地塞米松调控牙本质基质蛋白1在成骨细胞中的表达

Study on Dexamethasone regulation of dentin matrix protein 1 expression in osteoblast

目的研究地塞米松诱导成骨细胞(MC3T3-E1)中牙本质基质蛋白1(DMP1)的表达变化情况。方法利用实时荧光定量PCR反应技术检测不同浓度的地塞米松处理MC3T3-E1细胞后不同时间段细胞中DMP1基因表达变化,通过单因素方差分析法在每个浓度和48 h下对实验组和对照组进行表达差异比较。利用蛋白印迹法(Western blot)检测10^(-6)g/L的地塞米松处理MC3T3-E1细胞后不同时间段的细胞中DMP1蛋白表达变化。结果不同浓度地塞米松分别处理MC3T3-E1成骨细胞DMP1基因表达水平在24、48和72 h均较对照组0 h显著降低(P<0.05),48 h组较24和72 h组显著降低(P<0.05);48 h下10^(-6)g/L组较10^(-7)g/L组显著降低(P<0.05),而与10^(-5)组相比较没有差异(P>0.05)。10^(-6)g/L地塞米松处理MC3T3-E1成骨细胞后,不同时间点的DMP1蛋白表达量均较0 h降低。结论地塞米松抑制成骨细胞MC3T3-E1中DMP1的表达,提示地塞米松可能是通过调控DMP1的表达而影响成骨细胞的骨形成。

Objective To analyze the expression changes of dentin matrix protein 1（DMP1） regulated by Dexamethasone（Dex）in osteoblast cells. Methods Using the quantitative real-time PCR reaction technology to analyze the changes of DMP1 gene expression in MC3T3-E1 cell at various times points under Dex treatment with different concentrations. Then the difference between experimental groups and control groups in 48 h and various concentrations was compared by the data from real-time PCR using One-Way ANOVA statistical method. By means of the western-blot technology,the protein expression changes of DMP1 at various hours in MC3T3-E1 cell under 10-6 g/L Dex treatment were analyzed. Results In MC3T3-E1 cells treated with different concentrations Dex,the DMP1 gene expressions were significantly reduced at 24,48 and 72 h compared with 0 h（P〈0.05）,and 48 h was decreased significantly compared with 24 and 72 h（P〈0.05）;in 48 h the 10^-6 g/L group was decreased significantly compared with 10^-7 g/L （P〈0.05）and with no significant difference compared with 10^-5 g/L（P〈0.05）. In MC3T3-E1 cells under 10^-6 g/L Dex treatment,the DMP1 protein expression of various time points were lower than the baseline （0 h）. Conclusions The expression of DMP1 was suppressed by Dexamethasone in osteoblast MC3T3-E1, which suggested that Dex may influence the development of bone cells by regulate the expression of DMP1.