Serum metabolomics-driven network pharmacology elucidate the anti-rheumatoid arthritis potential of garden cress.

Garden cress (Lepidium sativum L.) has been traditionally utilized for the treatment of various diseases and is increasingly consumed as a functional food and alternative medicine in many countries due to its therapeutic potential. Notably, L. sativum is a promising candidate for mitigating rheumatoid arthritis (RA). This study employed a serum pharmacochemistry approach combined with a network pharmacology strategy to identify the active components and elucidate the underlying mechanisms of L. sativum in RA management. An RA rat model was established using Complete Freund's Adjuvant (CFA). Following L. sativum administration, bioactive serum components were identified and quantified as markers of its pharmacological activity. Twenty-six serum metabolites, including 11 prototype compounds and 15 derived metabolites, were identified as key bioactive constituents absorbed at significant concentrations, potentially mediating the anti-RA effects of L. sativum. Among these, fatty acids and their conjugated metabolites emerged as the most relevant. Through network pharmacology, potential target genes and associated pathways were predicted. KEGG pathway analysis highlighted critical RA-related pathways, including arachidonic acid metabolism, modulation of inflammatory regulators in TRP channels, linoleic acid metabolism, and antifolate resistance pathways. Experimental data demonstrated that L. sativum significantly downregulated key inflammatory mediators such as IL-1β, TNF-α, MMP-9, CYP1A2, PLA2G2A, and MAPK8. This integrated study provides insight into the molecular mechanisms and active constituents of L. sativum, serving as a foundational reference for its therapeutic application against RA.