Ephrin type-B receptor 4 activation reduces neointimal hyperplasia in human saphenous vein in vitro.
Ephrin type-B receptor 4 activation reduces neointimal hyperplasia in human saphenous vein in vitro.
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DOI：
10.1016/j.jvs.2014.09.036
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发表时间：
2016-03
影响因子：
4.3
通讯作者：
Dardik, Alan
中科院分区：
文献类型：
--
作者：
Wong, Daniel J.;Lu, Daniel Y.;Protack, Clinton D.;Kuwahara, Go;Bai, Hualong;Sadaghianloo, Nirvana;Tellides, George;Dardik, Alan
尽管出现新生内膜增生，静脉搭桥术仍然是晚期外周动脉和冠状动脉疾病患者的基本治疗方法。我们已经证明，用其配体 Ephrin-B2 刺激受体酪氨酸激酶 Eph-B4 可防止小鼠静脉移植物中的新内膜增生。本研究确定人静脉中的 Eph-B4 是否能够磷酸化、激活下游信号通路，以及在体外释放一氧化氮和预防新生内膜增生的功能。从手术室取出废弃的人隐静脉，置于不含或含 Ephrin-B2/Fc（2 μg/ml；14 天）的器官培养物中，并测量匹配静脉中的新内膜：中膜比率。用 Ephrin-B2/Fc (2 μg/ml) 处理原代人脐静脉内皮细胞 (HUVEC)，并通过 qPCR、蛋白质印迹、免疫分析和一氧化氮释放进行检查。将 Ephrin-B2/Fc (2 μg/ml) 置于隐静脉外膜上的 Pluronic 凝胶中，在生物反应器中用动脉剪切应力处理 24 小时，并用免疫荧光检查激活的 Eph-B4。隐静脉环的基线内膜：中膜比率为 0.456 ± 0.097，在器官培养 14 天后，未处理的静脉中增加到 0.726 ± 0.142，但在用 Ephrin-B2/Fc 处理的静脉中仅增加到 0.630 ± 0.132 (p=.017; n=19)。 Ephrin-B2/Fc 刺激 Akt、eNOS 和 Caveolin-1 磷酸化以及 HUVEC (n=6) 的 NO 释放 (p=0.007)。在生物反应器中动脉应激 24 小时后，递送至外膜的 Ephrin-B2/Fc 刺激内皮 Eph-B4 磷酸化 (n=3)。 Eph-B4 存在于成人隐静脉中并具有功能，具有完整的下游信号通路，能够在体外释放一氧化氮并预防新内膜增生。 Ephrin-B2/Fc 的外膜递送可激活体外经动脉剪切应力处理的隐静脉中的内皮 Eph-B4。这些结果表明，刺激 Eph-B4 功能可能是转化为旨在抑制静脉新内膜增生的人体临床试验的候选策略。
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 Eph-B4 with its ligand Ephrin-B2 prevents neointimal hyperplasia in murine vein grafts. This study determines whether Eph-B4 in human veins is capable of phosphorylation, activation of downstream signaling pathways, and functional to release nitric oxide and prevent neointimal hyperplasia in vitro. Discarded human saphenous veins were taken from the operating room and placed in organ culture without or with Ephrin-B2/Fc (2 μg/ml; 14 days) and the neointima:media ratio was measured in matched veins. Primary human umbilical vein endothelial cells (HUVEC) were treated with Ephrin-B2/Fc (2 μg/ml) and examined with qPCR, Western blot, immunoassays and for release of nitric oxide. Ephrin-B2/Fc (2 μg/ml) was placed in pluronic gel on the adventitia of saphenous veins treated with arterial shear stress for 24 hours in a bioreactor and activated Eph-B4 examined with immunofluorescence. 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 (p=.017; n=19). Ephrin-B2/Fc stimulated Akt, eNOS and caveolin-1 phosphorylation and NO release (p=0.007) from HUVEC (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). Eph-B4 is present and functional in adult human saphenous veins, with intact downstream signaling pathways capable of nitric oxide 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.
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影响因子：
2.2
作者：
Li, Fan Dong;Sexton, Kevin W.;Hocking, Kyle M.;Osgood, Michael J.;Eagle, Susan;Cheung-Flynn, Joyce;Brophy, Colleen M.;Komalavilas, Padmini
通讯作者：
Komalavilas, Padmini
影响因子：
16
作者：
Gerety, SS;Wang, HU;Anderson, DJ
通讯作者：
Anderson, DJ
影响因子：
64.8
作者：
Dimmeler, S;Fleming, I;Zeiher, AM
通讯作者：
Zeiher, AM
DOI：
10.1016/0735-1097(96)00206-9
发表时间：
1996-09-01
影响因子：
24
作者：
FitzGibbon, GM;Kafka, HP;Burton, JR
通讯作者：
Burton, JR
影响因子：
4.8
作者：
Fulton, D;Babbitt, R;Sessa, WC
通讯作者：
Sessa, WC