基于网络药理学及实验验证探讨萆苓止带方治疗外阴阴道假丝酵母菌病的作用机制  

Mechanism of Biling Zhidai Decoction on vulvovaginal candidiasis based on network pharmacology and experimental verification

作　　者：陈志霞[1,2,3] 陈思琪 冯淑仪 CHEN Zhixia;CHEN Siqi;FENG Shuyi(The Second Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510120,China;Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510120,China;Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome,Guangzhou 510120,China)

机构地区：[1]广州中医药大学第二临床医学院,广东广州510120 [2]广东省中医院,广东广州510120 [3]广东省中医证候临床研究重点实验室,广东广州510120

出　　处：《中草药》2024年第20期6995-7005,共11页Chinese Traditional and Herbal Drugs

基　　金：广东省基础与应用基础研究基金项目(2023A1515220004);广东省科技计划项目(2023B1212060063);广州市科技计划项目(2024A03J0119);国家中医药管理局——黄健玲全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号);广东省中医药局科研项目(20232058)

摘　　要：目的 采用网络药理学方法探究萆苓止带方治疗外阴阴道假丝酵母菌病(vulvovaginalcandidiasis,VVC)的作用机制,并进行体内实验验证。方法 运用TCMSID数据库获取萆苓止带方中活性成分,并预测药物作用靶点;运用GEO、MelaCards和CTD数据库获取VVC疾病基因,将萆苓止带方作用靶点与VVC疾病相关基因取交集。运用STRING在线工具构建蛋白质-蛋白质相互作用(protein-proteininteraction,PPI)网络,使用Cytoscape软件进行可视化处理;使用WebGestalt工具进行基因本体(gene ontology,GO)功能及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析;运用MCODE插件筛选关键基因,运用CB-DOCK2对关键靶点和成分进行分子对接。采用雌激素注射联合阴道接种白色念珠菌法制备VVC小鼠模型,给予萆苓止带方干预后,对阴道灌洗液进行涂片染色,观察阴道真菌载量;采用苏木素-伊红(hemato xylin-eo sin,HE)染色观察阴道组织病理观变化;采用免疫组化法检测阴道组织雄激素受体(androgen receptor,AR)表达。结果 萆苓止带方与VVC疾病的共有靶点共43个,涉及代谢、炎症调节、激素等相关生物过程,富集于氮代谢、雌激素信号传导通路、卵巢类固醇生成、缺氧诱导因子-1 (hypoxia-induciblefactor-1,HIF-1)信号通路、内分泌抵抗等信号转导通路。共有靶点网络的核心基因为AR、基质金属蛋白酶1 (matrixmetallopeptidase 1,MMP1)、聚二磷酸腺苷-核糖聚合酶1 [poly (adenosine diphosphate-ribose)polymerase 1,PARP1]和一氧化氮合酶3 (nitric oxide synthase 3,NOS3),核心子网络共3个,均与多种卵巢类固醇激素相关蛋白及其合成代谢、细胞炎症调控等密切相关。动物实验结果显示,萆苓止带方能够有效降低VCC小鼠阴道灌洗液中真菌载量,改善阴道黏膜病理,并下调阴道组织AR表达(P<0.001)。结论萆苓止带方治疗VVC的机制是多靶点、多通路的复杂过程,其机Objective To investigate the mechanism of Biling Zhidai Decoction(萆苓止带方)on treating vulvovaginal candidiasis(VVC)using network pharmacology method,and perform in vivo experiments for verification.Methods The effective components of Biling Zhidai Decoction were obtained from TCMSID database and targets were predicted;VVC disease genes were acquired from GEO,MelaCards and CTD databases,and the targets of Biling Zhidai Decoction with VVC disease-related genes were intersected.Protein-protein interaction(PPI)network was constructed using STRING online tool and visualized using Cytoscape software;Gene ontology(GO)function and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed using WebGestalt tool;MCODE plugin was used to screen core targets,CB-DOCK2 was applied for molecular docking between core targets and components.A VVC mouse model was prepared by combining estrogen injection with vaginal inoculation of Candida albicans.After intervention with Biling Zhidai Decoction,the vaginal lavage fluid was stained and the vaginal fungal load was observed;Hematoxylin eosin(HE)staining was used to observe pathological changes in vaginal tissue;Immunohistochemistry was used to detect the expression of androgen receptor(AR)in vaginal tissue.Results There were 43 common targets shared with Biling Zhidai Decoction and VVC disease,involving metabolism,inflammation regulation,hormones and other biological processes,enriching in nitrogen metabolism,estrogen signaling pathway,ovarian steroidogenesis,hypoxia-inducible factor-1(HIF-1)signaling pathway,endocrine resistance and other signaling pathways.The core genes in the common target network were androgen receptor(AR),matrix metallopeptidase 1(MMP1),poly(adenosine diphosphate-ribose)polymerase 1(PARP1)and nitric oxide synthase 3(NOS3),with three core sub-networks,all closely related to various ovarian steroid hormones and synthesis metabolism,cell inflammation regulation.Animal experiment results showed that Biling Zhidai Decoction could

关 键 词：萆苓止带方 外阴阴道假丝酵母菌病 网络药理学 雄激素受体 落新妇苷 京尼平苷酸 毛蕊花糖苷 23-乙酰泽泻醇B 23-乙酰泽泻醇C 芍药苷 白术内酯Ⅰ 白术内酯Ⅱ 白术内酯Ⅲ 绿原酸 槲皮苷 

分 类 号：R285.5[医药卫生—中药学]