›› 2013, Vol. 33 ›› Issue (6): 366-370.

• 基础与临床研究 • 上一篇    下一篇

RT-PCR检测釉基质蛋白对大鼠骨髓基质细胞成骨相关基因表达影响的研究

吴飞俊1,赵 芳2,裴路2   

  1. 1. 哈尔滨医科大学附属第二医院牙体牙髓科
    2. 哈尔滨医科大学附属第二医院口腔科
  • 收稿日期:2012-10-11 修回日期:2012-11-25 出版日期:2013-06-28 发布日期:2013-07-12
  • 通讯作者: 吴飞俊 E-mail:wufeijun2010@163.com
  • 基金资助:
    细胞-支架材料复合物修复牙槽骨缺失基础研究

Effect of enamel matrix proteins on expression of bone formation-related genes of rat BMSCs were detected by RT-PCR

  • Received:2012-10-11 Revised:2012-11-25 Online:2013-06-28 Published:2013-07-12
  • Contact: WU feijun E-mail:wufeijun2010@163.com

摘要: 目的 研究釉基质蛋白(enamel matrix proteins,EMPs)对体外培养大鼠骨髓基质细胞(bone marrow stromal cells,BMSCs) 成骨相关基因表达的影响。方法 采用乙酸法从猪恒牙胚中提取釉基质蛋白,取3周龄雄性SD大鼠胫骨和股骨骨髓体外培养,取生长良好的第3代细胞加入不同浓度的EMPs(0、25、50、100、200 mg/L)进行培养,MTT法检测BMSCs的增值,用RT-PCR测定成骨相关基因骨涎蛋白(bone sialoprotein, BSP),骨桥蛋白(osteopontin, OPN)和I型胶原 (collagen typeⅠ, COL-Ⅰ)的表达。结果 实验组的细胞增值大于对照组(p<0.05),并且EMPs的浓度在100 mg/L时增殖作用最强;实验组骨相关基因(BSP、COL-Ⅰ)mRNA的表达高于对照组,并且具有浓度依赖性,但是实验组与对照组OPN mRNA的表达没有区别。结论 EMPs通过调节成骨相关基因的表达而促进大鼠BMSCs向成骨细胞分化。

关键词: 釉基质蛋白, 骨髓基质细胞, 基因表达, 成骨相关基因, BMSCs, gene expression, bone formation-related gene

Abstract: Objective To study the effects of EMPs on expression of bone formation-related gene of rat bone marrow stromal cells culutured in vitvo by RT-PCR. Methods Enamel matrix proteins extracted from porcine permanent tooth buds by acidic extraction. Marrow of tibiae and femurs from 3-week-old male SD rats were obtained and cultured in vitro. Well-grown cells at passage 3 were cultured with different concentrations of EMPs(0, 25,50,100,200 mg/L). MTT assay was performed to evaluate the proliferation of BMSCs. The expression of bone formation-related genes BSP, OPN and COL-Ⅰwere detected by RT-PCR. Results Cell proliferation in experimental group was higher than that in control group (P<0.05), and the proliferation was highest when concentration of EMPs was 100 mg/L. The mRNA expression of bone-related genes (BSP、COL-Ⅰ)in experimental group was higher than that in control group, and was in a concentration dependent manner. But, there was no difference between the control group and the experimental group in mRNA expression of OPN gene. Conclusion EMPs enhance osteoblastic differentiation of rat BMSCs by regulation of bone formation-related gene expression.

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