牙发生相关磷蛋白促进成骨细胞粘附及矿化研究

doi:10.13701/j.cnki.kqyxyj.2023.09.009

口腔医学研究 ›› 2023, Vol. 39 ›› Issue (9): 810-814.DOI: 10.13701/j.cnki.kqyxyj.2023.09.009

牙发生相关磷蛋白促进成骨细胞粘附及矿化研究

李明玥, 肖爽, 栾霖群, 张杰, 高玉光, 张莉^*

滨州医学院口腔医学院山东烟台 264003

收稿日期:2023-05-05 出版日期:2023-09-28 发布日期:2023-09-25
通讯作者: *张莉,E-mail:zhangli@bzmc.edu.cn
作者简介:李明玥(1997~ ),女,山东枣庄人,硕士在读,主要从事矿化组织发育与再生研究。
基金资助:
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)

Study on Odaph Promoting Osteoblasts Adhesion and Mineralization

LI Mingyue, XIAO Shuang, LUAN Linqun, ZHANG Jie, GAO Yuguang, ZHANG Li^*

Institute of Stomatology, Binzhou Medical University, Yantai 264003, China

Received:2023-05-05 Online:2023-09-28 Published:2023-09-25

摘要/Abstract

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。

Abstract: 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.

Key words: odontogenesis-associated phosphoprotein, MC3T3-E1, adhesion, mineralization, alkaline phosphatase

李明玥, 肖爽, 栾霖群, 张杰, 高玉光, 张莉. 牙发生相关磷蛋白促进成骨细胞粘附及矿化研究[J]. 口腔医学研究, 2023, 39(9): 810-814.

LI Mingyue, XIAO Shuang, LUAN Linqun, ZHANG Jie, GAO Yuguang, ZHANG Li. Study on Odaph Promoting Osteoblasts Adhesion and Mineralization[J]. Journal of Oral Science Research, 2023, 39(9): 810-814.

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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

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