A novel strategy for myocardial protection using in vivo transfection of cis element 'decoy' against NFkappaB binding site: evidence for a role of NFkappaB in ischemia-reperfusion injury.
BACKGROUND
NFkappaB, an important transcriptional factor, has been reported to play a significant role in the coordinated transcription of cytokine and adhesion molecule genes. Therefore, blocking the NFkappaB may attenuate ischemia reperfusion injury in the myocardium. For blocking transcriptional factors, gene therapy, such as cis element "decoy," appears to be an innovative and useful therapy. This study aimed to prove the efficacy of cis element decoy against NFkappaB binding site for myocardial protection.


RESULTS
Rat hearts were transfected with fluorescence isothiocyanate-labeled cis element decoy against NFkappaB (NF)-binding site (NF group, n=6) and scrambled decoy (SD) group (n=6) by coronary infusion of hemagglutinating virus of Japan (HVJ)-liposome during cardioplegic arrest. Both the NF and SD groups showed marked FITC-staining in the nuclei of myocytes, demonstrating the efficacy of gene transfer into the nuclei of cardiac myocytes as compared with the control group transfected with empty liposomes. After 3 days of transfection, the NF group showed significantly higher percentages of recovery of left ventricular developed pressure (NF versus SD, 87+/-11 versus 54+/-12%) and coronary flow (97+/-16 versus 61+/-15%) than did the control hearts when exposed to ischemia (30 minutes, 37 degrees C) and reperfusion (30 minutes, 37 degrees C). The NF group showed a significantly lower percentage of neutrophil adherence to endothelial cells (38+/-6 versus 81+/-3%) and a lower tissue level of interleukin-8 (109+/-48 versus 210+/-55 ng/mg) than did the SD group.


CONCLUSIONS
The hearts transfected with cis element decoy against NFkappaB binding site showed significant improvement in tolerance against ischemia-reperfusion injury in association with the inhibition of neutrophil adherence and tissue IL-8 production. This suggests that NFkappaB plays a significant role in ischemia-reperfusion injury. This method, using in vivo gene transfection of cis element decoy against NFkappaB binding site, appears to be a novel and future strategy for myocardial protection.
Sawa
Y
Y
First Department of Surgery, Institute for Cellular and Molecular Biology, and Fujisawa Pharmaceutical Co, Ltd, Osaka University Medical School, Suita, Japan.
Morishita
R
R
Suzuki
K
K
Kagisaki
K
K
Kaneda
Y
Y
Maeda
K
K
Kadoba
K
K
Matsuda
H
H
D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D015316:Genetic Therapy; D016209:Interleukin-8 / Q000096:biosynthesis; D015428:Myocardial Reperfusion Injury / Q000209:etiology*; D016328:NF-kappa B / Q000235:genetics / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology; D009841:Oligonucleotides / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection
D016209:Interleukin-8; D016328:NF-kappa B; D009841:Oligonucleotides