张应力刺激下Wnt／β-catenin信号通路对成牙骨质细胞Runx2表达调控的体外研究

目的研究张应力刺激下Wnt/β-catenin信号通路对成牙骨质细胞Runx2表达的调控。方法以成牙骨质细胞样细胞OCCM30为研究对象,应用FX4000T应力加载装置对其加载张应力,时间为0、3、6、12 h,然后分别使用不同质量浓度的Dikkopf-1（DKK1）处理48 h,采用Western blot法检测β-catenin蛋白的表达水平,实时荧光定量聚合酶链反应法（RT-PCR）检测Runx2 mRNA的表达水平。选取最佳作用时间点（12 h）和最佳阻断剂质量浓度（150 ng·m L^-1）,将样本分为A、B、C、D共4组,A组不加力不加阻断剂,B组加力加阻断剂,C组加力不加阻断剂...

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Published in华西口腔医学杂志 Vol. 33; no. 1; pp. 35 - 39
Main Author 李书琴 杨珊 任嫒姝 戴红卫
Format Journal Article
LanguageChinese
Published 重庆医科大学附属口腔医院正畸科，口腔疾病与生物医学重庆市重点实验室，重庆 401147 2015
Subjects
Runx2
Wnt／β-catenin
成牙骨质细胞
牙根修复
牙根吸收
牙根吸收
牙根修复
成牙骨质细胞
Wnt/β-catenin
Runx2
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ISSN1000-1182
DOI10.7518/hxkq.2015.01.008

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Summary:目的研究张应力刺激下Wnt/β-catenin信号通路对成牙骨质细胞Runx2表达的调控。方法以成牙骨质细胞样细胞OCCM30为研究对象,应用FX4000T应力加载装置对其加载张应力,时间为0、3、6、12 h,然后分别使用不同质量浓度的Dikkopf-1（DKK1）处理48 h,采用Western blot法检测β-catenin蛋白的表达水平,实时荧光定量聚合酶链反应法（RT-PCR）检测Runx2 mRNA的表达水平。选取最佳作用时间点（12 h）和最佳阻断剂质量浓度（150 ng·m L^-1）,将样本分为A、B、C、D共4组,A组不加力不加阻断剂,B组加力加阻断剂,C组加力不加阻断剂,D组不加力加阻断剂,测量β-catenin蛋白和Runx2 mRNA的表达水平。结果 1）张应力加载3、6、12 h后,Runx2 mRNA表达水平和β-catenin蛋白水平与对照组相比均明显升高,差异有统计学意义（P〈0.05）。2）不同质量浓度DKK1处理后,细胞内Runx2 mRNA的表达量呈下降趋势。3）未加载张应力的情况下,DKK1明显抑制了β-catenin蛋白的表达,Wnt/β-catenin通路被抑制;加载张应力情况下,DKK1处理组β-catenin蛋白表达低于未加DKK1处理组;不加阻断剂的情况下,张应力刺激增强了β-catenin蛋白的表达;阻断剂作用后,应力刺激组β-catenin蛋白表达相对较高。结论机械应力刺激下Wnt/β-catenin信号通路可正向调控成牙骨质细胞Runx2基因的表达。
Bibliography:Objective Periodontal tissue remodeling includes remodeling of alveolar bone, periodontal ligament, and cementum. Cementoblast plays a main role in repairing root resorption. Canonical Wnt/β-catenin signaling can promote the odontogenic differentiation in osteoblast. However, the mechanism on how the orthodontic force influences the function of cementoblast and the relationship between the canonical Wnt/β-catenin signaling and Rtmx2 of cementoblast are＇not yet known. The aim of this study is focus on this relationship. Methods OCCM30 cementoblasts were subjected to mechanical strain by four-point bending system with tension stress for 0, 3, 6, and 12 h. They were pretreated with different concentrations of Dikkopf-1 （DKK1） for 48 h. Western blot analysis was performed to detect the β-catenin levels in the nucleus. Rtmx2 mRNA was observed by real-time quantitative polymerase chain reaction （RT-PCR）. OCCM30 cementoblasts were then pretreated with 150 ng·mL^-1 DKK1 for 48 h and subjected to mechanical strain by
ISSN:1000-1182
DOI:10.7518/hxkq.2015.01.008