Melkonian G, Le C, Zheng W, Talbot P, Martins-Green M
Department of Neuroscience, University of California, Riverside, California, 92521, USA.
Toxicol Appl Pharmacol. 2000 Feb 15;163(1):26-37. doi: 10.1006/taap.1999.8789.
The adverse effects of cigarette smoke on mature blood vessels are well established, whereas little is known about the influence of smoke on blood vessel development. To determine if cigarette smoke alters angiogenesis, chick chorioallantoic membranes (CAMs) were exposed for 4 days to culture medium (control) or to mainstream (MS) or sidestream (SS) smoke solution, and then blood vessel patterns were compared in blind tests. In contrast to the normal tree-like branching of control blood vessels, smoke-treated CAMs often had vessels that ran parallel to each other without much branching. A significant increase in the number of fibroblasts was observed in histological sections of treated CAMs, and this increase correlated with alterations in extracellular matrix components. Many more matrix fibrils were observed in treated CAMs than in controls using scanning electron microscopy. Immunohistochemistry showed that type III collagen was distributed in a tight band adjacent to the endoderm in controls but was distributed throughout the mesoderm in both treatment groups. Western blots confirmed that both type I and type III collagen were more abundant in treated CAMs than in controls. Fibronectin, which was localized immunohistochemically in the basal laminae and mesodermal matrix of controls, increased in abundance in CAMs treated with SS smoke solutions. Hyaluronic acid, which was present in a dense band subjacent to the capillary plexus of control CAMs, was greatly reduced in MS-treated CAMs and was absent in SS-treated CAMs. These observations demonstrate that both MS and SS cigarette smoke solutions caused abnormal pattern formation of CAM blood vessels and altered the composition of the extracellular matrix in the CAM mesoderm.
香烟烟雾对成熟血管的不良影响已得到充分证实，而关于烟雾对血管发育的影响却知之甚少。为了确定香烟烟雾是否会改变血管生成，将鸡胚绒毛尿囊膜（CAMs）暴露于培养基（对照）、主流（MS）或侧流（SS）烟雾溶液中4天，然后在盲法试验中比较血管模式。与对照血管正常的树状分支不同，经烟雾处理的CAMs中的血管通常相互平行，分支较少。在经处理的CAMs组织切片中观察到成纤维细胞数量显著增加，且这种增加与细胞外基质成分的改变相关。使用扫描电子显微镜观察发现，经处理的CAMs中的基质纤维比对照中的多得多。免疫组织化学显示，III型胶原蛋白在对照中分布在内胚层相邻的紧密带中，但在两个处理组中均分布于整个中胚层。蛋白质免疫印迹证实，I型和III型胶原蛋白在经处理的CAMs中均比对照中更丰富。纤连蛋白在对照的基底膜和中胚层基质中免疫组织化学定位，在用SS烟雾溶液处理的CAMs中其丰度增加。透明质酸存在于对照CAMs毛细血管丛下方的致密带中，在经MS处理的CAMs中大大减少，在经SS处理的CAMs中则不存在。这些观察结果表明，MS和SS香烟烟雾溶液均导致CAM血管形成异常模式，并改变了CAM中胚层细胞外基质的组成。
