Development of inhibitors for a angiogenesis factor CTGF and its application for angiogenetic diseases

1)作为结缔组织生长因子（CTGF）的抑制剂，合成CTGF片段免疫家兔制备多克隆抗体。此外，还制备了抗ctgf单克隆抗体。2)重组CTGF （rCTGF）促进血管内皮细胞的粘附、增殖和迁移，这些作用被抗CTGF抗体抑制。3) rCTGF显著诱导血管内皮细胞成管，且这种作用强于血管内皮生长因子对碱性成纤维细胞生长的作用。4)将rCTGF作用于鸡颈尿囊膜（CAM），可引起大体血管生成反应，且该作用被抗ctgf抗体抑制。将rCTGF与胶原凝胶一起注射到小鼠背部，在体内诱导了较强的血管生成。5) 3种细胞系（乳腺癌细胞系MDA231、纤维肉瘤细胞系HT1080和鳞状癌细胞系A431）中，MDA231产生CTGF的能力最高，A431最低，与它们在裸鼠体内形成血管生成肿瘤的能力平行。抗CTGF抗体抑制肿瘤诱导的血管生成。6)类风湿关节炎是一种血管生成疾病，在类风湿关节炎患者的滑液中存在CTGF。7) CTGF在实验性心肌梗死大鼠梗死区表达。8)在产生人型抗体的转基因小鼠体内培养人抗ctgf抗体。其中一种抑制CTGF分泌较多的MDA231肿瘤骨转移。这些结果表明，CTGF是一种新型的、有效的血管生成因子，在生理性和病理性血管生成的多个阶段发挥作用，提示抗CTGF抗体可作为血管生成抑制剂。

1) As inhibitors for connective tissue growth factor (CTGF), a polyclonal antibody was raised by immunizing a synthesis CTGF fragment into rabbit. In addition, monoclonal anti-CTGF antibodies were also prepared.2) Recombinant CTGF (rCTGF) promoted the adhesion, proliferation and migration of the vascular endothelial cells and these effects were inhibited by anti-CTGF antibody.3) rCTGF markedly induced the tube formation of vascular endothelial cells, and this effect was stronger than that of basic fibroblast growth of vascular endothelial growth factor.4) Application of rCTGF to the chicken chrioallantoic membrane (CAM) resulted in gross angiogenic response and the effect was inhibited by anti-CTGF antibody. rCTGF injected with collagen gels into the back of mice induced strong angiogenesis in vivo.5) Among three cell lines (breast cancer cell line MDA231, fibrosarcoma cell line HT1080 and squamous carcinoma cell line A431), the ability to produce CTGF was highest in MDA231 and lowest in A431 and was parallel to their ability to form angiogenic tumors in nude mice. Anti-CTGF antibody inhibited tumor-induced angiogenesis in CAM.6) CTGF was present in synovial fluid in patients with rheumatoid arthritis which is an angiogenic decease.7) CTGF was expressed in the infarct zone of experimentally induced myocardial infarction in rats.8) Human anti-CTGF antibodies were raised in transgenic mice producing human type antibody. One of them inhibited bone metastasis of MDA231 tumors which produce much CTGF.These findings indicate that CTGF is a novel, potent angiogenesis factor which functions in multi-stages in physiological and pathological angiogenesis and suggest that anti-CTGF antibody can be used as an inhibitor for angiogenesis.