基质金属蛋白酶-8 促进血管平滑肌细胞增殖和新生内膜形成。
Matrix metalloproteinase-8 promotes vascular smooth muscle cell proliferation and neointima formation.
机构信息
From William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom (Q. Xiao, F.Z., S.Y.); Department of Cardiology, Peking University People's Hospital, Beijing, China (F.Z.); Department of Pharmacy, University of Naples Federico II, Naples, Italy (G.G., M. Maddaluno, P.M., A.I.); Cardiovascular Division, King's College London BHF Centre, London, United Kingdom (Y.H., Z.Z., Q. Xing, X.Y., M. Mayr, Q. Xu); Clemens Schöpf Institute, Technische Universität Darmstadt, Darmstadt, Germany (B.D., B.S.); Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom (P.M.).
出版信息
Arterioscler Thromb Vasc Biol. 2014 Jan;34(1):90-8. doi: 10.1161/ATVBAHA.113.301418. Epub 2013 Oct 24.
OBJECTIVE
We investigated the role of matrix metalloproteinase-8 (MMP8) in neointima formation and in vascular smooth muscle cell (VSMC) migration and proliferation.
APPROACH AND RESULTS
After carotid artery wire injuring, MMP8(-/-)/apoE(-/-) mice had fewer proliferating cells in neointimal lesions and smaller lesion sizes. Ex vivo assays comparing VSMCs isolated from MMP8 knockout and wild-type mice showed that MMP8 knockout decreased proliferation and migration. Proteomics analysis revealed that a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) had lower concentrations in MMP8 knockout VSMC culture media than in MMP8 wild-type VSMC culture media. Western blot, flow cytometric, and immunocytochemical analyses showed that MMP8 knockout VSMCs contained more pro-ADAM10 but less mature ADAM10, more N-cadherin, and β-catenin in the plasma membrane but less β-catenin in the nucleus and less cyclin D1. Treatment of MMP8 wild-type VSMCs with an ADAM10 inhibitor, GI254023X, or siRNA knockdown of ADAM10 in MMP8 wild-type VSMCs inhibited proliferation and migration, increased N-cadherin and β-catenin in the plasma membrane, reduced β-catenin in the nucleus, and decreased cyclin D1 expression. Incubation of MMP8 knockout VSMCs with a recombinant ADAM10 rescued the proliferative and migratory ability of MMP8 knockout VSMCs and increased cyclin D1 expression. Furthermore, immunohistochemical analyses showed colocalization of ADAM10 with VSMCs and N-cadherin, and nuclear accumulation of β-catenin in the neointima in apoE(-/-)/MMP8(+/+) mice.
CONCLUSIONS
MMP8 enhances VSMC proliferation via an ADAM10, N-cadherin, and β-catenin-mediated pathway and plays an important role in neointima formation.
目的
研究基质金属蛋白酶 8（MMP8）在新生内膜形成以及血管平滑肌细胞（VSMC）迁移和增殖中的作用。
方法和结果
颈总动脉线损伤后，MMP8（-/-）/apoE（-/-）小鼠的新生内膜病变中增殖细胞较少，病变较小。比较 MMP8 敲除和野生型小鼠分离的 VSMC 的离体实验表明，MMP8 敲除降低了增殖和迁移。蛋白质组学分析显示，MMP8 敲除 VSMC 培养基中的解整合素金属蛋白酶域蛋白 10（ADAM10）浓度低于 MMP8 野生型 VSMC 培养基。Western blot、流式细胞术和免疫细胞化学分析表明，MMP8 敲除 VSMC 含有更多的 pro-ADAM10 但成熟的 ADAM10 较少，质膜中 N-钙黏蛋白和β-连环蛋白较多，但核内β-连环蛋白较少，cyclin D1 较少。用 ADAM10 抑制剂 GI254023X 处理 MMP8 野生型 VSMC，或用 siRNA 敲低 MMP8 野生型 VSMC 中的 ADAM10，均可抑制增殖和迁移，增加质膜中的 N-钙黏蛋白和β-连环蛋白，减少核内β-连环蛋白，降低 cyclin D1 表达。用重组 ADAM10 孵育 MMP8 敲除 VSMC，可恢复 MMP8 敲除 VSMC 的增殖和迁移能力，并增加 cyclin D1 表达。此外，免疫组织化学分析显示 ADAM10 与 VSMC 和 N-钙黏蛋白的共定位，以及 apoE（-/-）/MMP8（+/+）小鼠新生内膜中β-连环蛋白的核内积聚。
结论
MMP8 通过 ADAM10、N-钙黏蛋白和β-连环蛋白介导的途径增强 VSMC 增殖，并在新生内膜形成中发挥重要作用。
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