[1]任格,聂利,杨霞,等.Smad信号通路在骨形成蛋白9促进牙囊干细胞成骨向分化中的研究[J].第三军医大学学报,2018,40(12):1066-1072.
 REN Ge,NIE Li,YANG Xia,et al.Smad signaling pathway regulates bone morphogenetic protein 9induced osteogenic differentiation of rat dental follicle stem cells in vitro[J].J Third Mil Med Univ,2018,40(12):1066-1072.
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Smad信号通路在骨形成蛋白9促进牙囊干细胞成骨向分化中的研究(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第12期
页码:
1066-1072
栏目:
基础医学
出版日期:
2018-06-30

文章信息/Info

Title:
Smad signaling pathway regulates bone morphogenetic protein 9induced osteogenic differentiation of rat dental follicle stem cells in vitro
作者:
任格聂利杨霞黄丽娜王明邱叶李洁仪李丛华
重庆医科大学附属口腔医院,口腔疾病与生物医学重庆市重点实验室,重庆市高校市级口腔生物医学工程重点实验室
Author(s):
REN Ge NIE Li YANG Xia HUANG Lina WANG Ming QIU Ye LI Jieyi LI Conghua

Chongqing Key Laboratory of Oral Disease and Biomedical Sciences, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Stomatological Hospital of Chongqing Medical University, Chongqing, 401147,China

关键词:
牙囊干细胞骨形成蛋白9成骨分化Smad信号通路
Keywords:
rat dental follicle stem cells bone morphogenetic protein 9 osteogenesis   Smad signaling pathway
分类号:
R322.71;R329.2;R394.2
文献标志码:
A
摘要:

目的探索骨形成蛋白9(bone morphogenetic protein 9,BMP9)在诱导大鼠牙囊干细胞(rat dental follicle stem cells,rDFCs)成骨向分化过程中的Smad信号通路调控机制。方法重组腺病毒骨形成蛋白9(recombinant adenoviruses expressing BMP9, AdBMP9)转染纯化的第3代rDFCs后,Real time qPCR、碱性磷酸酶(alkaline phosphatase,ALP)染色及活性检测观察BMP9调节rDFCs早期及中晚期成骨能力,茜素红S染色检测钙盐沉积,Western blot 检测Smad1/5/8磷酸化水平。结果BMP9促进rDFCs成骨因子表达:成骨转录因子Runx2、成骨细胞特异性转录因子Osterix、ALP及骨桥蛋白(osteopontin,OPN)在BMP9刺激组较空白组均明显升高(P<0.05),钙盐沉积及钙结节形成也明显多于空白组,且BMP9促进了Smad1/5/8蛋白磷酸化水平升高。结论BMP9通过激活Smad信号通路,提高Smad1/5/8蛋白磷酸化水平从而促进早中晚期成骨因子及碱性磷酸酶表达,促进了rDFCs成骨向分化。

Abstract:

ObjectiveTo investigate the role of Smad signaling pathway in the regulatory mechanism of bone morphogenetic protein 9 (BMP9)induced osteogenic differentiation of rat dental follicle stem cells (rDFCs). MethodsThe third passage of purified rDFCs were transfected with the recombinant adenoviruses expressing BMP9 (AdBMP9), and the early and late osteogenesis of rDFCs was detected using realtime qPCR, alkaline phosphatase (ALP) staining, ALP activity assay and Alizarin red S staining. The phosphorylation of Smad1/5/8 in the transfected cells was determined using Western blotting. ResultsCompared with the control cells, AdBMP9transfected rDFCs showed significantly enhanced expression of the osteogenic factors including Runx2, Osterix, ALP and osteopontin(OPN) (P<0.05) with also increased calcium deposition and formation of calcium nodules in the early, intermediate and late stages of osteogenesis. Adenovirusmediated BMP9 gene transduction significantly increased the phosphorylation levels of Smad1/5/8 proteins in rDFCs. ConclusionBMP9 can promote the osteogenic differentiation of rDFCs by activating Smad signaling pathway and increasing the phosphorylation level of Smad1/5/8 proteins to enhance the expression of ALP and osteogenic factors in the early, intermediate and late stages of differentiation.

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更新日期/Last Update: 2018-07-02