基于网络药理学和试验验证探讨镰形棘豆黄酮类化合物对于炎症的作用机制

Study the mechanisms of flavonoids from Oxytropis falcata bunge on inflammation by network pharmacology combined with experimental verification

目的:利用网络药理学和分子对接方法研究镰形棘豆黄酮类化合物(Flavonoids of Oxytropis falcata Bunge,FOFB)抗炎的分子机制。方法:通过SEAdatabase、SwissTarget Prediction和TargetNet数据库预测TFOFB相关靶点。从DisGeNET、GeneCards和Geneclip3数据库收集与炎症相关的靶点,通过靶点映射确定活性化合物作用于炎症的靶点。利用Cytoscape 3.7.2软件构建药物-化合物-疾病-靶点蛋白互作网络图。借助Omicshare和Metascape数据库分别对化合物与炎症的交集靶点进行GO及KEGG富集分析。用Griess法检测活性化合物对脂多糖(LPS)诱导的RAW264.7巨噬细胞iNOS的影响,以验证其抗炎活性。通过Autodock vina软件对核心化合物和核心靶点进行分子对接验证。结果:镰形棘豆黄酮类化合物与炎症共同靶点432个,关键靶点5个,包括AKT1、HSP90AA1、EP300等。KEGG富集结果显示FOFB抗炎主要调控cAMP信号通路、NF-κB信号通路、cGMP-PKG信号通路、Trp通道等。分子对接显示核心成分与核心靶点对接良好。7-羟基黄烷酮、2’-4’-二羟基查尔酮这两种化合物可显著抑制LPS诱导RAW264.7巨噬细胞释放i NOS,具有明显的抗炎作用。结论:镰形棘黄酮类化合物通过“多成分-多靶点-多通路”发挥抗炎作用,该作用可能与对含氧化合物的反应、对内源性刺激的反应、对氮化合物的反应等生物学过程有关,可为未来深入研究镰形棘豆黄酮类化合物奠定理论基础。

Objective:To study anti-inflammatory molecular mechanism of Flavonoids of Oxytropis falcata Bunge(FOFB)using network pharmacology and molecular docking methods.Methods:TFOFB-related targets were predicted through SEA database,SwissTarget Prediction and TargetNet database.Inflammation-related targets were collected from DisGeNET,GeneCards and Geneclip3 databases.Targets of active compounds on inflammation were determined through target mapping.Cytoscape 3.7.2 software was used to construct a drug-compound-diseasetarget protein interaction network diagram.With the help of Omicshare and Metascape databases,GO and KEGG enrichment analysis were performed on intersection target of the compound and inflammation.The Griess method was used to detect effect of active compounds on iNOS of RAW264.7 macrophages induced by lipopolysaccharide and to verify its anti-inflammatory activity.Autodock vina software is used to verify molecular docking of core components and important targets.Results:There were 432 common targets of Oxytropis falciparum flavonoids and inflammation,and 5 key targets,including AKT1,HSP90AA1,EP300,etc.KEGG enrichment results show that FOFB antiinflammatory mainly regulates cAMP signaling pathway,NF-κB signaling pathway,cGMP-PKG signaling pathway,Trp channel and so on.Molecular docking shows that core component is well docked with core target.The two compounds of 7-hydroxyflavanone,and 2’-4’-dihydroxychalcone can significantly inhibit the release of iNOS from RAW264.7 macrophages induced by LPS,and have obvious anti-inflammatory effects.Conclusion:Flavonoids of Oxytropis falcata Bunge exert an anti-inflammatory effect through“multi-component-multi-target-multipathway”,which may relate to the responses to oxygen-containing compounds,endogenous stimuli,and nitrogen compounds,etc.The research can lay a theoretical foundation for in-depth study of Flavonoids of Oxytropis falcata Bunge in the future.