摘要
目的研究Osterix(Osx)过表达对人牙周膜细胞受力后骨向分化的影响,探讨Osx与正畸牙周组织骨改建的关系。方法采用组织块法培养人牙周膜细胞,用重组质粒pcDNA3.1 flag-Osx转染人牙周膜细胞。采用逆转录聚合酶链反应(RT-PCR)和Western blot方法检测未转染组、转染空质粒组、转染Osx组细胞Osx mRNA和蛋白表达水平;并观察核心结合因子α1(Cbfα1)、碱性磷酸酶(ALP)、骨桥素(OPN)、骨钙素(OC)、骨涎蛋白(BSP)和a1(Ⅰ)型胶原蛋白(ColⅠ)的mRNA表达情况。3组细胞加载6 h离心力后,观察上述检测指标的变化。结果转染24 h后,相对未转染组,转染空质粒组Osx mRNA和蛋白水平无明显变化(P>0.05);而转染Osx组Osx mRNA和蛋白表达水平明显升高(P<0.01),同时5个成骨标志基因的mRNA表达亦均明显升高(P<0.05,P<0.01)。加力6 h后,3组Osx和成骨标志基因表达水平均明显升高,但是转染Osx组Osx mRNA和蛋白表达增强更加显著,增加量约为其他两组的2倍,其ALP、OPN、OC、BSP和ColⅠ的mRNA表达亦升高更显著。结论 Osx过表达可以促进人牙周膜细胞在机械力作用下向成骨样细胞分化。Osx可能通过调控多种成骨基因的表达,从而在正畸牙周组织的骨改建过程中发挥着重要的作用。
Objective To investigate the effects of Osterix（Osx） overexpression on the osteogenic differentiation of human periodontal ligament cells in response to mechanical force. Methods Human periodontal ligament cells were isolated and cultured in vitro with explant method. Cells were transfected with either an Osx expression vector pcDNA3.1 flag-Osx or the mock control vector pcDNA3.1 flag. Then, cells were centrifuged for 6 h. After transfection and cen- trification, the expression of Osx mRNA and protein in untransfected cells, mock-transfected cells and Osx-transfected cells were measured by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot respectively. Furthermore, the changes of mRNA expressions of core-binding factor cd （Cbfctl）, alkaline phosphatase （ALP）, osteopontin （OPN）, osteocalcin（OC）, bone sialoprotein（BSP） and collagen protein al（Col I ） genes were measured to assess the differentiation of human periodontal ligament cells. Results At 24 h after transfection, Osx mRNA and protein level increased significantly in Osx-transfeeted cells（P〈O.O1）, while there were no significant difference in Osx mRNA and protein levels between mock-transfected cells and untransfected cells（P〉0.05）. Simultaneously, the upregulated mRNA expressions of all the five osteogenic genes were observed （P〈0.05, P〈0.01）. After 6 h of mechanical stimulation, a sig- nificant increase in Osx expression was shown in all three groups. However, compared to mock-transfected and untransfected cells, Osx-transfected cells further showed the highest Osx mRNA and protein expression level. Furthermore, the mRNA expressions of all five osteogenic markers in Osx-transfected cells also exhibited the greater increase and showed the highest levels. Conclusion The overexpression of Osx promotes the mechanical stress-induced osteogenicdifferentiation of human periodontal ligament cells. Osx may be essential for mechanical stress-induced differentiation of human periodontal ligam
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2013年第2期199-204,共6页
West China Journal of Stomatology
基金
国家自然科学基金资助项目(31000432)
天津医科大学科学基金资助项目(2008ky09)
关键词
OSTERIX
过表达
人牙周膜细胞
机械刺激
成骨分化
Osterix
overexpression
human periodontal ligament cells： mechanical stimulation
osteogenicdifferentiation
