Combination of network pharmacology, in vitro experiments and molecular dynamic simulations revealed anti-inflammatory molecular mechanism of the shrub Metapanax delavayi

• Numerous bioactives were identified in extracts of Metapanax delavayi (MD). • Ninety-six of the identified compounds matched inflammatory targets. • MD extracts inhibited expression of several inflammation-related genes. • The MD metabolites formed stable complexes with phosphorylated proteins. The leaves of Metapanax delavayi shrub have been widely used in Chinese traditional diet to prepare tea-like beverages for alleviating inflammation. This study aimed to identify phytochemical components of M. delavayi leaf extract (MDE), and assess their anti-inflammation effects. The results showed that of the 101 compounds identified in the MDE, 96 were assessed as potentially anti-inflammatory based on network pharmacology analysis. The MAPK signaling pathway was surmised as the major anti-inflammatory pathway of action. The MDE enhanced the viability of the RAW264.7 cells, significantly inhibiting the overproduction of inflammatory factors (NO, IL-6, TNF-α, and PGE2), relative mRNA expression of TNF-α, iNOS, and COX-2, as well as several proteins (p-P38, p-ERK, p-JNK, iNOS, and COX-2) in lipopolysaccharide-induced RAW264.7 cells, meanwhile TGF-β1 production was increased. The key residues involved in binding to the top 10 leaf metabolites were Lys53, Thr179, Asp177, Glu147, and Pro174, in p-P38, p-ERK, p-JNK proteins. Molecular dynamic simulations demonstrated that predominantly three phosphorylated proteins formed stable and tightly bound complexes with certain metabolites of MDE.