Bock Felix, Onderka Jasmine, Dietrich Tina, Bachmann Björn, Kruse Friedrich E, Paschke Matthias, Zahn Grit, Cursiefen Claus
Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany.
Invest Ophthalmol Vis Sci. 2007 Jun;48(6):2545-52. doi: 10.1167/iovs.06-0570.
To analyze whether bevacizumab can inhibit inflammatory angiogenesis and lymphangiogenesis in the cornea. Bevacizumab (Avastin; Roche, Welwyn Garden City, UK) is a recombinant, humanized, monoclonal antibody against VEGF-A that has been approved by the U.S. Food and Drug Administration for the treatment of colon carcinomas.
The mouse model of suture-induced corneal neovascularization was used to assess the antihemangiogenic and antilymphangiogenic effect of bevacizumab by systemic and topical application. Corneal flatmounts were stained with LYVE-1 as a specific lymphatic vascular endothelial marker and CD31 as a pan-endothelial marker, and blood and lymph vascularized areas were analyzed morphometrically. The inhibitory effect of bevacizumab on lymphatic endothelial cells (LECs) was analyzed with a colorimetric (BrdU) proliferation ELISA. The binding ability of bevacizumab to murine VEGF-A was analyzed by Western blot, ELISA, and surface plasmon resonance.
The systemic and topical applications of bevacizumab significantly inhibited the outgrowth of blood (P < 0.006 and P < 0.0001, respectively) and lymphatic (P < 0.002 and P < 0.0001, respectively) vessels. Inhibition of the proliferation of LECs was also significant (P < 0.0001). Western blot analysis, ELISA, and the surface plasmon resonance assay showed that bevacizumab binds murine VEGF-A.
Topical or systemic application of bevacizumab inhibits both inflammation-induced angiogenesis and lymphangiogenesis in the cornea. This finding suggests an important role of VEGF-A in corneal lymphangiogenesis. Bevacizumab may be useful in preventing immune rejections after penetrating keratoplasty or tumor metastasis via lymphatic vessels.
分析贝伐单抗是否能抑制角膜中的炎性血管生成和淋巴管生成。贝伐单抗（阿瓦斯汀；罗氏公司，英国韦林花园城）是一种重组人源化抗VEGF-A单克隆抗体，已被美国食品药品监督管理局批准用于治疗结肠癌。
采用缝线诱导角膜新生血管小鼠模型，通过全身和局部应用来评估贝伐单抗的抗血管生成和抗淋巴管生成作用。角膜铺片用LYVE-1（一种特异性淋巴管内皮标志物）和CD31（一种泛内皮标志物）染色，对血管化和淋巴管化区域进行形态计量分析。用比色法（BrdU）增殖ELISA分析贝伐单抗对淋巴管内皮细胞（LEC）的抑制作用。通过蛋白质印迹法、ELISA和表面等离子体共振分析贝伐单抗与小鼠VEGF-A的结合能力。
贝伐单抗的全身和局部应用均显著抑制血管（分别为P < 0.006和P < 0.0001）和淋巴管（分别为P < 0.002和P < 0.0001）的生长。对LEC增殖的抑制也很显著（P < 0.0001）。蛋白质印迹分析、ELISA和表面等离子体共振分析表明贝伐单抗能结合小鼠VEGF-A。
贝伐单抗局部或全身应用均可抑制角膜中炎症诱导的血管生成和淋巴管生成。这一发现提示VEGF-A在角膜淋巴管生成中起重要作用。贝伐单抗可能有助于预防穿透性角膜移植术后的免疫排斥反应或肿瘤经淋巴管转移。
