Abstract
Steroid-induced osteonecrosis of the femoral head (SONFH) is a severe bone disease associated with long-term glucocorticoid use, characterized by impaired bone metabolism and vascular insufficiency. Bilobalide (BB), a natural sesquiterpene from Ginkgo biloba, exhibits anti-apoptotic, antioxidant, and pro-angiogenic properties, yet its role in SONFH remains unclear. We integrated network pharmacology and molecular docking to predict the targets and pathways of BB in SONFH. Key targets were validated using molecular docking software. For in vivo experiments, a rat SONFH model was established using methylprednisolone (MPS), and BB was administered orally. Micro-CT, H&E staining, TUNEL assay, and immunohistochemistry were employed to evaluate bone microstructure, apoptosis, and the expression of osteogenic and angiogenic markers. Immunofluorescence was used to assess HIF-1α expression in rat femoral head tissues. For in vitro experiments, MC3T3-E1 osteoblasts were treated with dexamethasone(DEX) and BB. Cell viability was detected using the CCK-8 assay, and the protein levels of the HIF-1α and ERK pathways were examined by Western blot. Network pharmacology identified 94 common targets between BB and SONFH, with enrichment in HIF-1 and ERK signaling pathways. Molecular docking confirmed strong binding affinities between BB and core targets. In MPS-induced rats, BB treatment significantly improved bone mineral density, trabecular microstructure, and reduced osteocyte apoptosis. BB also upregulated HIF-1α, Runx2, OCN, CD31, and VEGF expression, indicating enhanced osteogenesis and angiogenesis. In vitro, BB rescued dexamethasone-induced suppression of osteoblast viability and upregulated the ERK/HIF-1α pathway. Bilobalide attenuates SONFH progression by activating the ERK/HIF-1α signaling pathway, promoting osteogenesis and angiogenesis, and reducing osteocyte apoptosis. These findings highlight BB as a promising candidate for SONFH prevention and support the utility of network pharmacology in mechanistic natural product research.
Data availability
The authors affirm that all data supporting the findings are completely accessible and can be acquired by submitting a request to the corresponding author.
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Funding
This study was funded by the Clinical Collaboration and Innovation Project of Integrated Traditional Chinese and Western Medicine by the Shaanxi Provincial Administration of Traditional Chinese Medicine (No.2020ZXY010) (to Yangquan Hao).
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Conceptualization, Qi Chen and Yangquan Hao; Data curation, Qi Chen and Yangquan Hao; Formal analysis, Qi Chen, Bo Wang, Hu Liang, and Hanbo Xu; Funding acquisition, Yangquan Hao; Investigation, Qi Chen, Bo Wang, Hanbo Xu, Hu Liang, and Kun Zhang; Methodology, Qi Chen; Project administration, Yangquan Hao; Resources, Yangquan Hao; Software, Qi Chen, Bo Wang and Yangquan Hao; Supervision, Kun Zhang and Yangquan Hao; Validation, Qi Chen, Bo Wang, Hu Liang and Hanbo Xu; Visualization, Qi Chen and Bo Wang; Writing – original draft, Qi Chen and Bo Wang; Writing – review & editing, Yangquan Hao. All authors have read and agreed to the published version of the manuscript.
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All animal experiments were conducted in accordance with the ARRIVE guidelines 2.0. The study received approval from the Ethics Committee of the Institutional Animal Care and Use Committee at Shaanxi University of Chinese Medicine (Approval number SUCMDL20241124001, approved on 24 November 2024).
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The study did not involve humans.
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This manuscript includes results and/or pathway maps generated from the KEGG database. Formal permission to publish this material under a CC BY license has been obtained from Kanehisa Laboratories.
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Chen, Q., Wang, B., Liang, H. et al. Bilobalide attenuates steroid-induced osteonecrosis of the femoral head by upregulating the ERK/HIF-1α signaling pathway and promoting angiogenesis-osteogenesis coupling. Sci Rep (2026). https://doi.org/10.1038/s41598-026-37128-3
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DOI: https://doi.org/10.1038/s41598-026-37128-3
