Ex vivo blockade of endothelin-1 inhibits graft coronary artery disease in a rodent cardiac allograft model.
BACKGROUND
Graft coronary artery disease (GCAD) is characterized by vascular narrowing resulting from intimal hyperplasia. Endothelin (ET)-1, derived from the vascular endothelium and macrophages, stimulates vascular smooth muscle cells (SMCs), which leads to neointimal formation in donor graft coronary arteries. In this study, we hypothesized that antisense (AS) oligodeoxynucleotides (ODN) for preproendothelin-1 (ppET-1) delivered to rat cardiac allografts by means of hyperbaric pressure would reduce the incidence of GCAD.


METHODS
PVG donor hearts were infused with ppET-1 AS ODN (80 micromol/liter), sense ODN, scrambled ODN or saline alone and incubated in a pressure chamber at 75 psi or ambient pressure for 45 minutes. Cardiac allografts were heterotopically transplanted into ACI rats treated with cyclosporine (7.5 mg/kg, Days 0 to 9). Allografts were procured at post-operative days (POD) 7 or 90. Reverse transcription-polymerase chain reaction (RT-PCR) assay for ET-1 mRNA and ET01 immunohistochemistry (IHC) were performed at PODs 7 and 90. Elastic staining and IHC with anti-macrophage and alpha-SMC actin antibodies were performed to assess GCAD at POD 90.


RESULTS
Treatment with AS ODN and pressure significantly reduced ET-1 mRNA and protein expression. A significant reduction in GCAD was achieved with inhibition of ET-1 and was associated with attenuation of macrophages and SMCs in the neointima.


CONCLUSIONS
Peri-operative ex vivo inhibition of ET-1 expression results in a reduction of GCAD. This highly targeted therapy may be a clinically viable strategy for the prevention of ET-1-induced GCAD following cardiac transplantation.
Yamaguchi
Atsushi
A
Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, California, USA.
Miniati
Douglas N
DN
Hirata
Ken ichi
Ki
Hoyt
E Grant
EG
Robbins
Robert C
RC
D000818:Animals; D003324:Coronary Artery Disease / Q000209:etiology* / Q000628:therapy*; D004195:Disease Models, Animal; D019332:Endothelin-1 / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000627:therapeutic use*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D016027:Heart Transplantation; D007150:Immunohistochemistry; D008297:Male; D008955:Models, Cardiovascular; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D011183:Postoperative Complications / Q000209:etiology / Q000378:metabolism / Q000628:therapy; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
D019332:Endothelin-1; D012333:RNA, Messenger