川芎嗪抑制与角膜新生血管增殖相关的趋化因子受体CXCR4的分子机制

角膜新生血管是一种严重角膜病变，其发生机制及防治一直是眼科领域的研究热点。趋化因子受体CXCR4在新生血管生成中起了重要作用，是目前治疗的靶基因。本课题组一直致力于该蛋白的研究，并发现：临床广泛应用的中药有效成分川芎嗪TMP能有效抑制多种细胞中CXCR4的表达，体内试验也证明川芎嗪能有效减少碱烧伤引起的角膜新生血管。但是川芎嗪如何调节CXCR4表达的分子机制目前尚不清楚。因此，本课题将在现有的研究基础上，以大鼠角膜碱烧伤模型为载体，通过高保真扩增CXCR4启动子序列、构建绿色荧光蛋白报告基因的调控质粒、体内转染、荧光检测等实验，确认CXCR4基因启动子的关键位点；继而采用DNA与蛋白结合、蛋白质双向电泳、质谱分析、CHIP等技术，筛选作用于CXCR4启动子的转录因子，从而阐明川芎嗪调节CXCR4表达的分子机制。该研究不仅能揭示角膜新生血管增生的本质，而且也能为川芎嗪的推广应用提供科学依据。

Corneal neovascularization (CNV) is one kind of serious corneal disease. The study of its mechanism, prevention and treatment is still hot topic in ophthalmology. Chemokine receptor, CXCR4, plays a key role in neovascularization. Our research projects have been related to this gene since 2007. Recently, we found TMP significantly down-regulated the expression of CXCR4 in many cells. And, TMP inhibits corneal neovascularization induced by alkali burn in vivo. However, the mechanism remains unclear. Therefore, our goals in this project are, ⑴ to determine the functional sites in CXCR4 promoter by transfecting the regulating plasmid with reporting gene, GFP in corneal neovascularization induced by alkali burn in vivo. ⑵ to discover the transcription factors of CXCR4 by DNA Pull-Down, Two-D page gel, MS and CHIP assay. This study will elucidate the mechanism of TMP treatment, which will extend its application internationally and give rise to greater impacts to traditional Chinese medicine.