Study on the regulation and mechanism of kaempferol-3-O-arabinoside on osteogenic differentiation of bone marrow mesenchymal stem cells

doi:10.19438/j.cjoms.2025.05.002

Abstract

Abstract: PURPOSE: To study the effect of kaempferol-3-O-arabinoside(KA) on osteogenic differentiation and epidermal growth factor receptor(EGFR), mitogen-activated protein kinase(MAPK) pathway by stimulating bone marrow mesenchymal stem cells (BMSCs). METHODS: BMSCs were isolated from the extraction sockets of impacted teeth in healthy young males, cultured until passage 3, and then treated with different concentrations of KA. CCK-8 assay was used to determine cell proliferation activity. Alkaline phosphatase (ALP) and alizarin red S (ARS) staining were employed to detect the changes in osteogenic capacity of BMSCs after KA treatment, and ImageJ software was used to analyze ALP activity. Real-time quantitative reverse transcription PCR (RT-qPCR) and Western blot were used to detect the changes in the gene and protein expressions of osteogenic differentiation-related genes including Runt-related transcription factor 2 (RUNX2), type I collagen (COL1), and osteopontin (OPN). Network pharmacology was adopted to analyze the main action targets and potential related signaling pathways of KA. BMSCs were treated with 5 μmol/L EGFR inhibitor gefitinib, 10 μmol/L MEK1/2 inhibitor U0126, and 10 μmol/L p38 inhibitor SB203580 to inhibit the main signaling molecules of the MAPK signaling pathway, and then RT-qPCR and Western blot were used to detect the changes in the osteogenic capacity of BMSCs. RESULTS: The results of CCK-8 showed that 80 and 160 μmol/L KA significantly affected the activity of BMSCs, and 10, 20 and 40 μmol/L KA were used for further experiments. The results of ALP and ARS staining indicated that 10 μmol/L KA had the best osteogenic induction effect. The results of RT-qPCR and Western blot showed that KA at 10 μmol/L had the best effect on the expression of mRNA and protein related to osteogenesis. The results of prediction analysis suggested that KA might promote osteogenic differentiation of BMSCs through EGFR- MAPK pathway. Western blot results showed that BMSCs activated MAPK signaling pathway after KA treatment, and the osteogenic promotion effect of BMSCs was decreased after treatment with MAPK signaling pathway-related inhibitors. CONCLUSIONS: Certain concentration of KA can stimulate and regulate BMSCs, activate EGFR-MAPK-RUNX2 signaling pathway and promote osteogenic differentiation.

Key words: Kaempferol-3-O-arabinoside, Bone marrow mesenchymal stem cells, Osteoblast, MAPK signaling pathway, Epidermal growth factor receptor, Osteogenic differentiation

CLC Number:

R782
Q254

Zhang Haoze, Ren Haining, Zhu Qinghai, Wang Chenxing, Ye Jinhai. Study on the regulation and mechanism of kaempferol-3-O-arabinoside on osteogenic differentiation of bone marrow mesenchymal stem cells[J]. China Journal of Oral and Maxillofacial Surgery, 2025, 23(5): 434-441.

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