Hyperoside Modulates the Estrogen‐PI3K/VEGF Axis to Ameliorate Oxidative Damage‐Induced Inhibition of Bone Formation in MC3T3‐E1 Cells

OBJECTIVE: This study aims to investigate how hyperoside (HYP) alleviates oxidative stress-induced osteoporosis, its molecular mechanisms, and its impact on osteoblast differentiation, oxidative damage, and the estrogen-PI3K/VEGF signaling pathway. METHODS: The osteoblast differentiation model was induced using dexamethasone, and osteoblast-related markers like ALP, NO, GSH, MDA, and SOD were measured post-HYP intervention. A zebrafish model was used to assess HYP's impact on ROS and bone formation. Network pharmacology identified key oxidative stress and osteoporosis targets, with HYP's binding affinity confirmed via molecular docking and simulation. RT-qPCR verified the expression of key pathway targets. RESULTS: HYP can counteract dexamethasone-induced inhibition of osteoblast differentiation, boost ALP, NO, GSH, and SOD levels, and lower MDA levels in osteoblasts. It also reduces ROS accumulation and enhances bone formation in zebrafish. Network pharmacology identified a common oxidative stress and osteoporosis target, with molecular docking confirming HYP's stable binding. RT-qPCR showed HYP significantly upregulates SRC, PI3K, AKT1, and e-NOS, activating the estrogen-PI3K/VEGF pathway. CONCLUSION: HYP plays an anti-oxidative stress effect through targeted regulation of estrogen-PI3K/VEGF signal axis, and then promotes osteoblast differentiation and bone formation, which provides a new potential candidate drug and experimental basis for the treatment of osteoporosis.