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Deltex-1对骨髓间充质干细胞增殖及其向平滑肌细胞分化的影响
Deltex-1 overexpression suppresses proliferation and promotes differentiation of bone marrow mesenchymal stem cells into smooth muscle cells
in vitro - 第三军医大学学报 2018年40卷第2期 页码：109-115
- 作者机构：
1.400021 重庆，重庆市妇幼保健院
2.400038 重庆，陆军军医大学(第三军医大学)第一附属医院妇产科
- 作者简介：
[徐惠成，E-mail：xhuicheng@yahoo.com.cn XU Huicheng, E-mail: xhuicheng@yahoo.com.cn
- 基金信息：国家自然科学基金面上项目(30772309)
DOI：10.16016/j.1000-5404.201708201
中图分类号： R322.74;R329.24;R394.2收稿：2017-08-30，
修回：2017-10-17，
纸质出版：2018-01-30
- 稿件说明：
移动端阅览
王荥, 王延洲, 刘禄斌, 等. Deltex-1对骨髓间充质干细胞增殖及其向平滑肌细胞分化的影响[J]. 第三军医大学学报, 2018,40(2):109-115.
Ying WANG, Yanzhou WANG, Lubin LIU, et al. Deltex-1 overexpression suppresses proliferation and promotes differentiation of bone marrow mesenchymal stem cells into smooth muscle cells
in vitro [J]. Journal of Third Military Medical University, 2018, 40(2): 109-115.王荥, 王延洲, 刘禄斌, 等. Deltex-1对骨髓间充质干细胞增殖及其向平滑肌细胞分化的影响[J]. 第三军医大学学报, 2018,40(2):109-115. DOI： 10.16016/j.1000-5404.201708201.
Ying WANG, Yanzhou WANG, Lubin LIU, et al. Deltex-1 overexpression suppresses proliferation and promotes differentiation of bone marrow mesenchymal stem cells into smooth muscle cells
in vitro [J]. Journal of Third Military Medical University, 2018, 40(2): 109-115. DOI： 10.16016/j.1000-5404.201708201.
摘要
目的
2
探讨Deltex-1对骨髓间充质干细胞(bone marrow mesenchymal stem cells
bMSCs)增殖及其向平滑肌细胞(smooth muscle cells，SMCs)分化的影响。
方法
2
分离培养大鼠骨髓bMSCs，采用流式细胞仪检测鉴定。通过Deltex-1过表达腺病毒载体pAd/Deltex-1感染bMSCs后，采用CCK-8(cell count kit-8)法检测Deltex-1过表达对bMSCs增殖的影响; 将未感染、经Deltex-1病毒感染及经空病毒感染的bMSCs与SMCs共培养，采用免疫荧光细胞化学、RT-PCR及Western blot分别检测其bMSCs中SMCs标志物平滑肌肌球蛋白重链(smooth muscle myosin heavy chain，SM-MHC)及Notch信号通路相关因子Notch-1的表达，单纯培养的bMSCs、SMCs的相同检测作正常对照。
结果
2
成功分离、培养大鼠bMSCs。CCK-8检测证实：与对照比较，经Deltex-1病毒感染的bMSCs生长减慢，48 h开始显得更为明显(
P
<
0.05，
P
<
0.01);免疫荧光细胞化学、RT-PCR及Western blot检测显示：与对照比较，经Deltex-1病毒感染的bMSCs与SMCs共培养后显著表达SMCs的标志物SM-MHC，较弱表达Notch信号通路相关因子Notch-1(
P
<
0.01)。
结论
2
Deltex-1过表达可抑制bMSCs的增殖，并促进其向SMCs分化。
Abstract
Objective
2
To investigate the effect of Deltex-1 overexpression on the proliferation of bone marrow mesenchymal stem cells (bMSCs) and their differentiation into smooth muscle cells (SMCs) in vitro.
Methods
2
Rat bMSCs were isolated
cultured and identified. The effect of Deltex-1 overexpression on the proliferation of bMSCs infected with the adenovirus vector pAd/Deltex-1 was observed using cell count kit-8 (CCK-8). In a co-culture system with SMCs
bMSCs infected with pAd/Deltex-1 or an empty viral vector or with no treatment were examined for the expression of smooth muscle myosin heavy chain (SM-MHC
a marker of SMCs) using immunofluorescence cytochemistry staining
RT-PCR and Western blotting
with bMSCs and SMCs without treatment in separate cultures as normal controls.
Results
2
Compared with the control cells
bMSCs infected by the adenovirus vector pAd/Deltex-1 exhibited significantly lowered growth rate
which was especially obvious at 48 h after the infection (
P
<
0.05
0.01). The
Results
2
of immunofluorescence cytochemistry
real-time PCR and Western blotting all showed that bMSCs infected with the Deltex-1 vector and co-cultured with SMCs expressed a significantly increased level of SM-MHC with a lowed expression of Notch-1 (
P
<
0.01).
Conclusion
2
Deltex-1 overexpression causes suppression of bMSC proliferation and promotes the differentiation of bMSCs into smooth muscle cells in vitro.
关键词
Keywords
references
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