OBJECTIVE To identify the chemical constituents of the Lithospermum zollingeri DC.-Prunella vulgaris L. herb pair, screen its active components using network pharmacology, and explore the material basis of its anti-inflammatory efficacy through cellular experimental verification.

METHODS UHPLC-Q-TOF-MS was employed to identify the chemical constituents of Lithospermum zollingeri and Prunella vulgaris. Active components were screened via the Swiss ADME platform, SwissTargetPrediction, GeneCards, and OMIM databases. The STRING database and Cytoscape 3.10.0 were used to construct a protein-protein interaction(PPI) network. GO functional annotation and KEGG pathway enrichment analyses were performed using the DAVID database. A lipopolysaccharide(LPS)-induced inflammatory model in RAW264.7 macrophages was established. The contents of COX-2, PGE2, and MCP-1 in cell supernatants, as well as COX-1 in cell lysates, were detected by ELISA to verify the anti-inflammatory effects of extracts and active components and explore their pharmacodynamic material basis.

RESULTS A total of 56 compounds were identified, among which the UHPLC-Q-TOF-MS characterization of Lithospermum zollingeri was reported for the first time. Network pharmacology screening identified morin, esculetin, and caffeic acid as key active components; core targets included GAPDH, TNF, ALB, AKT1, EGFR, etc. GO enrichment analysis revealed targets’ involvement in processes such as response to external stimuli, positive regulation of cell migration, and inflammatory response, etc. KEGG enrichment analysis revealed significant enrichment of signal pathways in cellular response to nitrogen compounds, inflammatory response and cancer pathways. Cellular experiments showed that extracts of Lithospermum zollingeri, Prunella vulgaris, their herb pair, and the monomer compounds(morin, esculetin, caffeic acid) significantly inhibited LPS-induced production of MCP-1, COX-2, and PGE2, but had no significant effect on the protein expression of COX-1.

CONCLUSION The Lithospermum zollingeri-Prunella vulgaris herb pair exerts its anti-inflammatory effects through a multi-component, multi-target, multi-pathway synergistic mechanism. This study provides a theoretical foundation for elucidating the pharmacodynamic material basis underlying its anti-inflammatory efficacy.