咖啡通过抑制Wnt/β-catenin通路介导大鼠骨髓间充质干细胞成骨分化

doi:10.13591/j.cnki.kqyx.2026.05.003

摘要/Abstract

摘要：

目的探究咖啡对大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMMSCs）成骨分化的影响，并初步阐明其与Wnt/β-catenin信号通路的关系。方法通过全骨髓贴壁法分离并培养大鼠BMMSCs，制作不同浓度咖啡条件培养基，碱性磷酸酶染色筛选最适作用浓度，通过qRT-PCR和Western blot观察成骨相关指标变化，并检测β-catenin基因和蛋白的表达水平变化。结果成功分离并培养出大鼠BMMSCs。ALP活性检测筛选出0.1 mg/mL为咖啡条件培养基最适作用浓度，进一步实验发现，其在前3 d对大鼠BMMSCs增殖无显著影响，在5~9 d时抑制大鼠BMMSCs增殖。咖啡条件培养基抑制大鼠BMMSCs成骨指标的表达，同时抑制Wnt/β-catenin信号通路有关蛋白的表达。结论咖啡条件培养基可抑制大鼠BMMSCs成骨分化，其作用机制可能与抑制Wnt/β-catenin信号通路有关。

关键词: 咖啡, 骨髓间充质干细胞, 成骨分化, Wnt/β-catenin信号通路

Abstract:

Objective To investigate the effect of coffee on the osteogenic differentiation of rat bone marrow mesenchymal stem cells （BMMSCs） and preliminarily elucidate its relationship with the Wnt/β-catenin signaling pathway. Methods Rat BMMSCs were isolated and cultured using the whole bone marrow adherence method. Coffee-conditioned medium was prepared at different concentrations，and the optimal concentration was selected based on alkaline phosphatase （ALP） staining. Changes in osteogenesis-related markers were observed via qRT-PCR and Western blot，and alterations in the gene and protein expression levels of β-catenin were detected. Results Rat BMMSCs were successfully isolated and cultured. ALP activity assay identified 0.1 mg/mL as the optimal concentration of coffee-conditioned medium. Further experiments showed that this concentration had no significant effect on the proliferation of rat BMMSCs during the first 3 days but inhibited their proliferation from Day 5 to 9. Coffee-conditioned medium suppressed the expression of osteogenic markers in rat BMMSCs and concurrently downregulated the expression of proteins related to the Wnt/β-catenin signaling pathway. Conclusion Coffee-conditioned medium can inhibit the osteogenic differentiation of rat BMMSCs，and its mechanism may be associated with the suppression of the Wnt/β-catenin signaling pathway.

Key words: coffee, bone marrow mesenchymal stem cells （BMMSCs）, osteogenic differentiation, Wnt/β-catenin signaling pathway

中图分类号:

R780.1
R36

王洁, 李娜, 于金华. 咖啡通过抑制Wnt/β-catenin通路介导大鼠骨髓间充质干细胞成骨分化[J]. 口腔医学, 2026, 46(5): 337-343.

WANG Jie, LI Na, YU Jinhua. Coffee inhibits osteogenic differentiation of rat bone marrow mesenchymal stem cells via suppression of the Wnt/β-catenin pathway[J]. Stomatology, 2026, 46(5): 337-343.

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基因	引物序列（5'—3'）	GenBank编号
Alp	F：5'-GGAACGGATCTCGGGGTACA-3' R：5'-ATGAGTTGGTAAGGCAGGGT-3'	NM_013059.1
Runx2	F：5'-TTAACGTCAGCAGGAGCAG-3' R：5'-CTTCACCCCCAGGACCAAG-3'	NM_053470.1
Osx	F：5'-GGAGGCACAAAGAAGCCATA-3' R：5'-GGGAAAGGGTGGGTAGTCAT-3'	NM_181374.2
β-catenin	F：5'-GCAGCGACTAAGCAGGAAGG-3' R：5'-TCACCAGCACGAAGGACAGT-3'	NC_086026.1
Gapdh	F：5'-CACTGAGCATCTCCCTCACAA-3' R：5'-GTATTCGAGAGAAGGGAGGGCT-3'	NM_017008.4