Integrated Network Pharmacology and Proteomics to Elucidate the Mechanism and Potential Targets of Tongfu Guanjie Gao on KOA in a Papain-Induced Rabbit Model

Objectives: Knee osteoarthritis (KOA) is a common chronic degenerative joint disease in the world. Tongfu Guanjie Gao (TFGJG) is external preparations of Traditional Chinese Medicine which has remarkable effect on relieving KOA. In this study, we aimed to construct a papain-induced KOA model in New Zealand white rabbits to investigate the mechanism of action of TFGJG in ameliorating KOA. Methods: After the successful induction of KOA rabbit model, TFGJG was given daily for 18 days for pharmacological intervention. The therapeutic effect of TFGJG on KOA was verified by H&E staining and ELISA assay, and the mechanism of action of TFGJG in improving KOA was explored by Network pharmacology and Proteomics. Results: Our results showed that TFGJG significantly improved Knee joint range of motion in KOA rabbits and reduced levels of tumor necrosis factor (TNF), interleukin-1beta (IL-1β), interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), collagen type II (COL-2) and matrix metalloproteinase-13 (MMP-13) inflammatory factors in synovial fluid. Network pharmacological analyses identified 218 targets of TFGJG for the treatment of KOA, and proteomics experiments showed that TFGJG regulates 150 differentially expressed proteins. Through comprehensive analysis, MAP2K1 and CRTAP were identified as the core targets of TFGJG in the treatment of KOA. Meanwhile, PRM verified that TFGJG significantly inhibited the expression of MAP2K1 and CRTAP in articular cartilage. Conclusion: TFGJG may be a promising agent that exerts the therapeutic effect on KOA by mediating inflammatory pathways, inhibiting the expression of MAP2K1 and CRTAP and offers potential new drug targets for the treatment of KOA.