牙发生相关磷蛋白促进成骨细胞粘附及矿化研究  

Study on Odaph Promoting Osteoblasts Adhesion and Mineralization

作　　者：李明玥 肖爽 栾霖群 张杰 高玉光 张莉 LI Mingyue;XIAO Shuang;LUAN Linqun;ZHANG Jie;GAO Yuguang;ZHANG Li(Institute of Stomatology,Binzhou Medical University,Yantai 264003,China)

机构地区：[1]滨州医学院口腔医学院,山东烟台264003

出　　处：《口腔医学研究》2023年第9期810-814,共5页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:81870738);滨州医学院“临床+X”科技创新项目(编号:BY2021LCX08)。

摘　　要：目的:本研究旨在探讨牙发生相关磷蛋白(odontogenesis-associated phosphoprotein,Odaph)对成骨细胞粘附及矿化的影响。方法:实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测小鼠骨组织中Odaph的表达;Odaph过表达慢病毒感染小鼠胚胎成骨细胞前体细胞MC3T3-E1,qRT-PCR检测Odaph过表达对MC3T3-E1的影响;qRT-PCR、Western blot和免疫细胞化学法分别检测Odaph过表达对粘附标志物Lamininγ2和Integrinβ6表达的影响;Odaph和Lamininγ2质粒共转染至人胚肾细胞-293T(human embryonic kidney cells-293T,HEK-293T),免疫共沉淀验证两者是否存在相互作用;MC3T3-E1经诱导矿化后,qRT-PCR检测矿化相关标志物ALP、OCN和Runx2的表达,碱性磷酸酶(alkaline phosphatase,ALP)染色观察ALP活性。结果:研究显示Odaph在小鼠骨组织和MC3T3-E1中表达;Odaph过表达促进了Lamininγ2和Integrinβ6的表达,且Odaph与Lamininγ2存在相互作用;矿化诱导后Odaph过表达促进ALP、OCN和Runx2的表达,并且增强了ALP活性。结论:Odaph具有促进成骨细胞粘附及矿化的作用。Objective:To explore the influence of odontogenesis-associated phosphoprotein(Odaph)on osteoblast adhesion and mineralization.Methods:The expression of Odaph in mouse bone tissue was detected by quantitative real-time PCR(qRT-PCR).The effect of Odaph overexpression on MC3T3-E1 was detected by qRT-PCR.qRT-PCR,Western blot,and immunocytochemistry were used to detect the effect of Odaph overexpression on the expression of Lamininγ2 and Integrinβ6.Odaph and Lamininγ2 plasmid were cotransfected into human embryonic kidney cells-293T(HEK-293T),and co-immunoprecipitation was used to verify the interaction between Odaph and Lamininγ2.After mineralization induction of MC3T3-E1,the expressions of mineralization-related markers ALP,OCN,and Runx2 were detected by qRT-PCR,and ALP activity was observed by alkaline phosphatase(ALP)staining.Results:Odaph was expressed in mouse bone tissue and MC3T3-E1.Odaph overexpression increased the expression of Lamininγ2 and Integrinβ6,and Odaph interacted with Lamininγ2.After mineralization induction,Odaph overexpression promoted the expression of ALP,OCN,and Runx2,and enhanced ALP activity.Conclusion:Odaph promoted osteoblast adhesion and mineralization.

关 键 词：牙发生相关磷蛋白 MC3T3-E1 粘附 矿化 碱性磷酸酶 

分 类 号：R781[医药卫生—口腔医学]