Prrx2对小鼠磨牙胚发育的影响研究  

The effect of Prrx2 on the development of mouse molar germ

作　　者：朱辙文 韩雪 刘力维 王佐林[1] ZHU Zhewen;HAN Xue;LIU Liwei;WANG Zuolin(Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Department of Oral Implantology,School and Hospital of Stomatology,Tong ji University,Shanghai 200072,China)

机构地区：[1]上海牙组织修复与再生工程技术研究中心,同济大学口腔医学院,同济大学附属口腔医院口腔种植科,上海200072

出　　处：《口腔颌面外科杂志》2023年第3期131-137,共7页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(82000995);国家重点研发计划(2018YFE0202200);中央高校基本业务费专项资金(22120180196)。

摘　　要：目的:探究配对相关同源框2(paired related homeobox 2,Prrx2)在小鼠磨牙胚不同发育时期的表达及其对磨牙胚发育的影响。方法:利用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)分析配对相关同源框1(paired related homeobox 1,Prrx1)和Prrx2在小鼠胚胎12.5 d(E12.5)至E18.5,出生后1、3、7 d(P1、P3、P7)磨牙牙胚中的表达,以及其在E13.5磨牙、毛发、脑、心脏、结肠、肾、肺、唾液腺等组织中的相对表达情况;使用小干扰RNA(small interfering RNA,siRNA)沉默Prrx2基因的表达,通过对牙胚行体外器官培养,观察牙胚发育情况;并利用CCK8实验、细胞划痕实验观察牙胚间充质细胞的增殖和迁移能力。结果:RT-qPCR结果显示,相比其他组织,Prrx2在E13.5小鼠胚胎的磨牙、毛发、唾液腺中的表达量更高;相比于Prrx1,Prrx2磨牙胚发育后期(E16.5~E18.5)的表达水平仍高于初始阶段(E13.5);牙胚体外培养结果显示,沉默Prrx2后,牙胚宽度、高度及牙尖高度分别减少了(21.11±3.49)%、(12.08±5.30)%和(22.34±7.56)%(均P<0.05);CCK8实验、细胞划痕实验结果显示,沉默Prrx2后,牙胚间充质细胞增殖能力无变化(P>0.05)、迁移能力降低(P<0.05)。结论:Prrx2在牙胚形态成形的过程中,承担着重要的作用;Prrx2通过促进磨牙胚间充质细胞的迁移能力,进而影响磨牙胚生长和牙尖发育的进程。Objective:To investigate the expression level of paired related homeobox 2(Prrx2)at different stages of mouse molar germ and its effect on molar germ development.Methods:Real-time quantitative polymerase chain reaction(RT-qPCR)was used to analyze the relative expression of paired related homeobox 1(Prrx1)and Prrx2 in E12.5 to E18.5,P1,P3,P7,and E13.5 molar germ,hair,brain,heart,colon,kidney,lung,salivary gland and other tissues;small interfering RNA(siRNA)was used to silence the expression of Prrx2,then tooth germ development was observed via vitro culture systems;CCK8 test and cell scratch test were utilized to observe the proliferation and migration ability of molar germ mesenchymal cells.Results:According to the results of RT-qPCR,comparing with Prrx2 in other organs,it was highly expressed in the molars,hair and salivary glands of E13.5 mice;comparing with Prrx1,the expression level of Prrx2 at the late stage of tooth germ development(E16.5-E18.5)is still higher than that at the initial stage of development(E13.5);the results of in vitro tooth germ culture demonstrated that after silencing Prrx2,tooth germ width,height and tooth cusp height were reduced by(21.11±3.49)%,(12.08±5.30)%and(22.34±7.56)%(all P<0.05),respectively;CCK8 test and cell scratch test showed that the proliferation rate of tooth germ mesenchymal cells did not change after silencing Prrx2(P>0.05),but the migration ability was suppressed(P<0.05).Conclusion:Prrx2 plays an important role in the process of molar germ morphogenesis;Prrx2 affects the growth of molar germ and the process of tooth cusp development via promoting the migration ability of molar germ mesenchymal cells.

关 键 词：牙胚发育 Prrx2 体外培养 小鼠 

分 类 号：R782[医药卫生—口腔医学]