Asarum heterotropoides F. schmidt attenuates osteoarthritis via multi-target anti-inflammatory actions: A network pharmacology and experimental validation

Abstract

Ethnopharmacological relevance: Asarum heterotropoides F. Schmidt (ARR) has a well-documented history of traditional use in East Asia for musculoskeletal pain disorders, including osteoarthritis (OA), attributed to its significant anti-inflammatory properties. While preliminary studies suggest potential anti-inflammatory effects, conclusive evidence regarding the ability of ARRs to modulate the multiple inflammatory pathologies involved in OA pathogenesis is currently lacking.

Aim of the study: This study aimed to experimentally evaluate the effects of ARR extract on pain, cartilage integrity, and inflammatory responses using in vitro and in vivo models relevant to OA, guided by initial computational predictions.

Materials and methods: Active ingredients of ARRs were retrieved from four databases and screened using SwissADME for ADME predictions. Disease targets were combined with OA-related genes from GEO microarray database. The intersecting genes underwent protein-protein interaction construction, GO, and KEGG enrichment analysis. A compound-target-pathway network was constructed using Cytoscape and was validated via molecular docking. Pain-relieving, functional, and chondroprotective effects were assessed in vivo using acetic acid-induced peripheral pain mice and monosodium iodoacetate (MIA)-induced osteoarthritis rat models. Furthermore, anti-inflammatory properties were explored by evaluating serum cartilage tissue and lipopolysaccharide-stimulated RAW 264.7 cells.

Results: Network pharmacology analysis elucidated five principal active constituents of ARR (cryptopine, 5-[2-(2-hydroxyphenyl)ethyl]-2,3-dimethoxy-phenol, 5-[2-(3-hydroxyphenyl)ethyl]-2-methoxybenzene-1,3-diol, naringenin, resorstatin) alongside 22 putative herbal targets. Molecular docking analyses revealed strong binding affinities (-8 to -9.4 kcal/mol) of these constituents towards principal target proteins. Functional GO and KEGG enrichment analyses indicated that ARR exerts its effects potentially involving pathways associated with cancer, fluid shear stress, and atherosclerosis. In vivo assessments demonstrated significant mitigation of pain, functional deficits, and cartilage degradation by ARR within an MIA-induced osteoarthritis model. Molecular dynamics simulations validated stable interactions between the primary compounds and their designated target proteins. The therapeutic efficacy of ARR was characterized by dose-dependent suppression of diverse inflammatory mediators (IL-1β, IL-6, TNF-α), matrix metalloproteinases (MMP-1, -3, -8, -13), and signaling pathways including CCND1, CDK2, IKBKB, HIF1A, BDKRB1, SIRT1, MAPK8, and NLRP3 within both RAW264.7 cells and articular cartilage tissue.

Conclusions: This investigation demonstrates that ARR exerts pain alleviation, functional enhancement, and chondroprotective effects in osteoarthritis via multi-target anti-inflammatory actions. Integrating network pharmacology, molecular docking, animal models, and cellular experiments, this study comprehensively elucidated the multifaceted anti-inflammatory mechanisms attributed to ARR. These findings collectively provide a crucial foundation for understanding the potential therapeutic efficacy and operative mechanisms of ARR for osteoarthritis management.

Keywords: Asarum heterotropoides F.Schmidt; GEO datasets; Molecular docking; Network pharmacology; Osteoarthritis.