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Ephrin type-B receptor 4 activation reduces neointimal hyperplasia in human saphenous vein in vitro.
- Source :
-
Journal of vascular surgery [J Vasc Surg] 2016 Mar; Vol. 63 (3), pp. 795-804. Date of Electronic Publication: 2014 Oct 24. - Publication Year :
- 2016
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Abstract
- Background: Vein bypass is an essential therapy for patients with advanced peripheral and coronary artery disease despite development of neointimal hyperplasia. We have shown that stimulation of the receptor tyrosine kinase ephrin type-B receptor 4 (Eph-B4) with its ligand ephrin-B2 prevents neointimal hyperplasia in murine vein grafts. This study determines whether Eph-B4 in adult human veins is capable of phosphorylation and activation of downstream signaling pathways, as well as functional to release nitric oxide (NO) and prevent neointimal hyperplasia in vitro.<br />Methods: Discarded human saphenous veins were taken from the operating room and placed in organ culture without or with ephrin-B2/Fc (2 μg/mL) for 14 days, and the neointima/media ratio was measured in matched veins. Primary human umbilical vein endothelial cells were treated with ephrin-B2/Fc (2 μg/mL) and examined with quantitative polymerase chain reaction, Western blot, immunoassays, and for release of NO. Ephrin-B2/Fc (2 μg/mL) was placed on the adventitia of saphenous veins treated with arterial shear stress for 24 hours in a bioreactor and activated Eph-B4 examined with immunofluorescence.<br />Results: The baseline intima/media ratio in saphenous vein rings was 0.456 ± 0.097, which increased to 0.726 ± 0.142 in untreated veins after 14 days in organ culture but only to 0.630 ± 0.132 in veins treated with ephrin-B2/Fc (n = 19, P = .017). Ephrin-B2/Fc stimulated Akt, endothelial NO synthase and caveolin-1 phosphorylation, and NO release (P = .007) from human umbilical vein endothelial cells (n = 6). Ephrin-B2/Fc delivered to the adventitia stimulated endothelial Eph-B4 phosphorylation after 24 hours of arterial stress in a bioreactor (n = 3).<br />Conclusions: Eph-B4 is present and functional in adult human saphenous veins, with intact downstream signaling pathways capable of NO release and prevention of neointimal hyperplasia in vitro. Adventitial delivery of ephrin-B2/Fc activates endothelial Eph-B4 in saphenous veins treated with arterial shear stress in vitro. These results suggest that stimulation of Eph-B4 function may be a candidate strategy for translation to human clinical trials designed to inhibit venous neointimal hyperplasia.<br /> (Published by Elsevier Inc.)
- Subjects :
- Bioreactors
Caveolin 1 metabolism
Cell Proliferation drug effects
Cells, Cultured
Enzyme Activation
Human Umbilical Vein Endothelial Cells drug effects
Human Umbilical Vein Endothelial Cells pathology
Humans
Hyperplasia
Mechanotransduction, Cellular drug effects
Nitric Oxide metabolism
Nitric Oxide Synthase Type III metabolism
Phosphorylation
Primary Cell Culture
Proto-Oncogene Proteins c-akt metabolism
Receptor, EphB4 genetics
Receptor, EphB4 metabolism
Saphenous Vein metabolism
Saphenous Vein pathology
Stress, Mechanical
Tissue Culture Techniques instrumentation
Ephrin-B2 pharmacology
Immunoglobulin Fc Fragments pharmacology
Neointima
Receptor, EphB4 agonists
Saphenous Vein drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1097-6809
- Volume :
- 63
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Journal of vascular surgery
- Publication Type :
- Academic Journal
- Accession number :
- 25446283
- Full Text :
- https://doi.org/10.1016/j.jvs.2014.09.036