Integrating novel network pharmacology and experimental validation to study the mechanism of Wutou decoction against rheumatoid arthritis

Wutou decoction (WTD) is a traditional remedy in Chinese medicine for rheumatoid arthritis (RA). However, its active ingredients are complex, and how it counteracts RA is still not clear. The objective of this research was to investigate the key components and targets of WTD action. RA-related genes were analyzed using the GSE55235 dataset. The active components of WTD were analyzed using online databases. Component-target-pathology (CTP) networks were built using the Cytoscape software. The functions and pathways of the key targets were analyzed using GO and KEGG. The component-target interaction was assessed by molecular docking. Moreover, the impact of WTD on RA progression and its mechanism were validated using cell and animal experiments. The results showed that 1361 genes were related to RA, and they were enriched in 2035 biological processes and 97 signaling pathways. The five herbs of WTD had 221 common targets, and each herb had numerous components. A total of 66297 interactions were found among 440 active ingredients and 2638 targets within the WTD in the CTP network. Next, 114 core functional component groups were selected, and the types and quantities of target-enriched signaling pathways they covered were greater than those of RA-related genes alone. In vivo, WTD attenuated RA-induced arthritis and bone destruction. In vitro, benzoic acid, quercetin, apigenin, and decanoic acid inhibited the inflammatory response of macrophages by inactivating the PI3K/ATK and MAPK pathways. In conclusion, WTD exerts a therapeutic effect on RA through multi-component and multi-target, providing novel methods to study the mechanism of WTD.