经典Wnt信号通路在牙周膜细胞成骨分化过程中的调控

目的: 探讨经典Wnt/β-catenin信号通路对牙周膜混合细胞群成骨分化过程中的调控作用。方法: 体外组织块结合酶消化法培养牙周膜混合细胞群,通过RT-PCR、real-time PCR证实经典Wnt信号通路在牙周膜细胞中的表达,并运用细胞免疫荧光分析检测了β-catenin的表达位置及表达量;通过不同浓度的Licl激活牙周膜细胞中经典Wnt信号通路,并进行相应的诱导培养。培养14 d后,茜素红染色观察矿化结节形成情况,碱性磷酸酶活性检测ALP活性;通过CCK-8检测Licl刺激24 h、48 h、72 h后经典Wnt信号通路对牙周膜细胞增殖的影响。以单因素重复测量方差分析比较差异。结果: 经典Wnt/β-catenin信号通路在牙周膜混合细胞群中明显表达,可以通过Licl激活该信号通路,使β-catenin在细胞中累积引起下游变化;与对照组相比较,经典Wnt/β-catenin信号通路的激活引起牙周膜细胞矿化过程中矿化结节的减少,显著降低了ALP活性(P<0.01);经典Wnt通路对牙周膜混合细胞群增殖表现为显著的抑制作用(P<0.01)。结论: 经典Wnt/β-catenin信号通路抑制了牙周膜混合细胞群的矿化成骨过程,并抑制该细胞群的增殖。

Effect of Canonical Wnt Signalling Pathway on the Osteogenic Differentiation and Proliferation of PDLCs.

Objective: To investigate the effects of canonical Wnt signalling pathway on the osteogenic osteogenic differentiation of human periodontal ligament cells (HPDLCs). Methods: HPDLCs were obtained through in vitro culture combined with enzyme digestion method and tissue pieces culture method. The existence of canonical Wnt signalling pathway was examined by RT-PCR and immunocytochemistry. Lithium chloride (LiCl) was applied during osteogenesis, and both Alizarin red staining and Alkaline phosphatase activity detection were used to investigate influence of Wnt/β-catenin signaling pathway on the osteogenic differentiation of PDLCs．CCK-8 assay was performed to study the effect of activated canonical Wnt signalling pathway on the proliferation of PDLCs. Results: The expression of canonical Wnt signaling pathway conponents were strongly expressed in PDLCs. Negative effects of canonical Wnt signaling pathway on osteoblast differentiation were proved by Alizarin red staining and Alkaline phosphatase activity detection (P<0.01). CCK-8 assay also showed the inhibition of canonical Wnt signaling pathway on the proliferation of PDLCs. Conclusion: Activation of canonical Wnt pathway may inhibit the osteogenic differentiation and proliferation of PDLCs.