Kong Jin-Sun, Yoo Seung-Ah, Kim Jung-Wook, Yang Seung-Pil, Chae Chi-Bom, Tarallo Valeria, De Falco Sandro, Ryu Sung-Ho, Cho Chul-Soo, Kim Wan-Uk
The Catholic University of Korea, Seoul, South Korea.
Arthritis Rheum. 2010 Jan;62(1):179-90. doi: 10.1002/art.27243.
To delineate the role of neuropilin-1 (NP-1), a vascular endothelial growth factor receptor (VEGFR), in rheumatoid inflammation and to determine whether blockade of NP-1 could suppress synoviocyte survival and angiogenesis.
VEGF(111-165) peptide, which encompasses the NP-1 binding domain of VEGF(165), was generated by cleaving VEGF(165) with plasmin. The effect of this peptide on the interaction between VEGF(165) and its receptor was determined by (125)I-VEGFR binding assay. Assays to determine synoviocyte apoptosis, adhesion, and migration were performed in the presence of VEGF(165) and/or the peptide. VEGF(165)-induced angiogenesis was assessed by measuring the proliferation, tube formation, and wounding migration of endothelial cells (ECs). Mice were immunized with type II collagen to induce experimental arthritis.
VEGF(111-165) peptide specifically inhibited the binding of (125)I-VEGF(165) to NP-1 on rheumatoid synoviocytes and ECs. The peptide eliminated the VEGF(165)-mediated increase in synoviocyte survival and activation of p-ERK and Bcl-2. The peptide also completely inhibited a VEGF(165)-induced increase in synoviocyte adhesion and migration. In addition, the anti-NP-1 peptide blocked VEGF(165)-stimulated proliferation, capillary tube formation, and wounding migration of ECs in vitro. VEGF(165)-induced neovascularization in a Matrigel plug in mice was also blocked by treatment with the peptide. Finally, subcutaneous injection of anti-NP-1 peptide suppressed arthritis severity and autoantibody formation in mice with experimental arthritis and inhibited synoviocyte hyperplasia and angiogenesis in arthritic joints.
Anti-NP-1 peptide suppressed VEGF(165)-induced increases in synoviocyte survival and angiogenesis, and thereby blocked experimental arthritis. Our findings suggest that anti-NP-1 peptide could be useful in alleviating chronic arthritis.
阐明血管内皮生长因子受体神经纤毛蛋白-1（NP-1）在类风湿性炎症中的作用，并确定阻断NP-1是否能抑制滑膜细胞存活和血管生成。
通过纤溶酶切割VEGF(165)产生包含VEGF(165)的NP-1结合域的VEGF(111 - 165)肽。通过(125)I-VEGFR结合试验确定该肽对VEGF(165)与其受体之间相互作用的影响。在VEGF(165)和/或该肽存在的情况下进行确定滑膜细胞凋亡、黏附及迁移的试验。通过测量内皮细胞（ECs）的增殖、管腔形成及创伤迁移来评估VEGF(165)诱导的血管生成。用II型胶原免疫小鼠以诱导实验性关节炎。
VEGF(111 - 165)肽特异性抑制(125)I-VEGF(165)与类风湿滑膜细胞和ECs上的NP-1结合。该肽消除了VEGF(165)介导的滑膜细胞存活增加以及p-ERK和Bcl-2的激活。该肽还完全抑制了VEGF(165)诱导的滑膜细胞黏附和迁移增加。此外，抗NP-1肽在体外阻断了VEGF(165)刺激的ECs增殖、毛细血管管腔形成及创伤迁移。用该肽处理也阻断了VEGF(165)在小鼠基质胶塞中诱导的新生血管形成。最后，皮下注射抗NP-1肽可抑制实验性关节炎小鼠的关节炎严重程度和自身抗体形成，并抑制关节炎关节中的滑膜细胞增生和血管生成。
抗NP-1肽抑制VEGF(165)诱导的滑膜细胞存活和血管生成增加，从而阻断实验性关节炎。我们的研究结果表明抗NP-1肽可能有助于缓解慢性关节炎。
