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Jiawei Duhuo Jisheng Mixture Attenuates H2O2-induced Oxidative Injury and Osteogenic Dysfunction in Primary
1Department of Orthopedic Surgery, The First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China.
Introduction:
Oxidative stress-induced osteoblast dysfunction is an important event in osteoporosis (OP). Jiawei Duhuo Jisheng Mixture (JDJM) is clinically used for bone-related disorders, but its molecular basis in OP remains unclear. This study explored OP-related candidate targets of JDJM and evaluated its cytoprotective effects in a hydrogen peroxide (H2O2)-induced primary osteoblast injury model.
Methods:
Candidate JDJM targets were predicted from its identified constituents and intersected with OP-related genes. The study integrated network-based and enrichment analyses with single-cell mapping, followed by molecular docking and molecular dynamics simulation. An H2O2-induced primary osteoblast injury model was used for experimental validation.
Results:
A total of 210 shared targets were identified. Network pharmacology and enrichment analyses implicated JDJM in the regulation of oxidative stress, inflammatory responses, extracellular matrix remodelling, osteogenesis and apoptosis, primarily through the MAPK, JAK-STAT and PI3K-Akt signalling pathways. Single-cell transcriptomic analysis further localized these candidate targets to osteogenic, stromal, vascular, and inflammatory cell populations. Molecular docking and dynamics simulations predicted stable interactions between representative JDJM-derived compounds and selected protein targets. In H2O2-injured primary osteoblasts, JDJM significantly reduced mean DCF fluorescence intensity, increased the alkaline phosphatase (ALP)-positive area and Alizarin Red S absorbance, and partially restored osteocalcin/bone Gla protein (OCN/BGP) protein and Bglap mRNA expression.JDJM was also associated with decreased total JAK1, STAT3, MAPK3, MMP9, and Bad expression and partial restoration of ESR1 expression.
Discussion:
JDJM attenuated H2O2-induced oxidative injury and osteogenic dysfunction in primary osteoblasts, accompanied by changes in JAK1, STAT3, MAPK3, MMP9, ESR1, and Bad expression. These results support a multi-target cytoprotective effect of JDJM under oxidative-stress conditions, although direct regulation of the corresponding signaling pathways remains to be confirmed.
Conclusion:
JDJM attenuated H2O2-induced oxidative injury and partially restored osteogenic function in primary osteoblasts, accompanied by coordinated expression changes in JAK1, STAT3, MAPK3, MMP9, ESR1, and Bad. These findings support a multi-target protective effect of JDJM against oxidative stress-related osteoblast dysfunction.