运用网络药理学与分子对接法探讨阳和汤治疗股骨头坏死的核心作用机制

Applying network pharmacology and molecular docking in exploring the core mechanism of Yanghe Decoction in the treatment of femoral head necrosis

目的运用网络药理学与分子对接法,探究阳和汤(Yanghe Tang,YHT)治疗股骨头坏死(osteonecrosis of the femoral head,ONFH)的分子作用机制。方法通过中药系统药理学数据库(TCMSP数据库)、Uniprot数据库、GeneCards数据库和DAVID数据库,探究阳和汤方中有效化合物治疗ONFH的核心化合物、核心靶点和信号通路,并将YHT核心化合物与ONFH的重要靶点HIF-1a和血管内皮生长因子(vascular endothelial growth factor A,VEGFA)进行分子对接。结果通过TCMSP数据库搜索中药的化合物成分与对应靶点,依据口服吸收利用度(OB)≥30%,药物相似性(DL)≥0.18的筛选标准,去除重复值,最终检索到阳和汤方中主要活性化合物119个。靶点通过Uniprot基因校正后,得到226个靶点基因。通过GeneCards数据库共获得284个疾病靶点基因,将疾病靶点与化合物靶点交叉映射,进而获得阳和汤治疗ONFH的潜在作用靶点40个,并在STRING数据库中获得40个潜在作用靶点的PPI蛋白互作网络,通过Cytoscape 3.7.1软件对PPI网络进行拓扑分析,得到5个核心作用靶点:IL-6、TNF、TP53、VEGFA、CASP3。此外,与ONFH直接相关的重要靶点包括VEGFA、CASP3、HIF-1α、IL-6、TNF等。通过DAVID数据库,对潜在靶点进行GO功能注释和KEGG功能富集分析,最终获得与骨质代谢相关作用通路有PI3K-Akt信号通路,与免疫与炎症反应相关的通路有TNF信号通路,与血液供应作用相关的通路为HIF-1信号通路、VEGF信号通路、血管生成的正调控等。分子对接显示槲皮素与HIF-1α的结合能最低,豆甾醇与VEGFA的结合能最低,这两种分子对接方式更易发生。结论预测YHT可通过HIF-1信号通路、TNF信号通路、PI3K-Akt信号通路等途径,调控股骨头坏死进程中的免疫炎症反应、骨代谢和血液循环,达到干预和治疗ONFH的作用。

Objective To explore the molecular mechanism of Yanghe Decoction(Yanghe Tang,YHT)in thetreatment of femoral head necrosis(ONFH)by applying network pharmacology and molecular docking.MethodsThrough TCMSP database,Uniprot database,GeneCards database and DAVID database,the core compounds,core targets and signal pathways of effective compounds in Yanghe decoction in treaingt ONFH were probed,and YHT core compounds with ONFH's important target HIF-1a Molecular docking with VEGFA were compared.Results The TCMSP database was used to examine the compound components and corresponding targets of traditional Chinese medicine.According to the screening criteria of OB≥30%and DL≥0.18,duplicate values were removed,and finally 119 main active compounds in Yanghe Decoction were retrieved.After the target was corrected by Uniprot gene,226 target genes were obtained.A total of 284 disease target genes were obtained from the GeneCards database,and the disease targets and compound targets were cross-mapped to obtain 40 potential targets of Yanghe Decoction in the treatment of ONFH,and 40 potential targets in the STRING database.Through the Cytoscape 3.7.1 software,the PPI network was topologically analyzed,and five core targets(IL-6,TNF,TP53,VEGFA,and CASP3)were obtained in addition to important targets directly related to ONFH(including VEGFA,CASP3,HIF-1α,IL-6,TNF,etc).Through the DAVID database,the Go function annotation and KEGG function enrichment analysis of potential targets were performed,and finally the PI3K-Akt signaling pathway related to bone metabolism was obtained,and the TNF signaling pathway related to immune and inflammatory response was related to blood.The pathways related to supply were HIF-1 signaling pathway,VEGF signaling pathway,positive regulation of angiogenesis,etc.Molecular docking showed that the binding energy of quercetin and HIF-1αwas the lowest,and that of Stigmasterol and VEGFA was the lowest.These two molecular docking methods were more likely to occur.Conclusion It is predicted that YHT can regulate and control the immune inflammatory response,bone metabolism and blood circulation in the process of bone necrosis through HIF-1 signaling pathway,TNF signaling pathway,PI3K-Akt signaling pathway,etc.,and achieve the effects of intervention and treatment of ONFH.