Bristol Heart Institute, University of Bristol, Bristol Royal Infirmary, Bristol, United Kingdom.
J Vasc Surg. 2010 Nov;52(5):1301-9. doi: 10.1016/j.jvs.2010.05.096. Epub 2010 Jul 13.
Inhibition of vascular smooth muscle cell (VSMC) migration is a potential strategy for reducing intimal thickening during in-stent restenosis and vein graft failure. In this study, we examined the effect of disrupting the function of the VSMC adhesion molecule, N-cadherin, using antagonists, neutralizing antibodies, and a dominant negative, on VSMC migration and intimal thickening. Migration was assessed by the scratch-wound assay of human saphenous vein VSMCs and in a human saphenous vein ex vivo organ culture model of intimal thickening.
Inhibition of cadherin function using a pan-cadherin antagonist, significantly reduced migration by 53%±8% compared with the control peptide (n=3; P<.05). Furthermore, inhibition of N-cadherin function with an N-cadherin antagonist, neutralizing antibodies, and adenoviral expression of dominant negative N-cadherin (RAd dn-N-cadherin), significantly reduced migration by 31%±2%, 23%±1% and 32%±7% compared with controls, respectively (n=3; P<.05). Inhibition of cadherin function significantly increased apoptosis by between 1.5- and 3.3-fold at the wound edge. In an ex vivo model of intimal thickening, inhibition of N-cadherin function by infection of human saphenous vein segments with RAd dn-N-cadherin significantly reduced VSMC migration by 55% and increased VSMC apoptosis by 2.7-fold. As a result, intimal thickening was significantly suppressed by 54%±14%. Importantly, there was no detrimental effect of dn-N-cadherin on endothelial coverage; in fact, it was significantly increased, as was survival of cultured human saphenous vein endothelial cells.
Under the condition of this study, cell-cell adhesion mediated by N-cadherin regulates VSMC migration via modulation of viability. Interestingly, inhibition of N-cadherin function significantly retards intimal thickening via inhibition of VSMC migration and promotion of endothelial cell survival. We suggest that disruption of N-cadherin-mediated cell-cell contacts is a potential strategy for reducing VSMC migration and intimal thickening.
抑制血管平滑肌细胞（VSMC）迁移是减少支架内再狭窄和静脉移植物失败时内膜增厚的一种潜在策略。在这项研究中，我们使用拮抗剂、中和抗体和显性负突变体来检测破坏 VSMC 黏附分子 N-钙黏蛋白功能对 VSMC 迁移和内膜增厚的影响。通过人隐静脉 VSMC 的划痕实验和人隐静脉内膜增厚的体外器官培养模型来评估迁移。
使用泛钙黏蛋白拮抗剂抑制钙黏蛋白功能，与对照肽相比，迁移减少了 53%±8%（n=3；P<.05）。此外，用 N-钙黏蛋白拮抗剂、中和抗体和腺病毒表达显性负突变体 N-钙黏蛋白（RAd dn-N-cadherin）抑制 N-钙黏蛋白功能，分别使迁移减少了 31%±2%、23%±1%和 32%±7%（n=3；P<.05）。钙黏蛋白功能的抑制使伤口边缘的细胞凋亡增加了 1.5-3.3 倍。在静脉内膜增厚的体外模型中，用 RAd dn-N-cadherin 感染人隐静脉段抑制 N-钙黏蛋白功能，使 VSMC 迁移减少 55%，VSMC 凋亡增加 2.7 倍，结果内膜增厚减少了 54%±14%。重要的是，dn-N-cadherin 对内皮细胞覆盖没有不良影响；事实上，它显著增加，培养的人隐静脉内皮细胞的存活率也显著增加。
在本研究条件下，N-钙黏蛋白介导的细胞-细胞黏附通过调节细胞活力来调节 VSMC 迁移。有趣的是，抑制 N-钙黏蛋白功能通过抑制 VSMC 迁移和促进内皮细胞存活，显著减缓内膜增厚。我们认为，破坏 N-钙黏蛋白介导的细胞-细胞接触是减少 VSMC 迁移和内膜增厚的一种潜在策略。
