枸杞子促进虹膜色素上皮细胞定向分化治疗视网膜色素变性的分子网络调控机制研究  

Molecular network regulation mechanism of Lycii Fructus in promoting directional differentiation of iris pigment epithelial cells to treat retinitis pigmentosa

作　　者：宋厚盼[1,2] 曾梅艳 陈小娟 陈新怡 彭俊 周亚莎 杨毅敬 杨焘 蔡雄 彭清华[1,3] SONG Houpan;ZENG Meiyan;CHEN Xiaojuan;CHEN Xinyi;PENG Jun;ZHOU Yasha;YANG Yijing;YANG Tao;CAI Xiong;PENG Qinghua(Hunan Provincial Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, Changsha 410208, Hunan,China;College of TCM, Hunan University of Chinese Medicine, Changsha 410208, Hunan,China;Medical College, Hunan University of Chinese Medicine, Changsha 410208, Hunan,China)

机构地区：[1]湖南中医药大学中医诊断学湖南省重点实验室,湖南长沙410208 [2]湖南中医药大学中医学院,湖南长沙410208 [3]湖南中医药大学医学院,湖南长沙410208

出　　处：《中国临床药理学与治疗学》2019年第12期1402-1414,共13页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目(81804150,81703920);中国博士后科学基金资助项目(2019M662790);湖南省自然科学基金项目(2019JJ50442,2019JJ40226);中医药防治眼耳鼻咽喉疾病湖南省重点实验室开放基金项目(2018YZD05);湖南中医药大学中医学一流学科开放基金项目(2018ZYX26,2018ZYX20)

摘　　要：目的:基于生物信息学和网络药理学方法探究枸杞子(LF)促进虹膜色素上皮细胞定向分化治疗视网膜色素变性(RP)的分子网络调控机制。方法:在中药系统药理学数据库和分析平台(TCMSP)检索LF的主要活性成分及其可能的作用靶点;从GEO数据库下载GSE81058基因表达谱数据,采用R软件构建虹膜色素上皮细胞(IPE)和视网膜色素上皮细胞(RPE)差异表达基因火山图和聚类热图;采用STRING数据库构建LF成分-靶点和IPE-RPE差异表达基因的蛋白质-蛋白质交互网络(PPI),运用Cytoscape提取交集网络基因;通过DAVID数据库对交集基因进行基因本体和信号通路分析;采用cytoHubba分析LF作用于IPE的关键靶点。结果:共获得LF的化学成分188个,其中主要的入血活性成分36个,这些活性成分可能作用的靶点201个;IPE与RPE主要的差异表达基因有142个,其中上、下调基因各71个;发掘得到105个与LF促进IPE分化以治疗RP的相关基因靶点;LF作用于这些靶点涉及的生物学过程包括调节细胞分化、调节视网膜层形成等,相关的分子功能主要涉及转录共激活剂活性、离子通道活性等,主要富集于细胞质、高尔基体、细胞核等区域;LF作用于IPE涉及的分子机制包括调控PI3K-Akt信号通路、神经活性配体-受体相互作用通路等;LF促进IPE分化主要作用于SPP1、MMP9等十个关键靶点。结论:IPE与RPE具有明显差异的基因表达谱,LF的有效活性成分可作用于这些差异表达基因,促进IPE细胞分化为RPE,从而治疗RP。本研究可为细胞移植与中医药联合的中西医结合疗法应用于RP的临床治疗提供科学依据。AIM:To investigate the molecular network regulation mechanism of Lycii Fructus(LF)in promoting directional differentiation of iris pigment epithelial cells in the treatment of retinitis pigmentosa(RP)based on the methods of bioinformatics and network pharmacology.METHODS:The main active ingredients and potential targets of LF were retrieved from TCMSP database and analysis platform.The data of GSE81058 gene expression profile was downloaded from GEO database.Volcanic maps and clustering thermal maps of differentially expressed genes between IPE and RPE were plotted by ggplot2 and Pheatmap packages,respectively.Protein-protein interaction(PPI)networks of LF component-target and IPE-RPE differentially expressed genes were constructed by STRING database.Cytoscape software was used to extract genes of the intersection network.Gene ontology and signaling pathway analysis of intersection genes were carried out through DAVID online database.CytoHubba was used to analyze the key targets of LF.RESULTS:A total of 188 chemical constituents of LF were obtained,of which 36 were the main active ingredients in blood.There were 201 possible targets for these active ingredients.142 differentially expressed genes of IPE and RPE were obtained,of which 71 were up-regulated and 71 were down-regulated.105 gene targets related to LF promoting IPE differentiation and treating RP were identified.The biological processes involved in LF acting on these targets included regulating cell differentiation and retinal layer formation.The related molecular functions mainly involved transcriptional co-activator activity,ion channel activity,etc.They were mainly concentrated in cytoplasm,Golgi apparatus,and other areas.The molecular mechanisms involved the regulation of PI3K-Akt signaling pathway,neuroactive ligand-receptor interaction pathway,etc.The key targets of LF for IPE differentiation include SPP1,MMP9,SNAI2,etc.CONCLUSION:The gene expression profiles are of significant differences between IPE and RPE.The active ingredients of LF can act on

关 键 词：视网膜色素变性 生物信息学 网络药理学 虹膜色素上皮细胞 视网膜色素上皮细胞 枸杞子 

分 类 号：R965.2[医药卫生—药理学]