A novel phosvitin phosphopeptide (Glu-Asp-Asp-pSer-pSer) as a potential food ingredient to intervene in osteoporosis: A potential mechanism study on promoting the osteogenic differentiation of MC3T3-E1 cells

Food-derived peptides with strong calcium-binding and osteogenic activity are promising functional ingredients for osteoporosis intervention. This study aims to explore whether a novel phosvitin phosphopeptide (Glu-Asp-Asp-pSer-pSer, EDDpSpS) with excellent calcium binding ability can promote the osteogenic differentiation of MC3T3-E1 cells and its potential mechanism. Network pharmacology analysis of EDDpSpS regulating osteoblast differentiation identified that the focal adhesion was a key signaling pathway, which is closely related to cell differentiation. Molecular simulations demonstrated that EDDpSpS was strongly bound to the target proteins within the focal adhesion signaling pathway and maintains a stable state. In addition, EDDpSpS had the potential to activate the key target protein Src tyrosine kinase (Src). Results of cell experiments showed that EDDpSpS (> 100 μg/mL) significantly promoted the differentiation and mineralization of MC3T3-E1 cells. Transcriptome analysis verified that the expression of paxillin (PXN), Src, actin beta/gamma 1 (ACTG1), and other osteoblast genes were regulated by EDDpSpS, leading to an up-regulation of the focal adhesion. In this study, the effect and the potential mechanism of EDDpSpS on osteoblast differentiation were clarified. EDDpSpS may be used as an effective dual-effect food ingredient for developing functional foods targeting osteoporosis. A novel phosvitin phosphopeptide (Glu-Asp-Asp-pSer-pSer) as a potential food ingredient to intervene in osteoporosis: a potential mechanism study on promoting the osteogenic differentiation of MC3T3-E1 cells. • Network pharmacology, transcriptomics, and molecular simulation were used to explore the mechanism of EDDpSpS promoting osteogenic differentiation. • EDDpSpS (> 100 μg/mL) significantly promoted the differentiation and mineralization of MC3T3-E1 cells. • EDDpSpS could effectively activate Src, up-regulated the focal adhesion signaling pathway, and ultimately promote osteoblast differentiation, thus intervening in osteoporosis. • EDDpSpS may be an effective dual-acting food ingredient for developing functional foods targeting osteoporosis.