骨形态发生蛋白2/Osterix信号通路调控的前成骨细胞分化  

Bone morphogenetic proteins-2/Osterix signaling pathway regulates the differentiation of preosteoblasts

作　　者：李成琳[1] 陈书兰[2] 任伟伟[3] 

机构地区：[1]潍坊医学院口腔医学院,山东省潍坊市261000 [2]青岛市市立医院口腔医学中心,山东省青岛市266071 [3]湖北医药学院附属东风口腔医院儿童口腔科,湖北省十堰市442000

出　　处：《中国组织工程研究》2016年第24期3581-3587,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:研究发现Osterix基因的表达能够受到骨形态发生蛋白2信号通路的调控,从而调节骨的形成发育。目的:分析骨形态发生蛋白2/Osterix信号通路对小鼠前成骨细胞分化的调控作用。方法:将骨形态发生蛋白2作用于前成骨细胞,利用实时定量RT-PCR法及免疫印迹法检测不同时间段Osterix m RNA和蛋白的表达水平;构建pc DNA3.1/myc-Osterix真核表达载体,并转染至前成骨细胞,利用实时定量RT-PCR法检测转染Osterix真核表达载体和骨形态发生蛋白2作用后碱性磷酸酶、骨涎蛋白、细胞外基质磷酸化糖蛋白(MEPE)的m RNA表达水平。结果与结论:(1)实时定量RT-PCR法检测结果表明骨形态发生蛋白2上调Osterix m RNA在小鼠前成骨细胞中的表达水平,并在24 h时上调作用最为显著;(2)免疫印迹法检测骨形态发生蛋白2显著上调Osterix蛋白的表达水平;(3)成功构建了pc DNA3.1/myc-Osterix真核表达载体,在转染小鼠前成骨细胞和以骨形态发生蛋白2作用后检测到碱性磷酸酶、骨涎蛋白、细胞外基质磷酸化糖蛋白的m RNA表达显著增强;(4)结果说明,骨形态发生蛋白2通过上调小鼠前成骨细胞中Osterix的表达来调控碱性磷酸酶、细胞外基质磷酸化糖蛋白等成骨标志基因的合成,表明骨形态发生蛋白2/Osterix这一信号通路在骨发育过程中具有重要作用。BACKGROUND：Bone formation and development are reported to be regulated by bone morphogenetic protein 2（BMP2）-induced Osterix expression.OBJECTIVE：To investigate the regulatory effect of BMP2/Osterix signaling pathway on differentiation of preosteoblasts in mice.METHODS：m RNA and protein expression of Osterix was determined by real-time RT-PCR and western blot assay,respectively at various time points after mouse preosteoblasts were treated with BMP2.pc DNA3.1/myc-Osterix eukaryotic expression vector was constructed and transducted into preosteoblasts,and then m RNA expression of alkaline phosphatase,bone sialoprotein,and matrix extracellular phosphoglycoprotein was detected by real-time RT-PCR after transduction and BMP2 treatment.RESULTS AND CONCLUSION：Osterix m RNA expression was up-regulated when treated with BMP2 in mouse preosteoblasts,and reached the peak at 24 hours.In addition,the protein expression of Osterix was increased after BMP2 treatment.Alkaline phosphatase,bone sialoprotein,and matrix extracellular phosphoglycoprotein mR NA expression was up-regulated after transfection of mouse preosteoblasts with pc DNA3.1/myc-Osterix eukaryotic expression vector and BMP2 treatment.Our results indicate that BMP2 regulates the synthesis of genetic markers of osteogenesis,such as alkaline phosphatase,matrix extracellular phosphoglycoprotein via up-regulating Osterix expression in mouse preosteoblasts,suggesting BMP2/Osterix signaling pathway plays a critical role in bone development.

关 键 词：骨形态发生蛋白质类 信号传导 成骨细胞 组织工程 组织构建 骨细胞 骨形态发生蛋白2 OSTERIX 前成骨细胞 实时定量RT-PCR 信号通路 免疫印迹 

分 类 号：R318[医药卫生—生物医学工程]