Anti-inflammatory properties of Fangji Huangqi tang: Novel application based on integrated network pharmacology combined with in vitro validation

Fangji Huangqi Tang (FHT) is a classic traditional Chinese medicine (TCM) formula widely used in clinical practice. In this paper, a novel application of FHT for inflammation has been identified by network pharmacology-based computational methods and in vitro validation. The bioactive compounds of FHT were screened from the TCMSP and BATMAN-TCM databases. Then, the biological targets related to the chemical components and inflammation were obtained from the TCMSP, BATMAN-TCM, TTD, OMIM and Uniprot databases. Subsequently, an integrated ‘compound-target-disease’ network and enrichment analysis were constructed via Cytoscape and R software. Molecular docking and an LPS-induced inflammation model were further used to identify the binding ability and the effect of active constituents on inflammation-related targets. Eventually, a non-interactive literature-based discovery (NILBD) was conducted by the Arrowsmith online tool for the deep validation of the pharmacological mechanism of key components contained in FHT. A total of 17 bioactive compounds, including 42 potential targets of FHT and 205 related targets involved in inflammation, were subjected to network analysis. Thirteen intersection targets indicated the principal elements linked to inflammation therapy. Gene Ontology (GO) analysis was performed, and 7 related signaling pathways were revealed by the Kyoto Encyclopedia of Genes and Genomes (KEGG). Subsequently, the Calycosin-PTGS2 axis with tight binding affinity (BA) as well as anti-inflammatory activity was manifested by molecular docking and an in vitro experiment as the critical compound-target link. Meaningful relations between Calycosin and inflammation were implied through Arrowsmith, which overlapped with the findings of enrichment analysis, revealing that MAPK and NF-κB could be regulators of PTGS2. The present study explored the potential targets and signaling pathways of FHT against inflammation, which may help to illustrate the mechanisms of FHT and provide a better understanding of its anti-inflammatory effects.