基于网络药理学和动物实验探讨白头翁汤通过IL-23/IL-17轴对外阴阴道念珠菌病的治疗作用  

Exploring the therapeutic effects of Pulsatilla Decoction on Vulvovaginal candidiasis through the IL-23/IL-17 axis based on network pharmacology and animal experiments

作　　者：段强军 戴一[2] 黄茸茸[2] 赵颖 李静 陆松侠 DUAN Qiangjun;DAI Yi;HUANG Rongrong;ZHAO Ying;LI Jing;LU Songxia(School of Integrated Chinese&Western Medicine,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;School of Pharmacy,Anhui Xinhua University,Anhui Hefei 230088,China;Department of Pharmacy,Anhui Provincial Hospital of Integrated Traditional Chinese&Western Medicine,Anhui Hefei 230031,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽新华学院药学院,安徽合肥230088 [3]安徽省中西医结合医院药剂科,安徽合肥230031

出　　处：《中国医院药学杂志》2024年第13期1513-1519,共7页Chinese Journal of Hospital Pharmacy

基　　金：安徽省高校自然科学研究重点项目(编号:KJ2020A0424,KJ2020A0792);安徽中医药大学校级科学研究项目(编号:2020sfyzc02)。

摘　　要：目的:基于网络药理学预测和动物实验验证探讨白头翁汤对外阴阴道念珠菌病(Vulvovaginal candidiasis,VVC)的治疗作用及机制。方法:通过文献查阅及TCMSP、Gene Cards等数据库收集白头翁汤成分、靶点及VVC疾病靶点,Cityscape 3.6.0软件构建“药物活性成分-靶点-通路”网络;String数据库构建蛋白互作网络。通过DAVID平台进行基因功能(GO)分析、京都基因与基因组百科全书(KEGG)通路富集分析;使用AutoDock软件对核心成分及关键靶点进行分子对接。将SPF级KM小鼠采用随机数表法分为正常组,模型组,白头翁汤低、中、高剂量组,氟康唑组(n=12)。阴道腔内接种白色念珠菌建立VVC模型。连续给药14 d后,革兰氏染色法和平板计数法观察阴道灌洗液中真菌载量;HE染色观察阴道组织形态改变;ELISA法检测血清IL-17、IL-23、IL-1β的含量;Western blot法验证主要靶点TNF-α、HSP90、ESR、STAT3和IL-23/IL-17轴核心调控因子ROR-γT蛋白表达。结果:筛选得到白头翁皂苷B4、二氢尼洛替星、黄柏酮、β-谷甾醇、小檗碱为治疗VVC的核心成分;筛选出18个关键作用靶点和75条通路。动物实验结果显示,与模型组比较,白头翁汤给药后可降低阴道内真菌载量和血清炎症因子IL-17、IL-23和IL-1β含量,降低阴道组织内TNF-α、HSP90、ESR、STAT3和ROR-γT蛋白表达,差异具有统计学意义(P<0.05)。结论:白头翁汤通过调控STAT3、TNF-α等靶点以及IL-23/IL-17轴发挥治疗VVC作用。OBJECTIVE To explore the therapeutic effects and mechanism of Pulsatilla Decoction on Vulvovaginal candidiasis(VVC)based upon network pharmacology and animal experiments.METHODS The active ingredients and corresponding targets of Pulsatilla Decoction were collected through literature reviews and searching the database of TCMSP.Gene Cards was applied for obtaining the potential targets related to VVC and Cytoscape 3.6.0 for constructing the network of“drug active components-targets-pathways”.And the database of String was utilized for constructing a protein interaction network.Gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed by the platform of DAVID.Molecular docking was performed with AutoDock software.Specific pathogen free Kunming(SPF-KM)mice were randomized into six groups of normal,model,low/medium/high-dose Pulsatilla Decoction and fluconazole.VVC model was established through an inoculation of Candida albicans.All groups were continuously dosed for 14 days.Fungal quantities in vaginal lavage fluid were determined by Gram stain and plate counting.Hematoxylin-eosin(HE)stain was utilized for observing the morphology of vaginal tissue.Enzyme-linked immunosorbent assay(ELISA)was employed for detecting the serum contents of IL-17,IL-23 and IL-1β.The expressions of core regulatory factors,including TNF-α,STAT3,HSP90,ESR and key regulator ROR-γT protein in IL-23/IL-17 axis were measured by Western blot.RESULTS Through network pharmacology screening,the core components identified for treating VVC included anemoside B4,dihydroniloticin,obacunone,β-sitosterol and berberine.Additionally,18 key target proteins and 75 pathways were identified as crucial for therapeutic effects for VVC.As compared with model group,a treatment of Pulsatilla Decoction could significantly lower both fungal burden in vagina and serum levels of IL-17,IL-23 and IL-1β.Moreover,the protein expression levels of TNF-α,STAT3,HSP90,ESR and ROR-γT dropped in vaginal tissue(P<0.05).CONCLUSI

关 键 词：外阴阴道念珠菌病 白色念珠菌 白头翁汤 网络药理学 

分 类 号：R285[医药卫生—中药学] R96[医药卫生—中医学]