Network Pharmacology and Molecular Docking Combined with In Vivo Validation to Elucidate the Molecular Mechanisms of Adenophorae Radix in Fracture Healing

TL;DR

This study explores how Adenophorae Radix helps heal bone fractures by identifying its active compounds and their effects on bone regeneration processes.

Contribution

The study combines network pharmacology, molecular docking, and in vivo experiments to reveal the multi-target mechanisms of Adenophorae Radix in fracture healing.

Findings

Short-term AR treatment increased osteogenic markers RUNX2 and osteocalcin in bone marrow.

Long-term AR administration improved bone mineral density and callus maturation in fractured femurs.

Key compounds like cycloartenol acetate and β-sitosterol target HIF1A, PTGS2, and PPARG pathways involved in bone healing.

Abstract

Fracture healing is a multistage regenerative process requiring the coordinated regulation of inflammation, osteogenesis, and bone remodeling, yet pharmacological agents that effectively modulate these processes remain limited. Adenophorae Radix (AR), a traditional medicinal herb used for tissue repair, has not been mechanistically investigated in skeletal regeneration. In this study, a mouse femoral fracture model was employed to evaluate the effects of short-term (7 days) and long-term (5 weeks) oral administration of AR. Bone regeneration was assessed using micro-computed tomography, histological staining, and quantitative real-time polymerase chain reaction. Network pharmacology and molecular docking were applied to predict bioactive AR constituents and their target pathways, followed by in vivo validation. Short-term AR treatment significantly upregulated osteogenic markers, including RUNX2 and osteocalcin, in the bone marrow, indicating early activation of osteoblast differentiation. Long-term administration enhanced bone mineral density, trabecular organization, and callus maturation. Network pharmacology analysis identified cycloartenol acetate, β-sitosterol, and mandenol as major active compounds targeting osteogenesis- and osteoclast-related pathways, converging on HIF1A, PTGS2, and PPARG. Molecular docking demonstrated strong binding affinities between these compounds and their predicted targets, which was supported by increased expression of HIF1A, PTGS2, and PPARG in AR-treated femora. Collectively, these findings suggest that AR promotes fracture healing by regulating osteogenic differentiation and bone remodeling through multi-target transcriptional networks.

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