Integrative Network Pharmacology, Molecular Docking, and In vitro Investigation of Mentha Arvensis L. Leaf Extract as a Multi-Target Modulator of Rheumatoid Arthritis Pathways

Objective: Rheumatoid arthritis (RA), a chronic inflammatory disease, may cause multi-joint effects that severely impair quality of life. This study evaluated the molecular docking, in vitro validation and network pharmacology potential of the ethanolic leaf extract of Mentha arvensis L. in targeting RA-associated pathways. Methods: Shade-dried and ground M. arvensis leaves were subjected to a five-day cold maceration process in 80% ethanol. Bioactive substances were screened for drug-likeness and evaluated for their potential RA targets. Network pharmacology was employed to identify key genes and signalling pathways, and molecular docking was used to study ligand-target interactions. The extract was tested in vitro for its antioxidant and anti arthritic properties. Results: Seven out of 101 phytoconstituents fulfilled drug-likeness criteria and were selected for further study. Network analysis revealed that PTGS1/2 (prostaglandin-endoperoxide synthase 1/2), PRKCQ (protein kinase C theta), SYK (spleen-associated tyrosine kinase), PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha), and PIK3CD (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit delta) were key targets. Pathway enrichment identified MAPK (mitogen-activated protein kinase), PI3K-AKT, TNF (tumor necrosis factor), and arachidonic acid metabolism pathways as primary mediators. Kaempferol glucoside demonstrated the highest binding affinities (docking scores: −7.2 to −9.4 kcal/mol). The extract showed significant antioxidant activity with an IC₅₀ (half-maximal inhibitory concentration) of 54.24 µg/ml for 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and an IC₅₀ of 209.93 µg/ml for bovine serum albumin (BSA) heat-denaturation inhibition. Conclusion: The findings indicate that the anti-arthritic effects of Mentha arvensis are mediated through a complex network of molecular targets and pathways. These results provide preliminary evidence warranting further preclinical and clinical studies to evaluate its utility as a natural therapeutic agent for RA.