Network Pharmacology and In Vivo Validation Reveal the Potential Protective Mechanisms of Chonggu Granules in Knee Osteoarthritis
Yuanyuan Cheng, Yawen Zhu, Ai Qian, Juan Zhang, Chuanbing HuangIntroduction:
Knee osteoarthritis (KOA) is a common degenerative joint disorder characterized by progressive cartilage loss, chronic low-grade inflammation, and oxidative injury. Ferroptosis-associated iron accumulation and lipid peroxidation have been implicated in osteoarthritic cartilage damage. This study investigated the protective effects and potential multi-target mechanisms of Chonggu Granules (CGG) in KOA.
Methods:
Network pharmacology and molecular docking were used to identify candidate CGG compounds, targets, and pathways. KOA was induced in rats by intra-articular papain injection. Rats were randomly allocated to the Control, Model, glucosamine hydrochloride (AT), CGG, Ferrostatin-1 (Fer-1), and CGG plus ML385 groups. Cartilage histopathology, mitochondrial ultrastructure, chondrocyte apoptosis, oxidative stress, serum Fe2+ levels, lipid peroxidation, inflammatory cytokines, and the expression of Nrf2 and related antioxidant and ferroptosis-associated molecules were evaluated.
Results:
Network pharmacology identified 115 overlapping CGG-KOA targets, including the hub target NFE2L2, enriched in pathways related to inflammation, oxidative stress, and ferroptosis. Molecular docking predicted favorable binding between several major CGG compounds and multiple hub targets. In vivo, CGG attenuated cartilage degeneration, mitochondrial abnormalities, chondrocyte apoptosis, inflammatory cytokine production, oxidative stress, elevated serum Fe2+ levels, and lipid peroxidation. CGG also partially normalized the expression of Nrf2, HO-1, GPX4, and other ferroptosis-associated molecules. These effects were partially attenuated by ML385.
Discussion:
The findings suggest that CGG exerts multi-component and multi-target protective effects against KOA-related cartilage injury by modulating inflammation, oxidative stress, and ferroptosis-associated processes. The partial attenuation of these effects by ML385 supports the involvement of Nrf2-associated antioxidant signaling and downstream ferroptosis-related responses.
Conclusion:
CGG alleviated KOA-related cartilage injury and ferroptosis-associated oxidative damage through potential multi-target regulation, with Nrf2-associated antioxidant responses contributing to these protective effects.