Egg Yolk-Derived Peptide VPF Orchestrates OPG/RANKL/RANK-Mediated Signaling to Suppress Osteoclast Differentiation and Facilitate Bone Development in Zebrafish

High Resolution Image Download MS PowerPoint Slide This study was designed to comprehensively analyze the benefit effect and underlying mechanism of egg yolk-derived peptide VPF (Val-Pro-Phe) in bone development by integrating network pharmacology, molecular docking/dynamics simulation, and zebrafish in vivo experiments. Results demonstrated that VPF exhibited dose-dependent intervention effects within the concentration range of 0.2–10 μg/mL in normal and bone dysplasia zebrafish. Specifically, 0.2 μg/mL of VPF exhibited significant enhancements in total length (TL), mineralization, and swimming distance by 4.01%, 14.00% ( P < 0.05), and 7.31% in normal zebrafish model, whereas 1 μg/mL of VPF led to significant increases in the aforementioned indexes by 6.30%, 45.26%, and 51.05% ( P < 0.05) in bone dysplasia zebrafish model. Then, network pharmacology revealed that the highest correlation between VPF in improving bone development and osteoclast differentiation pathway, with osteoprotegerin (OPG) being the core node in this pathway. Molecular docking results showed that VPF exhibited the highest binding affinity for OPG, and this interaction was further validated by 200 ns molecular dynamics simulations, which demonstrated the stability of the VPF-OPG complex. Furthermore, VPF upregulated the gene expression of OPG, and downregulated the gene expression of the receptor activator of the NF-κB ligand (RANKL) and TNF receptor-associated factor 6 Gene (TRAF6), and downregulated the RANKL/OPG ratio in zebrafish. These data indicate that VPF alleviates skeletal dysplasia primarily by OPG-mediated suppression of osteoclast differentiation within the OPG/RANKL/RANK axis, thereby supporting its potential value as a functional food ingredient for promoting bone development.