基于网络药理学和分子对接技术探讨阳和汤治疗慢性骨髓炎的分子机制

Discussion on the mechanism of Yanghe Decoction in treating chronic osteomyelitis based on network pharmacology and molecular docking

目的利用网络药理学方法和分子对接技术研究阳和汤治疗慢性骨髓炎的活性成分和潜在作用机制。方法采用TCMSP、BATMAN-TCM数据库和相关文献筛选阳和汤组方药物活性成分和作用靶点,采用GeneCards、OMIM、DisGeNET、PharmGKB等数据库预测慢性骨髓炎的相关靶点。采用Cytoscape 3.8.0软件及STRING数据库构建“中药-活性成分-潜在靶点”网络、PPI网络,采用拓扑学参数筛选网络中核心成分及hub基因。采用MCODE插件对PPI网络进行蛋白模块聚类分析。采用KOBAS数据库对交集靶点进行GO功能和KEGG通路富集分析。采用AutoDock tool平台进行分子对接,验证主要活性成分与关键靶点的结合活性。结果获得阳和汤活性成分120个,作用靶点402个;慢性骨髓炎靶点1464个,阳和汤和慢性骨髓炎交集靶点103个,与交集靶点相关的活性成分110个,包括槲皮素、山柰酚等类黄酮化合物和β-谷甾醇、豆甾醇等植物甾醇;筛选出TNF、IL6、AKT1等9个hub基因,得到4个关键蛋白模块,涉及免疫炎症反应调控、血管微循环调节、骨的发育与形成、物质合成代谢等生理过程。通过富集分析得到193条信号通路,1552条GO条目。分子对接结果显示,阳和汤活性成分与关键靶点最低结合能均小于-5 kcal/mol,具有较好结合活性。结论阳和汤治疗慢性骨髓炎与多种成分、靶点、信号通路协同调控有关,主要通过TNF信号通路、IL-17信号通路、Toll样受体等通路调控TNF、IL6、CXCL8、VEGFA、AKT1等靶点,参与慢性骨髓炎局部的炎症反应、微循环、骨细胞代谢并干预致病菌的免疫逃逸机制。

Objective To study the active components and their potential mechanism of Yanghe Decoction for the treatment of chronic osteomyelitis(CO)via the methods of network pharmacology and molecular docking.Methods Active components and action targets of Yanghe Decoction were screened from TCMSP,BATMAN-TCM and relevant literature.GeneCards,OMIM,DisGeNET,and PharmGKB databases were used to predict the targets for the CO.Cytoscape 3.8.0 software and STRING database were used to build the networks of"Chinese materia medica-active components-potential targets"and"protein-protein interaction",and according to topological parameters in the network,the core active components as well as Hub genes were screened.MCODE plug was used to accomplish clustering analysis of protein modules in PPI network.Then,intersection targets were enriched and analyzed by GO and KEGG in KOBAS database.Finally,molecular docking was carried out with the help of Autodock tool platform to predict the binding ability between the main active components and key targets.Results A total of 120 active components of Yanghe Decoction and 402 targets were obtained;1464 CO-related targets were screened,and there were 103 intersection target genes of Yanghe Decoction-CO,110 active components related to intersection targets,which mainly contained some flavonoids and Phytosterols,such as quercetin,Kaempferol,and Beta-Sitosterol.There were 9 Hub genes,including TNF,IL6,AKT1,etc.,and 4 protein modules which involved the regulation of immune inflammatory response,vascular microcirculation,bone development,and formation,material synthesis and metabolism and other physiological processes.193 signaling pathways and 1552 GO results were acquired in KOBAS database.Molecular docking results showed that the active compounds had good binding activity with key targets based on the minimum binding energy of less than-5 kcal/mol.Conclusion The mechanism in the treatment of CO with Yanghe Decoction is a complex process of multiple components,multiple targets,and multiple pathways.It mainly regulates targets such as TNF,IL-6,CXCL8,VEGFA,and AKT1 through pathways such as TNF signaling pathway,IL-17 signaling pathway,and Toll-like receptors,participating in local inflammatory reactions,microcirculation,and bone cell metabolism in chronic osteomyelitis,and interfering with the immune escape mechanism of pathogenic bacteria.