应力刺激通过Wnt/β⁃catenin通路调控牙骨质细胞的矿化  

Mechanical stress regulates the mineralization of cementocytes through Wnt/β⁃catenin signaling pathway

作　　者：王涵 李天成 陈硕 曾心怡 杨宽 邹淑娟[1] 段沛沛[1] WANG Han;LI Tiancheng;CHEN Shuo;ZENG Xinyi;YANG Kuan;ZOU Shujuan;DUAN Peipei(State Key Laboratory of Oral Dis-eases&National Clinical Research Center for Oral Diseases&Department of Orthodontics,West China Hospital of Stomatology,Sichuan University,Chengdu 610041,China)

机构地区：[1]口腔疾病研究国家重点实验室,国家口腔疾病临床医学研究中心,四川大学华西口腔医院正畸科,四川成都610041 [2]上海交通大学医学院口腔医学院,上海市第九人民医院口腔正畸科,上海200011

出　　处：《口腔医学》2023年第7期600-607,共8页Stomatology

基　　金：四川省科技计划(2021YFS0193);国家自然科学基金(81701005)。

摘　　要：目的牙骨质再矿化对牙根吸收后牙骨质修复至关重要,本文探究应力下Wnt/β⁃catenin通路对牙骨质细胞矿化的调控机制。方法对矿化诱导21 d的牙骨质细胞系IDG⁃CM6细胞加载轻力6 h,通过茜素红染色观察IDG⁃CM6细胞矿化程度,通过实时荧光定量聚合酶链反应(RT⁃qPCR)、蛋白质印迹(Western blot)及免疫荧光检测β⁃catenin及Runx2、Osterix的表达水平。采用生信技术筛选差异表达基因后,通过敲除IDG⁃CM6细胞内β⁃catenin检测矿化程度及Runx2、Osterix表达水平变化。结果应力下,IDG⁃CM6细胞矿化增加,Runx2、Osterix表达水平升高;生信分析及RT⁃qPCR、Western blot、免疫荧光染色表明应力促进β⁃catenin的表达,下调β⁃catenin能逆转应力刺激下的矿化效应。结论应力条件下,Wnt/β⁃catenin通路参与了牙骨质矿化的调控,可能是牙根吸收后牙骨质修复的关键通路。Objective The cementum mineralization is important for cementum repair after root resorption.This study aims to investi⁃gate the role of Wnt/β⁃catenin signaling pathway in regulating the mineralization of cementocyes under mechanical stress.Methods The IDG⁃CM6 cells were subjected to osteogenic⁃induced medium for 21 days and then harvested for establishing in vitro mechanical loading model for 6 h.RNA⁃seq technique was used to screen the differentially expressed genes in IDG⁃CM6 cells under stress.Alizarin red staining was applied to observe the mineralization of IDG⁃CM6 cells,and the expression levels ofβ⁃catenin and osteogenic factors,Runx2 and Osterix,were detected by real⁃time fluorescence quantitative polymerase chain reaction(RT⁃qPCR)and Western blot.The expressions ofβ⁃catenin and Runx2 were also observed by immunofluorescence staining.After knockdown ofβ⁃catenin in IDG⁃CM6 cells by siRNA technology,the mineralization of IDG⁃CM6 cells and the expression levels of Runx2 and Osterix were detected under mechanical stress.Results The mineralization of IDG⁃CM6 cells and the expression levels of Runx2 and Osterix were increased under mechanical stress.The bioinformatic analysis,RT⁃qPCR,Western blot and immunofluorescence staining showed that mechanical stress could promoteβ⁃catenin expression and knockdown ofβ⁃catenin could reverse the mineralization promotion effect produced by mechani⁃cal stress.Conclusion Wnt/β⁃catenin pathway is involved in the regulation of cementum mineralization and plays an important role in cementum mineralization after root resorption.

关 键 词：牙骨质细胞 牙骨质稳态 Wnt/β⁃catenin通路 矿化 

分 类 号：R783.5[医药卫生—口腔医学]