Osteoarthritis (OA) is a prevalent chronic joints disorder, characterized by progressive degeneration of articular cartilage accompanied by joint inflammation. The development of a safe and effective pharmacological therapy is highly anticipated. This research was designed to comprehensively investigate the function of NEF in regulating chondrocyte autophagy, along with its influence on the advancement of OA. In this study, a destabilization of the medial meniscus (DMM) surgery-induced osteoarthritis (OA) rat model and a tert-butyl hydroperoxide (TBHP)-stimulated primary rat chondrocyte model were established. By integrating network pharmacology, gene knockout, flow cytometry, and Western blotting analyses, the protective effects of neferine (NEF) against OA and its possible underlying molecular mechanisms were investigated. NEF effectively mitigates TBHP-induced chondrocyte apoptosis and senescence phenotypes while activating autophagy. Molecular docking, molecular dynamics simulations, and gene knockdown experiments collectively suggest that NEF may enhance autophagy fluxes by inhibiting the downstream mTOR signaling pathway, thereby exerting partial protective effects against osteoarthritis. Furthermore, in a rat osteoarthritis model established through medial meniscus destabilization, NEF treatment greatly reduced the degeneration of articular cartilage caused by surgery. Collectively, these results indicate that NEF may enhance autophagy fluxes by inhibiting the mTOR signaling pathway, thereby exerting partial ameliorative effects on OA progression and highlighting its therapeutic potential for OA intervention.
