文献类型：期刊文献

中文题名：基于网络药理学-分子对接及实验研究探讨当归效应组分治疗原发性痛经的作用机制

英文题名：Mechanism of Effective Components from Danggui(当归)in Treatment of Primary Dysmenorrhea:Based on Network Pharmacology,Molecular Docking and Experimental Verification

作者：王瑞琼[1,2];孙敏[1];吴国泰[1,2];王志旺[1,2];刘峰林[1];谢田朋[1];任远[1,2]

第一作者：王瑞琼

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中药药理与毒理学重点实验室,兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：39

期号：1

起止页码：49

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(编号:81660653);甘肃省青年博士基金项目(编号:2021QB-074)。

语种：中文

中文关键词：当归效应组分;原发性痛经;网络药理学;分子对接;作用机制

外文关键词：Effective components from Danggui(当归);Primary dysmenorrhea(PD);Network pharmacology;Molecular docking;Mechanism of action

摘要：目的:采用网络药理学-分子对接的方法,探讨当归效应组分治疗原发性痛经(PD)的物质基础及作用机制,并进行动物实验验证。方法:利用气相色谱-质谱联用仪(GC-MS)检测当归效应组分的化学成分;通过TCMSP、UniProt Database、Pubchem、Swiss Target Pridiction数据库检索当归效应组分化学成分的作用靶点;采用TTD、Drugbank、pharmGKB、Uniprot、DisGeNET数据库检索PD相关的靶点,并建立数据集;筛选成分靶点和疾病靶点映射的共同靶点;利用David和KOBAS数据库对共同靶点进行通路的富集与分析;采用Cytoscape软件构建“活性成分-靶点-通路”的可视化网络关系图,分析当归效应组分治疗PD的物质基础及作用机制;以Discovery Studio软件对网络模型分析结果进行分子对接验证;根据预测结果进一步进行实验验证:将72只雌性KM小鼠随机分为正常对照组、模型对照组、布洛芬0.07 g/kg组、丁苯酞0.05、0.10、0.20 g/kg组,每组12只。采用戊酸雌二醇联合缩宫素建立小鼠原发性痛经模型。观察丁苯酞对模型小鼠扭体行为的影响,检测血清中E_(2)、P、NO、PGF_(2α)含量及子宫组织中ER、PR蛋白表达。结果:分析并鉴定了当归效应组分中44种化学成分;检索得到对应靶点809个,映射得到与PD相关靶点53个,发现其中PTGS2、PTGS1、PPARG、UGT2B4、CA2、ALOX5、UGT2B7等13个靶点可能为当归效应组分治疗PD的重要潜在靶点;通路注释分析得到47条作用通路,其中花生四烯酸代谢、卵巢类固醇生成、类固醇激素生物合成、催产素信号通路、NF-κB信号通路等15条与PD相关;网络图结果显示,3-丁烯基苯酞、藁本内酯、β-金合欢烯、β-花柏烯、β-红没药烯、β-柏木烯等10个核心成分可能是当归治疗PD的重要药效物质;分子对接结果显示,3-丁烯基苯酞、藁本内酯与核心靶点度值前3位的靶蛋白(PTGS2、PTGS1、PPARG)有较好的结合性。动物实验结果显示,丁苯酞能延长原发性痛经模型小鼠扭体发生的潜伏期,降低扭体反应的发生次数,升高血清中NO、P含量而降低E2、PGF_(2α)的含量,下调子宫组织中ER蛋白表达而上调PR蛋白表达,ER/PR值下降(P<0.05或P<0.01)。结论:当归效应组分通过多成分、多靶点、多通路治疗PD,作用机制可能与激素调节、中枢镇痛、解痉、抗炎及能量代谢有关。
Objective:To explore the material basis and mechanism of effective components from Danggui(当)in treatment of primary dysmenorrhea(PD)by network pharmacology,molecular docking and experimental verification.Methods:The chemical constituents of effective components from Danggui against PD were determined by gas chromatography-mass spectrometry(CC-MS),and the targets of those chemical constituents were retrieved from TCMSP,UniProt,Pubchem and SwissTargetPrediction databases.The PD-related targets were obtained from TTD,Drugbank,pharmGKB,Uniprot and DisGeNET databases to establish a data set.The common targets of component and disease mapping were also screened out,and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of the common targets was performed by using David and KOBAS databases.The Cytoscape software was used to construct the“active ingredient-target-pathway"network to analyze the material basis and mechanism of effective components from Danggui in treatment of PD.Key targets and main active ingredients were selected for molecular docking by Discovery Studio.Based on the predicted results,experimental verification was carried out.Specifically,estradiol valerate combined with oxytocin was used to induce PD in mice.Then the 72 unfertilized female KM mice were random-Ly divided into a normal group,a model group,an ibuprofen group(o.07 g/kg),and high-,middle-and low-dose butylphthalide groups,12 mice in each group.We further observed the effects of butylphthalide on the writhing behavior of mice,serum levels of estradiol(E2),progesterone(P),nitric oxide(NO)and prostaglandin F2a(PGF2),and protein levels of estrogen receptor(ER)and progesterone receptor(PR)in uterus tissues.Results:A total of 44 active ingredients of effective components from Danggui were identified.Meanwhile,809 potential targets were predicted,and 53 PD-related targets were mapped.Further analyses found 13 key targets,including PTGS2,PTCS1,PPARG,UGT2B4,CA2,ALOX5 and UGT2B7 may be the potential targets of effective components from Danggui against PD.Among the 47 pathways obtained from pathway annotations,15 were associated with PD,such as arachidonic acid metabolism,ovarian steroidogenesis,steroid hormone biosynthesis,oxytocin signaling pathway and NF-kappa B signaling pathway.The network diagram results showed that 10 core components including 3-butylidenephthalide,ligustilide,β-farnesene,β-chamigrene,β-bisabolene andβ-cedrene may be the important effective substances of Danggui in treatment of PD.The results of molecular docking demonstrated that 3-butylidenephthalide and ligustilide had good binding ability to key proteins PTCS2,PTCS1 and PPARG.The results of animal experiments showed that butylphthalide can decrease the writhing times and prolong the latent period of PD mice,and it can also reduce E2 and PGF2e levels and increase P and NO levels in serum.Moreover,butylphthalide down-regulated ER levels and up-regulated PR levels in uterine tissues,and the ratio of ER/PR protein showed a downward trend.Conclusion:Effective components from Danggui can treat PD through multi-component,multi-target and multi-pathway mechanisms,which may be related to hormone regulation,central analgesia,spasmolysis,inflammation and energy metabolism.