[肿瘤坏死因子-α对人乳牙脱落干细胞促进破骨细胞生成能力的影响]

[Effect of TNF-α on the ability of stem cells from human exfoliated deciduous teeth to promote osteoclastogenesis].

作者信息

Wang Yu-Chen, Wang Chen, Liu Na, Cai Chuan, Liu Wei, Xu Lu-Lu

机构信息

Department of Orthodontics, General Hospital of Chinese PLA. Beijing 100853, China. E-mail:

出版信息

Shanghai Kou Qiang Yi Xue. 2018 Oct;27(5):449-454.

PMID:30680384

Abstract

PURPOSE

To investigate the effect of tumor necrosis factor-α (TNF-α) on the ability of stem cells from human exfoliated deciduous teeth (SHED) to promote osteoclastogenesis.

METHODS

SHED were obtained from deciduous teeth and isolated, purified, cultured in vitro. An indirect co-culture system of SHED and osteoclast precursor peripheral blood mononuclear cells (PBMCs) was established. The expression of osteoclastic gene from PBMCs and NF-κB from SHED were determined after treatment with TNF-α (0, 5, 10, 50, 100 ng/mL) by real-time RT-PCR and Western blot. SPSS 19.0 software package was used for statistical analysis.

RESULTS

Under the stimulation of 10ng/mL TNF-α, the expression of CTSK and TRAP was markedly upregulated in PBMCs. Meanwhile, the results of Western blot and real-time RT-PCR showed that the expression of cytoplasmic phosphorylated inhibitor of NF-κB α (p-IκBα) and nuclear p65 in SHED were significantly higher than that without TNF-α stimulation after 10 ng/mL TNF-α treatment.

CONCLUSIONS

TNF-α regulates the ability of SHED to promote osteoclastogenesis through NF-κB signal pathways.

摘要

目的

研究肿瘤坏死因子-α（TNF-α）对人脱落乳牙干细胞（SHED）促进破骨细胞生成能力的影响。

方法

从乳牙中获取SHED并进行分离、纯化及体外培养。建立SHED与破骨细胞前体外周血单个核细胞（PBMCs）的间接共培养体系。用TNF-α（0、5、10、50、100 ng/mL）处理后，通过实时逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测PBMCs中破骨细胞相关基因及SHED中核因子κB（NF-κB）的表达。采用SPSS 19.0软件包进行统计分析。

结果

在10 ng/mL TNF-α刺激下，PBMCs中组织蛋白酶K（CTSK）和抗酒石酸酸性磷酸酶（TRAP）的表达明显上调。同时，蛋白质免疫印迹法和实时RT-PCR结果显示，10 ng/mL TNF-α处理后，SHED中细胞质磷酸化核因子κB抑制蛋白α（p-IκBα）和细胞核p65的表达显著高于未用TNF-α刺激时。