Integrated Network Pharmacology Reveals the Mechanism of Action of Xianlinggubao Prescription for inflammation in Osteoarthritis

Abstract

Abstract Background. Osteoarthritis (OA) is a common chronic joint disease characterized by persistent inflammation. This study aimed to explore the potential mechanisms by which Xianlinggubao (XLGB) regulates OA inflammation via network pharmacology and in vitro experiments. Methods. Data on the bioactive compounds and targets of XLGB were obtained from the TCMSP, ETCM, SymMap, and ChEMBL databases. The targets of XLGB associated with OA inflammation were analyzed using differential expression analysis and OMIM, GeneCards, and PubMed Gene databases. Network pharmacology and bioinformatic analyses were performed to construct compound target and protein-protein interaction (PPI) networks to predict significant targets. Functional enrichment of the targets was performed using the ClusterProfiler package in R. Molecular docking, RT-PCR, and immunofluorescence assays were used to evaluate the ability of the compound to bind to anti-inflammatory targets during OA treatment. Results. Fifty-five bioactive compounds associated with 475 XLGB targets and 125 genes associated with OA inflammation were identified. The constructed PPI network revealed that XLGB may play therapeutic roles in OA inflammation by acting on the hub genes viz. monoclonal antibody to prostaglandin endoperoxide synthase 2, interleukin-1β, tumor necrosis factor, interleukin-6, and matrix metallopeptidase-9. Functional enrichment analysis suggested that XLGB exerted its pharmacological effects against OA inflammation by modulating multiple biological pathways, such as the IL-17, TNF, and NF-κB. The results of in vitro assays showed that XLGB can treat OA inflammation via hub genes, proteins, and signaling pathways. Conclusions: Using system pharmacology analysis, we systematically revealed the potential mechanisms of action of XLGB in OA inflammation, laying a foundation for its future clinical application.