巩膜干细胞对实验性近视巩膜重塑的作用及调控机制研究

Myopia is a developmental oculopathy. Scleral remodeling is an important breakthrough in the etiology of experimental myopia. Type I collagen and cartilage proteins of sclera may be involved in the regulation mechanism of scleral remodeling. Stem cells are considered as a hot topic in the field of ophthalmology. Our prophase research found that adult stem cells with the potential abilities could be isolated from scleral tissue in guinea pig. Therefore, we hypothesize that scleral stem cells can regulate the process of scleral remodeling by regulating the expression of type I collagen and cartilage proteins. To test the hypothesize, (1) we will establish guinea pig experimental myopia model and simulate micro-environment of sclera remolding, then investigate the role of scleral stem cells in scleral remodeling related with experimental myopia by immunofluorescence, western-blot, RT-PCR and other molecular biological means. (2) We will regulate Wnt/β-catenin signal pathway by bidirectional intervention in vivo and vitro, and record the change of refractive error and axial length of guinea pig. We will confirm the regulation mechanism of scleral stem cells in scleral remodeling in experimental myopia with transmission electron microscopy and molecular biological means. We hope these researches will get some new findings that would lay the foundation for revealing the mechanism of experimental myopia, and bring new ideas of prevention and treatment of myopia.

近视是一种发育性眼病，巩膜重塑是实验性近视病因学研究的重要突破，研究认为I型胶原蛋白以及软骨相关蛋白参与调控巩膜重塑。干细胞作为眼科领域的研究热点，课题组前期研究发现，豚鼠巩膜组织可以分离出具有多向分化潜能的成体干细胞。为此，我们提出假说：豚鼠实验性近视模型巩膜干细胞通过影响I型胶原蛋白及软骨相关蛋白的差异性表达，继而调控巩膜重塑的发生发展。为验证以上假说，（1）构建豚鼠实验性近视模型以及模拟近视巩膜微观环境，采用免疫荧光、Western-blot、定量PCR等分子生物学手段，探讨巩膜干细胞对实验性近视巩膜重塑的作用；（2）通过体内外双向干预Wnt/β-catenin信号通路，观察豚鼠近视屈光度及眼轴变化，采用透射电镜技术观察和分子生物学手段明确巩膜干细胞对实验性近视巩膜重塑的调控机制。本项目将以巩膜干细胞为新视角，为揭示实验性近视发生机制奠定基础，为近视防治提供新的思路。