Abstract

Ethnopharmacological relevance Mulberry ( Morus alba L.) has a longstanding history of use in Traditional Chinese Medicine (TCM). It is recorded in the classic Compendium of Materia Medica (Ben Cao Gang Mu) that mulberry possesses the efficacy to “strengthen muscles and bones”. This traditional indication suggests a potential role in skeletal health, yet its scientific and pharmacological basis has not been fully elucidated. The present study provides the first experimental evidence that a standardized polyphenol extract from mulberry (ABRU) bidirectionally regulates bone metabolism, simultaneously promoting bone formation and inhibiting bone loss. This work not only validates a centuries-old ethnopharmacological claim with modern molecular and functional evidence but also identifies mulberry polyphenols as a promising candidate for developing natural product-based therapies for bone metabolic disorders like osteoporosis. Aim of the study This study aims to investigate the dual regulatory effects and mechanisms of mulberry polyphenols (ABRU) on bone metabolism. Methods MC3T3 cells and Bone marrow-derived macrophages (BMDMs) were used to evaluate the effects of various mulberry extracts on osteoblastic mineralization and osteoclastogenesis. Active extracts were analyzed for their chemical composition using liquid chromatography-mass spectrometry (LC-MS). Network pharmacology, molecular docking, ROC analysis, and thermal shift assays were employed to identify the molecular targets of ABRU. The dual regulation of bone metabolism by ABRU was further assessed in vivo using subcutaneous bone formation assays and a D-galactose-induced mouse model of bone loss. Results In vitro experiments demonstrated that mulberry polyphenol (ABRU) significantly enhanced osteoblast proliferation and differentiation. After 24 h, osteoblast proliferation doubled, after 14 days, mineralization (measured by Alizarin Red staining) and alkaline phosphatase activity both increased two-fol