佛手散治疗阿尔茨海默病分子机制的系统药理学研究  

An Insight into the Molecular Mechanism of Foshou Powder against Alzheimer’s Disease Based on Systems Pharmacology and Experimental Validation

作　　者：冯丽旋 郭鹏飞 周丽桂 谢清华 张永斌 郭学军 FENG Lixuan;GUO Pengfei;ZHOU Ligui;XIE Qinghua;ZHANG Yongbin;GUO Xuejun(Laboratory Animal Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学实验动物中心,广东广州510405

出　　处：《中药新药与临床药理》2021年第11期1622-1631,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省科学技术厅项目(2015A030302072);广州中医药大学2017年度“高水平大学建设”面上项目(A1-AFD018171Z11029)。

摘　　要：目的利用系统药理学方法和体外实验探究佛手散治疗阿尔茨海默病(Alzheimer disease,AD)的分子机制。方法利用TCMSP、TCM-Mesh、ETCM和TCMID数据库筛选佛手散的活性成分;利用TCMSP、ETCM和PharmMapper数据库对活性成分进行作用靶点预测。利用DisGeNet、GAD、CTD、TTD、HPO和Orphadata数据库筛选出AD疾病相关靶点,并与佛手散活性成分作用靶点映射取交集,交集基因即为佛手散治疗AD的潜在作用靶点。通过String数据库构建佛手散治疗AD潜在作用靶点的蛋白互作(PPI)网络;利用Cytoscape3.6.1软件构建"活性成分-潜在作用靶点"网络。通过WebSestalt及David分析平台对潜在作用靶点进行GO功能及KEGG通路富集分析,并进行体外实验验证。结果共筛选出佛手散52个活性成分,预测得到185个共同靶点,最终确定31个佛手散治疗AD的潜在作用靶点。GO功能富集分析结果表明,潜在作用靶点参与生物调控、刺激反应、细胞间通讯等生物学过程;细胞膜、蛋白复合物、囊泡参与等细胞组分;蛋白结合、离子结合、分子转导活性、水解酶活性等分子功能。KEGG富集分析得到13条信号通路。体外实验结果表明,佛手散对谷氨酸钠诱导损伤的PC12细胞有保护作用,可通过降低胞内Ca^(2+)浓度和NO含量达到降低细胞凋亡率的效果。结论佛手散在治疗AD方面具有多成分、多靶点、多途径的整体调控特点,可能与其通过豆甾醇、Z-藁本内酯、藁本内酯、β-谷甾醇等活性成分调控5-羟色胺能突触通路、神经配体-受体交互作用通路、Ca2+信号通路以及cGMP-PKG信号通路等多通路有关。Objective To explore the mechanism of Foshou powder in the treatment of Alzheimer’s disease(AD)by systematic pharmacology and in vitro experiments.Methods The active ingredients of Foshou powder were screened by TCMSP,TCM-Mesh,ETCM and TCMID database,and the targets of the active ingredients were predicted by TCMSP,PharmMapper and the MedChem Studio of ETCM database.The targets related to AD were screened by DisGeNet,GAD,CTD,TTTD,HPO and Orphadata database,and then the compound-target network was obtained by mapping AD related targets with active ingredients targets.Subsequently,candidate targets and components were screened by PPI network constructed using String database,and drug-target network was built using Cytoscape 3.6.1 software.Finally,GO analysis and KEGG enrichment analysis were carried out using WebSestalt and David tools,and verified by in vitro experiments.Results There were 52 active ingredients corresponding to 185 targets and finally narrowed down to 31 targets related to AD in Foshou powder.Results of enrichment analysis showed that the targets were involved in biological processes such as biological regulation,response to stimulus and cell communication;cellular component such as membrane,protein-containing complex,cell projection and vesicle participate;and molecular functions such as protein binding,ion binding,molecular transducer activity and hydrolase activity.KEGG enrichment analysis revealed 13 signaling pathways.In vitro experiments showed that Foshou powder could protect neurons by reducing the apoptosis,the concentration of Ca^(2+) and the content of NO in monosodium glutamate-induced PC12 cells.Conclusion Foshou powder has the characteristics of overall regulation by multi component-multi-target-multi-channel in the treatment of AD,acts on the serotonergic synapse pathway,neuroactive ligand-receptor interaction pathway,Ca2+signaling pathway,cGMPPKG signaling pathway,etc.,through stigmasterol,(Z)-ligustilide,ligustilide,sitosterol and other active ingredients.

关 键 词：佛手散 阿尔茨海默病 系统药理 钙离子 一氧化氮 神经细胞凋亡 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]