微脉冲阈值下光凝对离体和在体视网膜色素上皮细胞的光生物调制效应研究

病理状态下视网膜色素上皮细胞(RPE) 结构和功能的改变引发的黄斑病变是众多眼底病的病理学基础，可严重损害视功能，而常规激光光凝、药物、手术等众多治疗方法可促使眼底瘢痕形成、视力下降、药物副作用以及损害RPE和视网膜屏障功能，因此主动诱导RPE细胞增殖活化并分泌相应细胞因子对于视觉功能修复具有重要意义。鉴于微脉冲低能量激光对生物组织的光调制效应，本项目拟从离体培养RPE、正常大鼠在体RPE层和大鼠诱导CNV模型三个层次，利用免疫组化、RT-PCR、westernblot、高效液相色谱分析、视网膜电图等技术方法，探索微脉冲阈值下光凝如何刺激RPE产生活性氧和转录因子，触发下游生物学效应，抑制VEGF的表达，释放PEDF等血管生长抑制因子，同时分泌神经营养和生长因子，从而改善RPE结构和生物学功能，最终从分子水平揭示微脉冲低能量激光对RPE的生物调制效应，为黄斑部病变的治疗提供新思路、新方法。

Retinal pigment epithelia cells (RPE) constitue the most important part of retinal barrier. Under pathological state the structure and function of RPE changes a lot and initiates macular edema or macular choroidal neovascularization, which is the pathological basis of various ocular fundus diseases, then leads to severe visual impairment. Conventional laser photocoagulation, drugs and operations may increase the crisis of macular scar formation, impaired vision, drug toxic, side effect and retinal barrier impaiment. So, inducing RPE to reduce VEGF, promote PEDF, inhibti neovascularization, secrete neurotrophic factors and growth factors, facilitate the repair and hyperplasia of RPE is essential. In view of the photobiomodulation of micropulse low-power laser to RPE, this study plans to perform immunocytochemistry, RT-PCR, westernblot, HPLC, ERG and other molecular biological technologies to explore the effect of micropulse subthreshold photocoagulation on cultured RPE cells, normal rat RPE and laser induced rat CNV model. So to reveal the molecular mechanisms of biomodulation effect of micropulse low-power laser on RPE in vitro and in vivo, and to build new clues and strategies for macular lesions in future.