Gene transfer of IkappaBalpha limits infarct size in a mouse model of myocardial ischemia-reperfusion injury

Lab Invest. 2003 Aug;83(8):1097-104. doi: 10.1097/01.lab.0000082060.39079.a6.

Abstract

Nuclear factor-kappaB (NF-kappaB) plays a central role in myocardial ischemia-reperfusion (MI/R) injury. The inhibitory protein IkappaBalpha prevents its activation. We investigated the effects of adeno-associated viral vector-mediated IkappaBalpha gene transfer in MI/R injury. Male C57BL/6 mice were randomized to receive a recombinant adeno-associated virus (rAAV) encoding the gene for the NF-kappaB inhibitory protein IkappaBalpha (rAAV- IkappaBalpha) or the beta-galactosidase gene (a control and inert gene; rAAV-LacZ), both at a dose of 10(11) copies. Four weeks later anesthetized animals were subjected to total occlusion (45 minutes) of the left main coronary artery followed by 5 hours of reperfusion. MI/R produced a wide infarct size (IF/area-at-risk = 56 +/- 8%; IF/left ventricle = 44 +/- 5%) and tissue neutrophil infiltration, studied by means of elastase activity (area-at-risk = 2.5 +/- 0.4 micro g/gm tissue; infarct area = 2.9 +/- 0.6 micro g/gm tissue). Furthermore MI/R caused peak message for intercellular adhesion molecule-1 (ICAM-1) in the area-at-risk at 3 hours of reperfusion (1.2 +/- 0.4 relative amount of cardiac ICAM-1 mRNA). NF-kappaB activation was evident at 0.5 hours of reperfusion and reached its maximum increase at 2 hours of reperfusion. rAAV-IkappaBalpha injection reduced infarct size (IF/area-at-risk = 19 +/- 3%; IF/left ventricle = 10 +/- 2%; p < 0.001), blocked NF-kappaB activation, diminished cardiac ICAM-1 expression (0.4 +/- 0.02 relative amount of cardiac ICAM-1 mRNA; p < 0.001), and blunted leukocyte accumulation (area-at-risk = 0.6 +/- 0.05 micro g/gm tissue; infarct area = 0.4 +/- 0.02 micro g/gm tissue; p < 0.001). Our data indicate that rAAV-IkappaBalpha may be useful for MI/R gene therapy.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Blotting, Western
  • DNA Primers / chemistry
  • Gene Transfer Techniques
  • Genetic Therapy*
  • Heart Ventricles / metabolism
  • Heart Ventricles / pathology
  • I-kappa B Proteins / genetics*
  • Intercellular Adhesion Molecule-1 / genetics
  • Intercellular Adhesion Molecule-1 / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Myocardial Infarction / genetics*
  • Myocardial Infarction / pathology
  • Myocardial Infarction / prevention & control*
  • Myocardial Reperfusion Injury*
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / metabolism
  • Pancreatic Elastase / metabolism
  • RNA, Messenger / metabolism
  • Transduction, Genetic / methods*
  • beta-Galactosidase / metabolism

Substances

  • DNA Primers
  • I-kappa B Proteins
  • NF-kappa B
  • Nfkbia protein, mouse
  • RNA, Messenger
  • Intercellular Adhesion Molecule-1
  • NF-KappaB Inhibitor alpha
  • beta-Galactosidase
  • Pancreatic Elastase