Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells <italic style="font-style: italic">in vitro</italic>

Yingchun XIE; Xiaolin TU

doi:10.16016/j.1000-5404.202001048

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Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells in vitro

Basic Medicine | 更新时间：2026-01-09

- Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells in vitro
- Journal of Third Military Medical University Vol. 42, Issue 9, Pages: 891-898(2020)
- 作者机构：
  
  400016 重庆，重庆医科大学生命科学研究院骨发育与再生平台
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81672118)
- DOI：10.16016/j.1000-5404.202001048
  CLC： R322.71;R329.2;R394-33
- Received：07 January 2020，
  
  Revised：2020-02-23，
  
  Published：15 May 2020
- 稿件说明：
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Yingchun XIE, Xiaolin TU. Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(9): 891-898.
DOI：

Yingchun XIE, Xiaolin TU. Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells in vitro[J]. Journal of Third Military Medical University, 2020, 42(9): 891-898. DOI： 10.16016/j.1000-5404.202001048.

摘要

目的

探究体外激活骨细胞Notch信号对骨髓基质细胞(bone marrow stromal cells

BMSCs)成骨分化的影响。

方法

设置Cre重组腺

病毒(Ad-Cre)及对照腺病毒GFP(Ad-GFP)，分别转染Rosa

Notch

小鼠骨细胞后与野生C57BL/6小鼠BMSCs共培养为Ad-Cre组及Ad-GFP组，设置仅BMSCs为Blank组。碱性磷酸酶染色及生化定量测定碱性磷酸酶(ALP)；qPCR检测转染后骨细胞Notch信号靶基因及共培养产物成骨相关标志物、Notch信号配体及成血管相关标志物，茜素红染色检测成骨诱导21 d钙盐沉积水平。

结果

Ad-Cre成功激活Rosa

Notch

小鼠骨细胞Notch信号，ALP染色及生化定量结果显示Ad-Cre组ALP表达相较Ad-GFP组及Blank组显著降低(

0.05)，Ad-GFP组与Blank组间无统计学差异；qPCR结果显示Ad-Cre组共培养产物成骨标志物ALP、核心结合因子a1(Runx2)、特异性骨转化因子(Ostrix)及Notch配体Dll4 mRNA转录水平相较Ad-GFP组及Blank组显著降低(

0.05)，Jag1 mRNA转录水平显著降低(

0.05)；血管内皮生长因子(VEGF)、低氧诱导因子(hypoxia-inducible factor 1

，HIF1

)、血小板内皮细胞黏附分子(platelet endothelial cell adhesion molecule-1

CD31/PECAM-1)及内皮粘蛋白(endomucin

EMCN)mRNA转录水平相较Ad-GFP组及Blank组显著增高(

0.05)；茜素红染色结果显示Ad-Cre组在成骨诱导培养基作用21 d后钙盐沉积水平相较Ad-GFP组及Blank组显著降低。

结论

体外激活骨细胞Notch信号抑制骨髓基质细胞成骨分化。

Abstract

Objective

To investigate the effect and underlying mechanism of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells (BMSCs).

Methods

The osteocytes derived from Rosa

Notch

mice were infected with recombinant adenovirus expressing Cre or GFP respectively

and then the obtained cells were co-cultured with BMSCs isolated from wild-type C57BL/6 mice

and named as Ad-Cre group and Ad-GFP group

with a BMSCs group without any treatment as blank control. Alkaline phosphatase (ALP) staining and detection of ALP relative activity were used to measure the expression of ALP. The mRNA expression levels of the target genes in osteocyte of Rosa

Notch

and ligands of Notch signaling

osteogenic markers

and angiogenic makers in the co-cultured product were detected by qPCR. Alizarin red staining was applied to test the matrix mineralization on day 21.

Results

Infection of

recombinant adenovirus Ad-Cre could successfully activate Notch signaling. ALP staining and detection of ALP activity showed that the expression level of ALP was significantly lower in the Ad-Cre group than the Ad-GFP group and Blank group (

0.05)

and there was no statistical difference in the level between the latter 2 groups. The results of qPCR indicated that the mRNA levels of osteogenic makers ALP

Osterix

Runx2 and Notch ligand Dll4 were deceased significantly

while those of Jag1

VEGF

hypoxia-inducible factor 1

(HIF1

)

platelet endothelial cell adhesion molecule-1 (CD31/PECAM-1)

and endomucin (EMCN) were statistically increased in the Ad-Cre group when compared with the Ad-GFP group and Blank group (

0.05). Alizarin red staining displayed that the Ad-Cre group had more calcium deposition in 21 d after co-culture than the Ad-GFP group and blank group (

0.05).

Conclusion

Activating Notch signaling in osteocytes inhibits osteogenic differentiation of BMSCs

in vitro.

关键词

Keywords

references

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⁰

Impact of activating Notch signaling in osteocytes on osteogenic differentiation of bone marrow stromal cells in vitro

DOI：10.16016/j.1000-5404.202001048

摘要

Abstract

关键词

Keywords

references