Wei Xi, Wu Li-ping, Ling Jun-qi, Liu Lu
Department of Operative Dentistry and Endodontics, Guanghua School and Hospital of Stomatology & Institute of Stomatological Research, Sun Yat-sen University, Guangzhou 510055, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2009 Sep;44(9):524-8.
To investigate the expression of matrix extracellular phosphorylated protein (MEPE) in human dental pulp cells (hDPC) undergoing odontogenic induction and explore the role of MEPE in odontoblast-like differentiation.
hDPC were isolated by enzymatic digestion and preceded to odontogenic induction for 7, 14 and 21 days respectively. hDPC before induction served as controls. The expressions of MEPE, dentin sialophosphoprotein (DSPP)/dentin sialoprotein (DSP), bone sialoprotein (BSP) and collagen type I were determined by quantitative real-time RT-PCR and Western blotting.
The mRNA levels of MEPE, DSPP, BSP and type I collagen were increased in a time-dependent manner as hDPC were induced along odontoblastic lineage. Statistical differences were detected for MEPE and BSP mRNA expressions in induced hDPC compared with control group (P < 0.001). For DSPP and type I collagen, the mRNA levels in the induced groups were (12 943.33 + or - 3805.73) and (250.55 + or - 31.86) respectively, which were significantly higher than those in control group on days 21 (P < 0.05). Western blotting also revealed the increased expressions of MEPE, DSP, BSP and type I collagen in the induced DPC.
hDPC showed analogously temporal expressions of MEPE, DSPP/DSP, BSP and collagen type I while differentiating along odontoblast lineage. MEPE may play an important role in the odontogenic differentiation of hDPC and may be a potential marker of odontoblast-like differentiation.
研究基质细胞外磷酸化蛋白(MEPE)在人牙髓细胞(hDPC)成牙诱导过程中的表达情况,并探讨MEPE在成牙本质细胞样分化中的作用。
采用酶消化法分离hDPC,分别进行7天、14天和21天的成牙诱导。诱导前的hDPC作为对照。通过定量实时逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测MEPE、牙本质涎磷蛋白(DSPP)/牙本质涎蛋白(DSP)、骨涎蛋白(BSP)和I型胶原的表达。
随着hDPC沿成牙本质细胞谱系诱导,MEPE、DSPP、BSP和I型胶原的mRNA水平呈时间依赖性增加。与对照组相比,诱导后的hDPC中MEPE和BSP的mRNA表达有统计学差异(P<0.001)。对于DSPP和I型胶原,诱导组的mRNA水平分别为(12 943.33±3805.73)和(250.55±31.86),在第21天时显著高于对照组(P<0.05)。蛋白质免疫印迹法也显示诱导后的DPC中MEPE、DSP、BSP和I型胶原的表达增加。
hDPC在沿成牙本质细胞谱系分化时,MEPE、DSPP/DSP、BSP和I型胶原呈现类似的时间表达模式。MEPE可能在hDPC的成牙分化中起重要作用,可能是成牙本质细胞样分化的潜在标志物。