In vivo reduction of the nuclear factor-kappaB activity using synthetic cis-element decoy oligonucleotides suppresses intimal hyperplasia in the injured carotid arteries in rabbits.
OBJECTIVE
Nuclear factor-kappaB (NF-kappaB) plays a critical role in inflammation-related reactions, and is also found in the injured arterial wall. The purpose of this study was to introduce synthetic double-stranded cis-element "decoy" oligonucleotides (ODNs) into the arterial wall using the hemagglutinating virus of Japan (HVJ) liposome, and to investigate the inhibitory potential of decoy ODN against balloon injury-induced intimal hyperplasia by reducing NF-kappaB activity.


METHODS
Fluorescein isothiocyanate (FITC)-labeled decoy ODNs using the HVJ-liposome method were tranfected in balloon-injured rabbit carotid arteries. We then performed electrophoretic mobility shift assay to examine NF-kappaB activity using balloon-injured arteries, and we introduced NF-kappaB decoy into balloon-injured arteries.


RESULTS
Transfection of FITC-labeled decoy ODNs by using the HVJ-liposome method demonstrated highly efficient protein expression with diffuse, frequent, and widespread nuclear signals over the entire medial layer, while the same amount of naked ODNs showed much less efficiency with scattered distribution of fluorescence in balloon-injured carotid arteries. Electrophoretic mobility shift assay showed activation of NF-kappaB in balloon-injured arteries. In vivo transfection of decoy ODNs mediated by HVJ liposome abolished the NF-kappaB activity in injured arteries with specific binding affinity to NF-kappaB protein. Intimal hyperplasia of carotid artery after balloon injury was reduced by approximately 50% by NF-kappaB decoy transfection compared with buffer treatment or scrambled decoy transfection.


CONCLUSIONS
Our results demonstrated involvement of NF-kappaB in intimal formation after arterial injury, and indicated that NF-kappaB can be an appropriate molecular target for anti-restenosis therapy.
Takeuchi
Kensuke
K
Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
Itoh
Hiroyuki
H
Yonemitsu
Yoshikazu
Y
Matsumoto
Takuya
T
Kume
Masazumi
M
Komori
Kimihiro
K
Maehara
Yoshihiko
Y
D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D017209:Apoptosis; D001665:Binding Sites / Q000235:genetics; D020212:Carotid Artery Injuries / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D017536:Carotid Artery, Common; D004195:Disease Models, Animal; D018450:Disease Progression; D004586:Electrophoresis; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006965:Hyperplasia; D020287:In Situ Nick-End Labeling; D007267:Injections; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage*; D011817:Rabbits; D017539:Tunica Intima / Q000187:drug effects / Q000473:pathology*
D005456:Fluorescent Dyes; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D016650:Fluorescein-5-isothiocyanate