基于网络药理学和分子对接探讨高频调髓中药治疗股骨头坏死的作用机制

Mechanism of Chinese Herbs of Regulating Marrow in the Treatment of Osteonecrosis of the Femoral Head Based on Network Pharmacology and Molecular Docking

运用网络药理学和分子对接方法,探讨熟地、牛膝、茯苓、肉苁蓉和菟丝子等高频调髓中药治疗股骨头坏死(Osteonecrosis of the femoral head,ONFH)的作用机制。借助TCMSP平台,获取高频调髓中药的活性成分与潜在作用靶点,将上述药物作用靶点在Uniprot数据库中进行标准化处理。通过GeneCards和OMIM两大疾病数据库,获取得到ONFH疾病相关基因靶点,对所得到的已规范化的药物活性成分与疾病靶点进行整理合并,得到交集基因靶点。通过Cytoscape软件和String数据库,绘制药物-活性成分-靶点-疾病网络和构建蛋白互作网络,分析网络中关键活性成分和蛋白靶点。利用R软件对交集基因进行基因本体(Geneontology,GO)功能和京都基因与基因组百科全书(Kyoto Encyclopediaof Genesand Genomes,KEGG)通路富集分析,得到交集基因富集的功能与信号通路。为了验证药物活性成分与作用靶点结合的稳定性,使用AutoDockTools和AutoDock Vina软件进行分子对接验证,并在PyMOL软件中进行可视化分析,共得到5味调髓中药54个活性成分,1074个药物靶点,195个作用基因靶点,ONFH相关疾病靶点1219个。调髓中药有效作用靶点与疾病交集靶点39个。GO功能分析得到1619个条目,包括1517个生物过程,10个细胞组成及92个分子功能,其主要与血管生成、骨、脂质代谢和炎症反应等过程相关。KEGG通路富集分析得到119条通路,主要有AGE-RAGE信号通路、PI3K-Akt信号通路和IL-17信号通路等。分子对接结果显示,槲皮素、汉黄芩素和山柰酚等关键活性成分与IL6、P53和VEGF等核心蛋白具有良好的亲和力。本研究表明,高频调髓中药可通过调节免疫炎症反应、血管生成和骨代谢等过程,而达到治疗ONFH的作用,其具有多成分、多靶点和多途径的综合治疗作用特性。

This paper discussed the mechanism of high-frequency marrow-regulating Chinese medicines,such as Shudi,Niuxi,Fuling,Roucongrong,and Tusizi,in treating osteonecrosis of the femoral head(ONFH) by using network pharmacology and molecular docking methods.With the TCMSP,the active ingredients and potential targets of high-frequency Chinese herbs of controlling marrow were discovered,and the UniProt database was used to standardize the drug targets.The GeneCards database and the OMIM database were used to find the genes that were targeted for the ONFH.The standardized drug targets and disease targets were intersected to get the intersecting targets.Using Cytoscape software and the STRING database,the drug-component-target-disease network and protein-protein interaction network were built,and the key active ingredients and targets were examined.In order to investigate the function and signal pathway of the enrichment of overlapping genes,the gene ontology(GO) and the Kyoto Encyclopediaof Genesand Genomes(KEGG) were analyzed by R software.Finally,in order to establish the stability of the combination of drug active components and action targets,molecular docking verification using the AutoDockTools and AutoDock Vina tools and visual analysis using the PyMOL program were both completed.A total of 54active components,1 074 drug targets and 195 gene targets were obtained.There were 1 219 ONFH related targets.There were found to be39 pharmacological and illness interaction targets.The GO enrichment analysis produced 1 619 items,including 10 cellular components,1 517 biological processes,and 92 molecular activities,which is mainly related to angiogenesis,bone and lipid metabolism and inflammatory reaction.119 pathways were identified by the KEGG enrichment analysis,including the AGE-RAGE signaling route,the PI3K-Akt signaling pathway,and the IL-17 signaling pathway.Molecular docking results showed that quercetin,wogonin,and kaempferol active components had good affinity with IL6,p53 and VEGF core proteins.The high-frequency Chinese herbs of regulating marrow can treat ONFH by regulating immune inflammation,angiogenesis and bone metabolism.It has the characteristics of multi-component,multi-target and multi-path comprehensive treatment.