经典Wnt信号通路在牙周膜细胞成骨分化过程中的调控

发布时间：2019-04-19 11:20

【摘要】：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14d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24h、48h、72h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(P0.01);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(P0.01)。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。
[Abstract]:Aim: to investigate the role of classical Wnt/ 尾-catenin signaling pathway in osteoblast differentiation of periodontal ligament mixed cell group. Methods: the mixed periodontal ligament cells were cultured by tissue block-binding enzyme digestion in vitro. The expression of classical Wnt signaling pathway in periodontal ligament cells was confirmed by RT-PCR,real-time PCR. The expression position and expression of 尾-catenin were detected by cellular immunofluorescence analysis. The classical Wnt signaling pathway in periodontal ligament cells was activated by different concentrations of Licl, and the corresponding induction culture was carried out. After culture for 14 days, alizarin red staining was used to observe the formation of mineralized nodules and alkaline phosphatase activity to detect the activity of ALP, and the effects of classical Wnt signaling pathway on the proliferation of periodontal ligament cells were detected by CCK-8 for 24 h, 48 h and 72 h after stimulation with Licl. Single factor repeated measurement of variance analysis was used to compare the differences. Results: the classical Wnt/ 尾-catenin signaling pathway was obviously expressed in periodontal ligament mixed cell group, which could be activated by Licl. The accumulation of 尾-catenin in the cells resulted in downstream changes. Compared with the control group, the activation of classical Wnt/ 尾-catenin signaling pathway resulted in a decrease in mineralized nodules and a significant decrease in ALP activity during the mineralization of periodontal ligament cells (P0.01). Classical Wnt pathway significantly inhibited the proliferation of mixed periodontal ligament cells (P0.01). Conclusion: the classical Wnt/ 尾-catenin signaling pathway inhibits the mineralized osteogenesis of the mixed periodontal ligament cell group and inhibits the proliferation of the cell group.
【作者单位】：武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培训基地和口腔生物医学教育部重点实验室;武汉大学口腔医院牙周科;
【基金】：国家自然科学基金项目(81170933;81570946) 湖北省自然科学基金(2015CFB259) 武汉市科技局青年晨光计划(2014072704011255)
【分类号】：R781.4

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