury in a rat epigastric flap model

J Gene Med. 2012 Nov;14(11):623-31. doi: 10.1002/jgm.2677.

Abstract

Background: Nuclear factor-kappaB (NF-κB) is considered to play an important role in the response to ischemia/reperfusion (I/R) injury in flap surgery. To inhibit NF-κB, synthetic double-stranded oligodeoxynucleotide (ODN) was used as a decoy. The present study aimed to evaluate the suppressive effects of NF-κB against I/R injury of experimental rat flap model.

Methods: An extended epigastric island flap was raised and ischemia was induced for 3 h. NF-κB decoy ODN (group D) or single-strand ODN (control; group S) was injected via the contralateral artery when the pedicle was clamped. Transfection efficiency was evaluated with fluorescein isothiocyanate (FITC)-labeled ODN. The effects of NF-κB decoy ODN were analyzed in groups D and S, and an untreated group (group N).

Results: FITC-labeled ODN was distributed over the entire flap. Mean survival rate of the flap was significantly higher in group D than in the other groups (group D: 57.9%; group S: 31.1%; group N 31.7%; p < 0.005). Injured muscle fibers, neutrophils and the expression of inducible nitric oxide synthase were significantly lower in group D. A real-time polymerase chain reaction also demonstrated a tendency for suppression of tumor necrosis factor-α, interleukin (IL)-1β and IL-6.

Conclusions: We show that NF-κB decoy ODN protected against flap necrosis as a result of I/R injury in rats. We also indicate that intra-arterial injection of naked NF-κB decoy ODN is effective for transfection into target organs. Therefore, transfection of NF-κB decoy ODN represents a novel therapeutic strategy for the treatment of flap surgery in I/R injury.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Genetic Therapy / methods*
  • Inflammation / metabolism
  • Inflammation / prevention & control
  • Male
  • NF-kappa B / antagonists & inhibitors*
  • Neutrophils / drug effects
  • Neutrophils / metabolism
  • Nitric Oxide Synthase Type II / metabolism
  • Oligodeoxyribonucleotides / biosynthesis
  • Oligodeoxyribonucleotides / genetics*
  • Oxidative Stress / drug effects
  • Rats
  • Rats, Sprague-Dawley
  • Reperfusion Injury / genetics
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / therapy*
  • Surgical Flaps
  • Transfection / methods

Substances

  • NF-kappa B
  • NF-kappaB decoy
  • Oligodeoxyribonucleotides
  • Nitric Oxide Synthase Type II
  • Nos2 protein, rat