VITRONECTIN RECEPTOR ON SKIN ENDOTHELIAL CELLS

损伤、炎症和血管渗漏导致血管外 血清基质蛋白沉积。 微血管内皮细胞 利用整合素受体，特别是玻连蛋白受体（VnR）， 与沉积的基质结合，并将其用作形成新血管的支架。 的 VnR是调节血管内皮细胞功能的关键因素， 伤口愈合因为它可以结合多种蛋白质如纤维蛋白原， 玻连蛋白、凝血酶、血小板反应蛋白和血管性血友病因子（vWF） 其特征性地沉积在受损组织中。 初步 愈合皮肤伤口的研究表明，VnR的表达是 在创伤后浸润细胞上迅速增加， 其他整合素受体的表达。 然而，监管 微血管内皮细胞VnR的表达及其功能 在伤口愈合、炎症或损伤的背景下， 考察 因此，这项建议的具体目的是：1） 表征α（v）/β（3）复合物（玻连蛋白）的表达 人真皮微血管内皮细胞（HDMEC）上的VnR 使用免疫荧光显微术，流式细胞术，蛋白质免疫印迹， 和免疫沉淀。 2)为了检验可溶性因子的影响， 碱性成纤维细胞生长因子（FGF）、转化生长因子β（TGF-β） β）和蛋白激酶C激动剂对VnR表达的影响。 3)到 确定HDMEC与纯化的确定基质蛋白的相互作用， 检查这些是否被改变表达的试剂调节 的VNR。 4)为了确定亚基基因在分子水平上 表达和复杂的组装和表面表达是组成性的 调节并表征碱性FGF、PKC激动剂和 TGF β对调节机制的影响。 这些研究将产生新的 深入了解伤口愈合中内皮细胞的功能， 新生血管形成

Injury, inflammation and vascular leakage results in extravascular deposition of serum matrix proteins. Microvascular endothelial cells utilize integrin receptors, particularly the vitronectin receptor (VnR), to bind to deposited matrix and use it as a scaffold to form new vessels. The VnR is a crucial element in the regulation of endothelial cell function in wound healing since it can bind to multiple proteins such as fibrinogen, vitronectin, thrombin, thrombospondin, and von Willebrand's factor (vWF) which are characteristically deposited in injured tissues. Preliminary studies of healing skin wounds suggest that the expression of the VnR is rapidly increased on infiltrating cells after wounding, prior to increases in the expression of other integrin receptors. However, the regulation of expression and function of the VnR on microvascular endothelial cells within the context of wound healing, inflammation, or injury has not been examined. Therefore, the specific aims of this proposal are: 1) To characterize the expression of the alpha(v)/beta(3) complex (vitronectin receptor, VnR) on human dermal microvascular endothelial cells (HDMEC) using immunofluorescence microscopy, flow cytometry, western immunoblot, and immunoprecipitation. 2) To examine the effects of the soluble factors basic fibroblast growth factor (FGF), transforming growth factor beta (TGF- beta), and protein kinase C agonists on the expression of the VnR. 3) To define the interaction of HDMEC with purified defined matrix proteins and examine whether these are modulated by agents which change the expression of the VnR. 4) To determine the molecular levels at which subunit gene expression and complex assembly and surface expression are constitutively regulated and to characterize the effects of basic FGF, PKC agonists, and TGF beta upon regulatory mechanisms. These studies will produce novel insights into endothelial cell function in wound healing and neovascularization.