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p75NTR在大鼠外胚间充质干细胞不同融合状态中的表达差异与矿化相关性研究
Expression difference and mineralization correlation of p75NTR in rat ectomesenchymal stem cells at different fusion states
- 第三军医大学学报 2018年40卷第9期 页码：753-760
- 作者机构：
1.400042 重庆, 陆军军医大学(第三军医大学)第三附属医院(野战外科研究所)口腔科
2.710032 西安, 空军军医大学(第四军医大学)口腔医院牙周病黏膜病科
- 作者简介：
[ 温秀杰, E-mail:wenxiujie@tom.com WEN Xiujie, E-mail:wenxiujie@tom.com
- 基金信息：国家自然科学基金面上项目(8140032);军事口腔医学国家重点实验室开放课题(2014KB14)
DOI：10.16016/j.1000-5404.201711164
中图分类号： R321.5;R329.2;R394.2收稿：2017-11-19，
修回：2018-02-22，
纸质出版：2018-05-15
- 稿件说明：
移动端阅览
肖勇, 李刚, 王莹莹, 等. p75NTR在大鼠外胚间充质干细胞不同融合状态中的表达差异与矿化相关性研究[J]. 第三军医大学学报, 2018,40(9):753-760.
Yong XIAO, Gang LI, Yingying WANG, et al. Expression difference and mineralization correlation of p75NTR in rat ectomesenchymal stem cells at different fusion states[J]. Journal of Third Military Medical University, 2018, 40(9): 753-760.
肖勇, 李刚, 王莹莹, 等. p75NTR在大鼠外胚间充质干细胞不同融合状态中的表达差异与矿化相关性研究[J]. 第三军医大学学报, 2018,40(9):753-760. DOI： 10.16016/j.1000-5404.201711164.
Yong XIAO, Gang LI, Yingying WANG, et al. Expression difference and mineralization correlation of p75NTR in rat ectomesenchymal stem cells at different fusion states[J]. Journal of Third Military Medical University, 2018, 40(9): 753-760. DOI： 10.16016/j.1000-5404.201711164.
摘要
目的
2
探讨不同融合状态对p75 neurotrophin receptor(p75 NTR)在外胚间充质干细胞(ectomesenchymal stem cells
EMSCs)中的表达影响。
方法
2
用酶消化法获取SD大鼠胚胎12.5 d(E12.5d)EMSCs
以不同细胞密度铺板
贴壁后即获得4种融合状态:半融合状态(1.0×10
4
/cm
2
)、融合前状态(1.5×10
4
/cm
2
)、融合状态(2.5×10
4
/cm
2
)、融合后状态(3.5×10
4
/cm
2
)。检测4种融合状态细胞矿化诱导前后p75NTR、Runt相关转录因子2(Runx2)和碱性磷酸酶(ALP)、Ⅰ型胶原蛋白(Col1)在mRNA水平和蛋白水平的表达
茜素红染色检测钙盐沉积情况。
结果
2
矿化诱导0 d时
p75NTR mRNA和蛋白在融合状态组表达最高
Runx2、ALP、Col1在4种融合状态表达水平无明显差异。矿化诱导3、7 d时
融合状态组p75NTR、Runx2、ALP mRNA表达水平均显著高于其他3组(
P
<
0.05);同时融合状态组p75NTR、Runx2、Col1蛋白表达水平最高
差异具有统计学意义(
P
<
0.05)。矿化诱导21 d时
融合状态组钙盐沉积量高于其他3组(
P
<
0.01)。
结论
2
不同融合状态p75NTR表达差异显著
在融合状态组p75NTR表达最高
此时细胞矿化能力最强
表明p75NTR的表达与矿化呈正相关。
Abstract
Objective
2
To determine the effects of different fusion states on the expression of p75NTR in ectomesenchymal stem cells (EMSCs).
Methods
2
EMSCs were obtained from SD rat fetuses at embryonic day 12.5 (E12.5d) by enzymatic digestion.After being paved at different cell densities and then adherent to the wall
the cells were assigned into 4 fusion states
that is
semi-fusion (1.0×10
4
/cm
2
)
pre-fusion (1.5×10
4
/cm
2
)
fusion state (2.5×10
4
/cm
2
)
and post-fusion state (3.5×10
4
/c
m
2
).The expression of p75NTR
Runt related transcription factor 2 (Runx2)
alkaline phosphatase (ALP) and type Ⅰ collagen (Col1) at mRNA and protein levels were detected in the 4 fusion states before and after mineralized inducement.Alizarin red staining was used to observe calcium salt deposition.
Results
2
Without mineralization inducement
the mRNA and protein expression of p75NTR reached the highest levels in the fusion state group
while no differences were seen in the levels of Runx2
ALP and Col1 in 4 fusion state groups.After mineralization inducement for 3 and 7 d
the mRNA and protein levels of p75NTR
Runx2 and ALP were highest in the fusion state group when compared with the other 3 groups of EMSCs (
P
<
0.05).Mineralization inducement for 21 d resulted in the strongest calcium salt deposition in the fusion cells than the 3 groups at other states (
P
<
0.01).
Conclusion
2
p75NTR expression is significantly different in the EMSCs at different fusion states
and it is the highest in the fusion state.The cells show strongest mineralization capacity at this time
indicating that the expression of p75NTR is positively correlated with the mineralization.
关键词
Keywords
references
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