Maragoudakis M E, Haralabopoulos G C, Tsopanoglou N E, Pipili-Synetos E
University of Patras Medical School, Department of Pharmacology, Greece.
Microvasc Res. 1995 Sep;50(2):215-22. doi: 10.1006/mvre.1995.1054.
A method providing a biochemical index for the evaluation of promoters or inhibitors of angiogenesis in the chick chorioallantoic membrane (CAM) is here described and validated. This method is based on the determination of collagenous protein synthesis which takes place during new vessel formation. Validation was done by comparing collagenous protein synthesis to morphological methods of determining vascular density either by counting the number of vessels intersecting three concentric rings or by computer-assisted image analysis. Five compounds which promote or inhibit angiogenesis in the CAM were used for this purpose. The protein kinase C activator 4-beta-phorbol 12-myristate 13-acetate and alpha-thrombin increased collagenous protein synthesis and these results correlated with those obtained by using the two morphological methods mentioned above. Similarly, the inhibitors of angiogenesis, Ro318220, tricyclodecan-9-yl xanthate (D609), and 8,9-dihydroxy-7-methyl-benzo[b]quinolizinium bromide (GPA1734), reduced collagenous protein synthesis and vascular density (determined by image analysis or by counting the number of vessels intersecting three concentric rings) to a comparable degree. These results indicate that collagenous protein synthesis can be used as a reliable, reproducible, and unbiased index of angiogenesis in the chick chorioallantoic membrane.
本文描述并验证了一种为评估鸡胚绒毛尿囊膜（CAM）血管生成的促进剂或抑制剂提供生化指标的方法。该方法基于对新血管形成过程中发生的胶原蛋白合成的测定。通过将胶原蛋白合成与通过计数与三个同心环相交的血管数量或通过计算机辅助图像分析来确定血管密度的形态学方法进行比较来完成验证。为此使用了五种促进或抑制CAM血管生成的化合物。蛋白激酶C激活剂4-β-佛波醇12-肉豆蔻酸酯13-乙酸酯和α-凝血酶增加了胶原蛋白合成，这些结果与使用上述两种形态学方法获得的结果相关。同样，血管生成抑制剂Ro318220、三环癸烷-9-基黄原酸盐（D609）和8,9-二羟基-7-甲基苯并[b]喹啉溴化物（GPA1734）将胶原蛋白合成和血管密度（通过图像分析或通过计数与三个同心环相交的血管数量来确定）降低到相当程度。这些结果表明，胶原蛋白合成可作为鸡胚绒毛尿囊膜血管生成的可靠、可重复且无偏差的指标。
