Pinoresinol diglucoside activates the Wnt/beta-catenin signaling pathway to protect osteoblasts

doi:10.12307/2025.237

Abstract

Abstract: BACKGROUND: Pinoresinol diglucoside promotes bone formation and bone matrix synthesis and accelerates bone tissue repair. However, the mechanism of action and effects of this compound in osteoblasts need to be further explored.
OBJECTIVE: To investigate the effect and mechanism of action of pinoresinol diglucoside on dexamethasone-treated osteoblasts based on the Wnt/β-catenin signaling pathway.
METHODS: Different concentrations of dexamethasone groups and pinoresinol diglucoside groups were set to treat osteoblasts for 24 hours, and the optimal intervention concentrations were screened. Osteoblasts were treated with dexamethasone, pinoresinol diglucoside and inhibitor XAV-939. Then, control group, dexamethasone group, XVA-939 group, pinoresinol diglucoside group, pinoresinol diglucoside+XVA-939 group were set up. Cell counting kit-8 assay was used to detect cell activity. Alkaline phosphatase activity and caspase3/7 enzyme activity in cells were detected. Annexin V/PI staining and EdU assay were used to detect cell apoptosis and proliferation. Real-time qPCR and western blot were used to detect the mRNA and protein expression levels of Wnt3a, β-catenin, c-myc, osteocalcin, and type I collagen, respectively.
RESULTS AND CONCLUSION: After dexamethasone and pinoresinol diglucoside intervened in osteoblasts for 24 hours, 10 μmol/L dexamethasone was found to be the optimal intervention concentration for cell inhibition, and cell proliferation was most pronounced at a concentration of pinoresinol diglucoside of 100 μmol/L. Compared with the dexamethasone group, alkaline phosphatase activity was significantly enhanced (P < 0.05) and caspase3/7 enzyme activity was significantly reduced (P < 0.05) in the pinoresinol diglucoside group. Annexin V/PI staining and cell proliferation assay by EdU method showed that pinoresinol diglucoside inhibited apoptosis and promoted proliferation of osteoblasts after dexamethasone intervention. The mRNA and protein expression levels of Wnt3a, β-catenin, c-myc, osteocalcin, and type I collagen were significantly higher in the pinoresinol diglucoside group and pinoresinol diglucoside+XVA-939 group compared with the dexamethasone and XVA-939 groups (P < 0.05). To conclude, pinoresinol diglucoside can inhibit osteoblast apoptosis after dexamethasone intervention, protect osteoblast activity and promote osteoblast proliferation by activating the Wnt/β-catenin signaling pathway, which may play a role in delaying steroid-induced osteonecrosis of the femoral head .

Key words: pinoresinol diglucoside, steroid-induced osteonecrosis of the femoral head, osteoblast, cell activity, Wnt/β-catenin signaling pathway, cell proliferation, cell apoptosis

CLC Number:

Yu Peng, Meng Dongfang, Li Huiying, Liu Hongfei, He Zike. Pinoresinol diglucoside activates the Wnt/beta-catenin signaling pathway to protect osteoblasts [J]. Chinese Journal of Tissue Engineering Research, 2025, 29(2): 339-346.

Figures/Tables 11

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