色素上皮衍生因子对角膜损伤后神经再生的保护作用研究

Corneal injury is the common causes of blindness in Ophthalmology, photorefractive keratectomy also causes damages of corneal nerve. Since nerve regeneration after injury is crucial to corneal prognosis, study of corneal nerve regeneration and its pretection after injury has important scientific and application prospect. This project studies the neuroprotection role of PEDF from two aspects as in vivo and in vitro: (1) by Western Blot and immunofluorescent staining techniques, using corneal stromal cell transforming culture model and corneal epithelial cell injuried-cornea tissue culture model, study the changes of local PEDF synthesis and secretion after corneal injury; using trigeminal ganglion neurons culture model, study the effect of PEDF on trigeminal ganglion neurons and its cell membrane receptors; (2) by in vivo confocal microscopy and immunofluorescence staining technique, using two rabbit models of corneal lamellar and nerve injury, from two aspects of corneal nerve function recovery and nerve morphologic recovery, study the protective effect of exogenous PEDF on corneal nerve regeneration after injury. This study is aimed to explore the change of endogenous PEDF in the course of corneal injury, the mechanism and effect of exogenous PEDF on corneal nerve regeneration after injury, and to provide scientific basis and new thinking for the clinical development of new drugs on corneal nerve regeneration and protection.

角膜损伤是眼科常见致盲原因，屈光性角膜成形手术也同时损伤角膜神经。损伤后神经再生攸关角膜预后，研究角膜损伤后神经再生及其保护具有重要的科学和应用前景。本课题从体内和体外两方面对PEDF神经保护作用进行研究：(1)应用Western Blot和免疫荧光染色技术，采用角膜基质细胞转化培养模型和角膜上皮细胞损伤的角膜组织培养模型，研究角膜损伤与局部PEDF生成与分泌的变化；采用三叉神经节神经元细胞培养模型，研究PEDF对三叉神经节神经元细胞的作用及其细胞膜受体情况；(2)应用活体共焦显微镜和免疫荧光染色技术，采用两种兔角膜浅层及神经损伤的动物模型，从角膜神经功能和神经形态学恢复两方面，研究外源性PEDF对角膜损伤后神经再生的保护作用。本课题旨在探讨角膜损伤过程中内源性PEDF的变化和外源性PEDF在角膜神经损伤再生中的作用机制和效果，为临床开发新的角膜神经再生保护药物提供科学依据和新思路。

角膜损伤是眼科常见致盲原因，屈光性角膜成形手术也同时损伤角膜神经。损伤后神经再生攸关角膜预后，研究角膜损伤后神经再生及其保护具有重要的科学和应用前景。本课题从体内和体外两方面对PEDF 神经保护作用进行研究：(1)应用RT-PCR、Western Blot 和免疫荧光染色技术，采用角膜基质细胞转化培养模型和角膜上皮细胞损伤的角膜组织培养模型，研究角膜损伤与局部PEDF 生成与分泌的变化；初步探讨PEDF促进神经修复的细胞信号通路； (2)应用活体共焦显微镜和免疫荧光染色技术，在兔角膜激光层间切开, 神经损伤的动物模型上，从角膜形态学恢复方面，研究外源性PEDF 对角膜损伤后神经再生的保护作用。结果显示， 角膜上皮损伤后局部PEDF的分泌减少， 同时加剧了炎症反应，COX2表达增高；PEDF mRNA 的表达在不同时间段和不同处理组间均有不同表达；初步得出PEDF促进损伤后的角膜神经修复是通过提高损伤后的角膜基质细胞15LOX的合成使得neuroprotectin D1(NPD1)的合成增加，从而促进神经修复；外源性的PEDF可促进兔角膜神经损伤的修复。