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

任格; 聂利; 杨霞; 黄丽娜; 王明; 邱叶; 李洁仪; 李丛华

doi:10.16016/j.1000-5404.201711134

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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

基础医学 | 更新时间：2026-01-09

- 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
- Smad signaling pathway regulates bone morphogenetic protein 9-induced osteogenic differentiation of rat dental follicle stem cells in vitro
- 第三军医大学学报 2018年40卷第12期页码：1066-1072
- 作者机构：
  
  401147 重庆，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
- 作者简介：
  
  [ 李丛华，E-mail：liconghua1@163.com LI Conghua, E-mail:liconghua1@163.com
- 基金信息：
  
  重庆市卫计委重点项目(2012-1-056);重庆市科委自然科学基金(CSTC 2016jcyjA0415);重庆市卫计委面上项目(2016MSXM049)
- DOI：10.16016/j.1000-5404.201711134
  中图分类号： R322.71;R329.2;R394.2
- 收稿：2017-11-16，
  
  修回：2018-01-03，
  
  纸质出版：2018-01
- 稿件说明：
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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.

Ge REN, Li NIE, Xia YANG, et al. Smad signaling pathway regulates bone morphogenetic protein 9-induced osteogenic differentiation of rat dental follicle stem cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(12): 1066-1072.
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. DOI： 10.16016/j.1000-5404.201711134.

Ge REN, Li NIE, Xia YANG, et al. Smad signaling pathway regulates bone morphogenetic protein 9-induced osteogenic differentiation of rat dental follicle stem cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(12): 1066-1072. DOI： 10.16016/j.1000-5404.201711134.

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(

＜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

Objective

To 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).

Methods

The 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.

Results

Compared with the control cells

Ad-BMP9-transfected rDFCs showed significantly enhanced expression of the osteogenic factors including Runx2

Osterix

ALP and osteopontin(OPN) (

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 prot

eins in rDFCs.

Conclusion

BMP9 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.

关键词

Keywords

references

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