Ameliorating effect of bovine bone collagen peptide/astragalus polysaccharide combination on bone mineral density and its underlying mechanism

Abstract

Bone mineral density (BMD) reduction leads to osteoporosis. Polypeptides and polysaccharides are each often used alone as supplements to improve BMD. However, the efficacy of their combination compared to single-component use, as well as the underlying synergistic mechanisms, remain unclear. Therefore, this study screened Bovine Bone Collagen Peptide (BBCP) and Astragalus Polysaccharide (APS) based on the theories of food-medicine homology and multi-target network regulation to investigate the synergistic ameliorative effects of the BBCP/APS combination (1:1, w/w) on BMD and its underlying mechanisms. An in vitro osteoblast model, in vivo ovariectomy-induced rat models, and network pharmacology analysis were employed to explore their combined effects on BMD and the associated mechanisms. In vitro, compared to either BBCP or APS alone, the BBCP/APS combination significantly enhanced MC3T3-E1 cell proliferation by 20.3 ± 5.80% and 22.9 ± 6.15%, respectively, and upregulated osteogenic markers, Alkaline phosphatase, Osteocalcin, and Type I collagen (compared with the model group, p < 0.01). In vivo, BBCP/APS (800 mg/kg) group increased the femoral BMD of ovariectomy-induced rats from 0.193 (model group) to 0.411 g/cm³ (p < 0.05) and improved trabecular bone microstructure. Concurrently, network pharmacology analysis identified ADRA1A and ADRA2A as core targets, and predicted the p38 MAPK pathway as the key signaling pathway. Activation of the p38 MAPK pathway by BBCP/APS was confirmed through RT-qPCR and Western blot. These findings indicated that the BBCP/APS combination could synergistically enhance BMD, overcoming the limitations associated with single-component use and providing valuable insights for the development of functional foods.