血管形成和内皮细胞增殖在葡萄酒色斑病程进展中的作用及其机制研究

Port-wine stain (PWS) is a congenital capillary malformation that occurs in 0 3% of newborns. PWS lesions are usually flat and pink at the beginning, and become nodular, hypertrophic, or darker with age. The majority of PWS (70%) are located on the face and neck area and therefore can cause physical and psychosocial problems. . Capillary malformation was not been classified as hyperplasia of cells in the past, but the expansion of malformed tissues. However, in previous studies about the expression of vascular epithelial growth factor in port-wine stain，we can obviously find up-regulated expression of it. And several literatures report that the microscopic pathology shows the proliferation of a large number of endothelial cells in nodular port-wine stain. VEGF is considered to be one of the most important angiogenesis regulation factor,If we can intensively study how angiogenesis and the proliferation of endothelial cells by VEGF influence the hyperplasia and reconstruction of the lesion of port-wine stain, the mechanism of the incrassation of port-wine stain may be further explained and it can also provide theoretical foundation for the prevention and treatment of port-wine stain, and the development of some biomarkers. Our project will first base on the culture of endothelial cells in the laboratory combined with pathology, intensively study the biological property of endothelial cells in port-wine stain and the cellular and molecular mechanism of angiogenesis with VEGF.

葡萄酒色斑（port-wine stain，PWS）为先天性毛细血管畸形。随年龄增长，患者可出现病灶颜色的加深，增厚及结节生成，并可伴有累及区域的软组织及骨骼肥大，继而造成外观及功能上的障碍，严重影响患者身心健康。既往认为毛细血管畸形在本质上多数不是细胞增生，而是畸形组织的扩张。然而，在过去对葡萄酒色斑进行血管新生调节因子表达的研究中，可见明显表达上调。另有多篇国内外报道发现结节型葡萄酒色斑其镜下病理可见结节内大量内皮细胞增殖现象。VEGF被认为是最重要的血管生成调节因子，若能深入研究其在葡萄酒色斑病灶增生及重建中对血管形成，内皮细胞增殖的影响及作用机制，则可进一步阐明葡萄酒色斑增厚机制，为葡萄酒色斑病程发展的防治及某些生物标志物的开发提供一定的理论基础。本课题将以前期实验培养葡萄酒色斑内皮细胞结合病理为基础，深究VEGF在葡萄酒色斑内皮细胞生物学特性及血管形成分子细胞学机制。

【背景与目的】 葡萄酒色斑（port-wine stain，PWS）为先天性毛细血管畸形。随年龄增长，患者可出现病灶颜色的加深，增厚及结节生成，并可伴有累及区域的软组织及骨骼肥大，继而造成外观及功能上的障碍，严重影响患者身心健康。葡萄酒色斑目前已被明确定位是由 GNAQ 中一种体细胞激活突变引起，从而引起下游信号转导通路调节异常，发生血管畸形。本旨在通过组织病理分型，血管生成调节因子及相关基因测序，进一步阐明PWS增厚的发病机制。.【材料与方法】 对临床增厚及结节PWS病灶进行组织病理学分型，采用免疫组化法测定不同类型增厚结节PWS其血管生成调节因子VEGF，MMP-9，ANG-2，FGF-2的表达水平，并进行GNAQ基因测序及相关性分析。.【结果】 通过组织病理学研究发现葡萄酒色斑增厚结节的两种分型：血管扩张型及内皮细胞增殖型；增厚型PWS病灶既可表现为大量毛细血管畸形扩张，管壁增厚，形成后微静脉样畸形血管；也可表现为血管管腔明显增多，血管内皮细胞显著增殖的毛细血管瘤表现。据此，免疫组化VEGF，MMP-9，ANG-2，FGF-2检验发现镜下细胞增殖旺盛的“细胞增殖型”结节PWS其VEGF，MMP-9，ANG-2，FGF-2表达上调明显，而“血管扩张型”增厚PWS及正常对照皮肤其VEGF，MMP-9，ANG-2，FGF-2表达均无明显上调；及增厚PWS与GNAQ基因突变的显著相关性。.【结论】 临床增厚结节PWS可能由两种不同途径的发病机制引起，导致不同病理类型的构成：以血管明显扩张为主的“血管扩张型”增厚PWS，以及以内皮细胞大量增殖为主的“细胞增殖型”结节PWS；其中，内皮血管生长因子的上调可能在“细胞增殖型”结节PWS的发生发展中起重要作用；GNAQ基因突变可能是导致先天肥厚PWS病灶的重要发病机制。