目的 观察Pax9对小鼠帽状期和钟状期牙胚牙乳头细胞的作用.方法 培养胚胎14.5d和16.5d小鼠下颌第一磨牙牙胚的牙乳头细胞.转染 Pax9 siRNA至牙乳头细胞中敲低 Pax9的表达,CCK8检测转染后24、48、72和96h细胞的增殖情况,Real-time PCR检测转染后Msx1、Bmp2、Bmp4的表达变化.在转染Pax9 siRNA的同时进行矿化诱导培养7d,然后检测Alp、Dmp1和Dspp的表达情况.结果 敲低Pax9表达后,牙乳头细胞的增殖能力减弱;Msx1的表达水平下降,而Bmp2和Bmp4的表达水平升高;牙本质形成相关基因-Alp、Dmp1和Dspp的表达水平升高,牙乳头细胞的矿化能力增强.结论 Pax9参与调控小鼠帽状期和钟状期牙乳头细胞的增殖和成牙本质分化,同时调控下游基因Msx1、Bmp2和Bmp4的表达.%Objective To investigate the roles of Pax9 on regulating mouse dental pallia cells derived from cap and bell stage. Methods Dental pallia cells of mouse mandibular first molar were isolated and cultured from embryonic day(E) 14.5 and 16.5 fetal mice in vitro. Pax9 siRNA was transfected into dental pallia cells in order to knockdown Pax9 expression. After transfection,cell proliferation was determined by CCK8 assay at 24,48,72 and 96 hours. Twenty-four hours after transfection, real-time PCR was employed to detect Msx1, Bmp2 and Bmp4 expression. The differentiation medium was employed to induce the odontoblast differentiation of dental pallia cells. Seven days after differentiation induction and transfection, the expression levels of odontoblast differentiation related gene, Alp, Dmp1, and Dspp were examined. Results After Pax9 down-regulation, the proliferation capacity of dental pallia cells was significantly reduced, and Msx1 expression was decreased,while Bmp2 and Bmp4 expression was increased. The expression levels of odontoblast differentiation related genes,Alp,Dspp and Dmp1 were up-regulated. The odontoblast differentiation potential was elevated after Pax9 down-regulation. Conclusion Pax9 regulates the proliferation and odontoblast differentiation of dental pallia cells derived from cap and bell stage.
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