调经汤经铁死亡途径干预早发性卵巢功能不全的网络药理靶点分析及活性成分预测

Network pharmacological target analysis and active component prediction of Tiaojing decoction intervention in premature ovarian insufficiency by ferroptosis pathway

目的通过网络药理学及生物信息学方法,探索临床效方调经汤通过铁死亡途径治疗早发性卵巢功能不全(premature ovarian insufficiency,POI)的有效靶点和活性药物成分预测。方法通过中药系统药理学分析平台、用于传统中药分子机制挖掘生物信息学分析工具数据库获取调经汤的活性成分和靶基因,使用人类基因综合数据库、药物靶点数据库、Drugbank获取POI的疾病靶点,并通过Venn获取交集靶点。利用铁死亡数据库、Cytoscape及STRING软件构建调经汤通过铁死亡途径治疗POI的核心靶基因网络。使用DAVID数据库对调经汤治疗POI和铁死亡相关的预测靶点进行基因本体和生物学途径富集分析。通过基因表达总表(gene expression omnibus,GEO)数据库获取POI疾病生物学差异基因进一步提取关键靶点进行分析,并通过分子对接验证活性小分子药物成分结合力。结果调经汤13种中药中共筛选出154种活性成分,与POI相关靶点101个,铁死亡相关基因共23个。富集分析显示主要涉及通路为磷脂酰肌醇3-激酶-蛋白激酶B(phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B,PI3K/Akt)信号通路、低氧诱导因子-1信号通路等。通过GEO数据库筛得调经汤通过铁死亡途径干预POI的关键靶点为糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK-3β)、小窝蛋白-1(caveolin-1,CAV1)、哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)和蛋白激酶Cα(protein kinase C alpha gene,PRKCA),这些基因在POI疾病中表达量均降低。分子对接结果显示CAV1-槲皮素、GSK-3β-木犀草素具有较稳定的结合能力。结论网络药理学结果提示铁死亡途径可能是调经汤治疗POI的重要机制,GSK-3β、CAV1、mTOR和PRKCA等靶点及PI3K/Akt信号通路或在其中发挥重要作用。

Objective To explore the effective target and active ingredient prediction of premature ovarian insufficiency(POI)treated by Tiaojing decoction by ferroptosis pathway by network pharmacokogy and bioniformatics.Methods The active ingredients and target genes of Tiaojing decoction were obtained through traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine databases,disease targets for POI were obtained by using GeneCards,therapeutic target database and Drugbank,and intersection targets were obtained by Venn.Core target gene network of Tiaojing decoction in treatment of POI were constructed through the ferroptosis pathway by ferroptosis database,Cytoscape and STRING software.Then,the DAVID database was used to perform gene ontology and biological pathway enrichment analysis on the predicted targets related to ferroptosis of POI treated with Tiaojing decoction.Subsequently,differential genes in biology related to POI were obtained through the gene expression omnibus(GEO)database for further extraction of key targets,and the binding affinity of active small molecule drug components were verified through molecular docking.Results A total of 154 active ingredients,101 targets related to POI and 23 genes related to ferroptosis were selected from 13 kinds of traditional Chinese medicines in Tiaojing decoction.The enrichment analysis showed that the main involved pathways were phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B(PI3K/Akt)signaling pathway and hypoxia inducible factor-1 signaling pathway.Through GEO database screening,the key targets of Tiaojing decoction in intervening POI through ferroptosis pathway were glycogen synthase kinase-3β(GSK-3β),caveolin-1(CAV1),mammalian target of rapamycin(mTOR)and protein kinase C alpha gene(PRKCA),all expression of which reduced in POI.The results of molecular docking showed that CAV1-quercetin and GSK-3β-luteolin had stable binding ability.Conclusion The network pharmacology results suggest that the ferroptosis pathway may be an important mechanism of Tiaojing decoction in treatment of POI,GSK-3β,CAV1,mTOR,PRKCA and other targets,as well as PI3K/Akt signaling pathway,may play important roles in them.