基于网络药理学和实验验证探讨二仙汤及其温肾拆方治疗抑郁症的可行性  

Feasibility of Erxian Decoction and Wenshen Prescription in Treatment of Depression Based on Network Pharmacology and Experimental Verification

作　　者：佘楷杰 巩子汉 杨婧雯 梁文青 孟丹华 岳广欣[1] SHE Kaijie;GONG Zihan;YANG Jingwen;LIANG Wenqing;MENG Danhua;YUE Guangxin(Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Basic Medicine,Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]中国中医科学院中医基础理论研究所,北京100700 [2]河南中医药大学基础医学院,河南450000

出　　处：《中国实验方剂学杂志》2022年第16期211-220,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(82174251,81573846);中国中医科学院科技创新工程项目(CI2021A00607);中央级公益性科研院所基本科研业务费专项(YZ-202107,YZ-202153)。

摘　　要：目的:基于网络药理学技术预测二仙汤全方和温肾方治疗抑郁症的分子机制,通过母婴分离结合慢性束缚应激抑郁模型进行药效及机制对比,探讨温肾拆方治疗抑郁症的可行性。方法:通过中药系统药理学数据平台(TCMSP)和中药分子机制生物信息学数据库(BATMAN)收集二仙汤全方及温肾方的活性成分及作用靶点;利用人类基因数据库(Genecards)、在线人类孟德尔遗传数据库(OMIM)、药物银行(Drugbank)等数据库筛选抑郁症相关靶点,与药物靶点取交集获得药物-疾病共同靶点,随后导入Cytoscape 3.8.2软件绘制中药-活性成分-靶点-疾病网络图;利用STRING平台构建蛋白互相作用网络并筛选核心靶点及关联的核心成分;采用Metascape平台对交集靶点进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)功能富集分析。采用母婴分离结合慢性束缚应激制备抑郁小鼠模型,在离乳第21天(PD21)至束缚完成第111天(PD111)给予二仙汤全方和温肾方的药混饲料进行干预。根据糖水偏好实验、悬尾实验、旷场实验、高架O迷宫实验评估小鼠抑郁状态;免疫组织化学法(IHC)观察小胶质细胞离子钙接头蛋白-1(Iba-1)表达;蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)检测蛋白激酶B1(Akt1)、脑源性神经营养因子(BDNF)、突触后致密物95(PSD95)、突触素(Syn)等表达水平。结果:共筛选二仙汤全方和温肾方治疗抑郁症靶点126和118个,全方仅多8个靶点。两方核心靶点相同,主要包括Akt1、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、KEGG通路富集分析预测二仙汤全方和温肾方治疗抑郁症主要涉及磷脂酰肌醇3-激酶(PI3K)/Akt信号通路、丝裂原活化蛋白激酶(MAPK)信号通路及神经活性配体-受体相互作用通路。动物实验表明,与抑郁症模型组比较,二仙汤全方和温肾方均可明显�Objective:To predict the molecular mechanism of Erxian decoction and Wenshen prescription(modified Erxian decoction)in the treatment of depression based on network pharmacology and explore the feasibility of Wenshen prescription in the treatment of depression by comparing the efficacy and mechanism of the two decoctions based on a depression model induced by maternal separation combined with chronic restraint stress.Method:Active components and targets of Erxian decoction and Wenshen prescription were collected through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Bioinformatics Analysis Tool for Molecular mechanism of Traditional Chinese Medicine(BATMAN-TCM).Targets related to depression were screened out from databases such as GeneCards,Online Mendelian Inheritance in Man database(OMIM),and DrugBank.Common targets of drugs and disease were obtained and imported to Cytoscape 3.8.2 to plot the drug-active component-target-disease network.STRING platform was used to construct a protein-protein interaction(PPI)network and core targets and related core components were screened out.Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment analysis were performed on common targets through Metascape platform.The depression model was induced in mice by maternal separation combined with chronic restraint stress.From the 21st day of maternal separation(PD21)to the 111th day of restraint stress completion(PD111),mice were fed with the diet mixed with Erxian decoction or Wenshen prescription for intervention.The depressive state of mice was evaluated according to the sucrose preference test,tail suspension test,open field test,and elevated O-maze test.The expression of ionized calcium-binding adapter molecule 1(Iba1)in the microglia was observed by immunohistochemistry(IHC).Western blot and Real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR)were used to detect the expression levels of protein kinase

关 键 词：抑郁症 二仙汤 拆方研究 网络药理学 实验验证 

分 类 号：R2-0[医药卫生—中医学] R33R289