基于复杂疾病数据库的视神经保护药物的作用机制研究

The protection of optic nerve is one of the essential treatments for its injury. Retinal ganglion cells (RGCs), retinal Müller cells (RMCs) and retinal endothelial cells (RECs) are selected as the subject in this study on the basis of our previous research work. The Chinese herbs improving blood circulation were collected from Chinese Pharmacopoeia and their chemical components were searched by Kyoto Encyclopedia of Genes and Genomes (KEGG) Database. The neuroprotective agents and drugs improving microcirculation of western medicine were appointed as the control chemicals.The genes relating optic atrophy were collected in Online Mendelian Inheritance in Man (OMIM), one of the classic Complex Diseases Databases. The gene expression differences will be compared by gene chip technology. According to the differences, we can build a network to reveal the functional connections between genes. Based on entries of Gene Oncology (GO) Database, we will categorize these by cell components, molecular function and biological processes of differentially expressed genes. The weighted calculation and arrangement of these effects will be set as the evaluation criterions to screen potential optic nerve protective components and combinations. The biochemical tests will be applied to prove neuroprotective effects. We will check the components and combinations above by its effects on the expression of specific markers, the secretion of nerve growth factor and small ubiquitin-related system in glucose-oxygen deprived RGCs, RMCs and RECs. Neuroprotective drugs and its combinations which proved having significant effects, definite composition and relatively clear action mechanism will be selected to lay the roots for progresses in optic nerve protective treatments.

视神经保护药物是治疗视神经损伤的重要途径。在前期工作基础上，本课题组以视网膜神经节细胞（RGCs）、Müller细胞和血管内皮细胞为研究对象，收集《中国药典》中活血植物药，从京都基因与基因组百科全书（KEGG）获取化学成分，以改善微循环和营养神经药物成分为对照，收集复杂疾病数据库在线人类孟德尔遗传（OMIM）与视神经萎缩相关基因，采用基因芯片技术检测这些成分造成的相关基因表达差异，构建网络揭示这些基因之间的功能联系；按照基因本体论（GO）数据库的注释分类差异表达基因的细胞组分、分子功能和生物学过程；根据这些化合物作用强度加权计算，筛选可能的视神经保护成分；以其对氧糖剥夺的三种细胞的特异标志物表达、神经生长因子分泌、小泛素修饰物系统的影响作为生物效应验证视神经和视网膜保护作用，从中优选作用强、机制相对明晰的视神经保护药物, 为按机制互补组合、提升临床视神经疾病的个体化治疗效果打下基础。

关于视神经疾病的发病机制研究已很多，但却未真正揭开全部致病机制的机理，视神经保护药物是治疗视神经损伤的重要途径。在前期工作基础上，本课题组以视网膜神经节细胞、Müller细胞和色素上皮细胞为研究对象，通过从京都基因与基因组百科全书完成了活血植物药及其化学成分的确定与收集；建立和鉴定大鼠视网膜神经节细胞、视网膜muller细胞、视网膜色素上皮细胞，并建立稳定的氧糖剥夺细胞体系；通过对视神经萎缩患者静脉血非编码RNA的表达测序，深入的生物信息学分析获得全转录组大数据；通过对差异基因进行分析，建立机制网络揭示这些基因之间的功能联系，生物化学方法进行验证，明确了视网膜色素上皮细胞和各种不同亚型视网膜神经节细胞对NMDA损伤的反应，miR-138调控大鼠视网膜神经节细胞、视网膜muller细胞、视网膜色素上皮细胞抗氧化应激作用的机制，靶向沉默SIRT1促进大鼠视网膜神经节细胞、视网膜muller细胞、视网膜色素上皮细胞凋亡；基于网络药理学的中药有效成分及潜在靶点的筛选，揭示视神经保护药物的作用机制。通过研究非编码RNA与视神经疾病的关系，网络药理学为手段寻找治疗视神经疾病靶点，从中优选作用强、机制相对明晰的视神经保护药物, 为按机制互补组合、提升临床视神经疾病的个体化治疗效果打下基础。在转录组学层面上为揭示视神经发病机制提供了一个新研究思路和方向，为视神经疾病防治和药物开发提供新作用靶点。