Screening the anti-neuroinflammatory active fraction of Wenmian San and exploring the underlying mechanism by UPLC–Triple-TOF-MS/MS-based network pharmacology, DEGs, WGCNA, and molecular docking

This study aimed to screen the anti-neuroinflammatory active fraction of Wenmian San and to elucidate its underlying mechanisms. A lipopolysaccharide (LPS)-induced BV2 cell model was established to evaluate the anti-neuroinflammatory activities of different fractions; the ethyl acetate fraction (EAF) exhibited the most potent inhibitory effects on NO production and cytokine release. Chemical analysis identified 41 compounds, mainly including flavonoids, coumarins, and phenylpropanoids. Network pharmacology integrated with weighted gene co-expression network analysis (WGCNA) and differentially expressed genes (DEGs) analysis identified ICAM1 and NFKB1 as core target genes, with isorhamnetin, quercetin, byakangelicin, kaempferol, imperatorin, and isoliquiritigenin as the major active components. Molecular docking analysis further verified favourable binding affinity between the screened active compounds and the identified targets. Overall, the EAF of Wenmian San exerts significant anti-neuroinflammatory effects through a multi-component, multi-pathway, and multi-target therapeutic pattern.