Golubkov Vladislav, Hawes Debra, Markland Francis S
Department of Biochemistry, University of Southern California, Keck School of Medicine, Los Angeles, California 90033, USA.
Angiogenesis. 2003;6(3):213-24. doi: 10.1023/B:AGEN.0000021396.47009.b0.
Previous work in our laboratory has shown that contortrostatin (CN), a disintegrin from southern copperhead snake venom, possess anti-angiogenic activity. In the present study we further examined the anti-angiogenic activity of CN, focusing on the mechanisms of CN inhibition of angiogenesis. CN inhibited migration and invasion, and significantly altered Matrigel-induced tube formation of human umbilical vein endothelial cells (HUVEC), but did not affect cell viability, or MMP-2 and MMP-9 activity. Immunocytochemistry of HUVEC revealed that CN disrupted actin cytoskeleton and altered VE-cadherin distribution at cell-cell contacts. CN downregulated focal adhesion kinase (FAK) and paxillin tyrosine phosphorylation in adherent HUVEC. There was also significant inhibition of angiogenesis in vivo by CN as assessed by implanting Matrigel plugs in C57 mice and measuring ingrowth of blood vessels using either factor VIII staining or hemoglobin determination. In conclusion, the present findings confirm our earlier studies and demonstrate conclusively that CN possess strong anti-angiogenic activity in vitro and in vivo.
我们实验室之前的研究表明，来自南铜头蛇毒液的解整合素扭曲素（CN）具有抗血管生成活性。在本研究中，我们进一步研究了CN的抗血管生成活性，重点关注CN抑制血管生成的机制。CN抑制人脐静脉内皮细胞（HUVEC）的迁移和侵袭，并显著改变基质胶诱导的管腔形成，但不影响细胞活力，也不影响MMP-2和MMP-9活性。对HUVEC进行免疫细胞化学分析显示，CN破坏了肌动蛋白细胞骨架，并改变了细胞间连接处VE-钙黏蛋白的分布。CN下调了贴壁HUVEC中粘着斑激酶（FAK）和桩蛋白的酪氨酸磷酸化。通过在C57小鼠中植入基质胶栓子，并使用因子VIII染色或血红蛋白测定法测量血管向内生长，评估结果表明CN在体内也显著抑制了血管生成。总之，目前的研究结果证实了我们早期的研究，并确凿地证明CN在体外和体内均具有强大的抗血管生成活性。
