<p>Chronic administration of glucocorticoids are being linked to an increased risk of osteoporosis. 5-Hydroxymethylfurfural (5-HMF) is a natural compound that possesses various biological activities, including osteogenic regulation. In this research, we sought to investigate the potential of 5-HMF to exert protective effects against glucocorticoid-induced osteoporosis. In cellular investigations, we deployed Western Blot (WB), Real-Time Quantitative real-time PCR (qRT-PCR), Alkaline phosphatase (ALP) and Alizarin S-red (ARS) staining to scrutinize the expression of osteogenic differentiation markers within MC3T3-E1 pre-osteoblasts and BMSCs. We used dexamethasone to establish a mouse model for glucocorticoid-Induced osteoporosis and administered 5-HMF to evaluate its influence on bone density and architecture through the use of micro-CT and histological analysis. In addition, we employed network pharmacology to elucidate the potential pathways and targets of 5-HMF. Finally, we explored the effect of AKT knockout on the treatment of 5-HMF. We found that 5-HMF significantly enhanced VEGFR2 phosphorylation, reestablishing angiogenesis and activating the PI3K/AKT pathway in vitro and in vivo. 5-HMF also inhibited Dexamethasone -induced apoptosis by regulating Bax and Bcl-2 expression. MK2206, an AKT inhibitor, abrogated 5-HMF’s protective properties against GCs. These results indicate that 5-HMF counteracts the negative effects of GCs on osteoblasts and has good bone-promoting differentiation effects.</p>