TNF-α对人脱落乳牙牙髓干细胞促破骨细胞形成能力的影响

doi:10.19439/j.sjos.2018.05.001

上海口腔医学 ›› 2018, Vol. 27 ›› Issue (5): 449-454.doi: 10.19439/j.sjos.2018.05.001

TNF-α对人脱落乳牙牙髓干细胞促破骨细胞形成能力的影响

王宇琛¹, 王辰², 刘娜¹, 蔡川¹, 李伟¹, 徐璐璐¹

1.中国人民解放军总医院口腔正畸科,北京 100853;
2.中国人民解放军第309医院口腔科,北京 100091

收稿日期:2018-02-05 出版日期:2018-10-25 发布日期:2018-11-05
通讯作者: 徐璐璐, E-mail：xululu1977@163.com
作者简介:王宇琛（1987-）,男,硕士,住院医师,E-mail：andychen19871014 @126.com
基金资助:
科技部国际科技合作项目（2015-TSYS-2028）; 国家自然科学基金（81701018,8150030603）; 解放军总医院临床科研扶持基金（2017FC-TSYS-3013）

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

WANG Yu-chen¹, WANG Chen², LIU Na¹, CAI Chuan¹, LI Wei¹, XU Lu-lu¹

1.Department of Orthodontics, General Hospital of Chinese PLA. Beijing 100853;
2.Department of Stomatology, 309th Hospital of Chinese PLA. Beijing 100091, China

Received:2018-02-05 Online:2018-10-25 Published:2018-11-05

摘要/Abstract

摘要： 目的: 探讨肿瘤坏死因子α（tumor necrosis factor -α,TNF-α）对人脱落乳牙牙髓干细胞（stem cells from human exfoliated deciduous teeth,SHED）促破骨细胞形成能力的影响。方法: 通过酶消化法体外分离、培养处于生理性根吸收时期的自然脱落乳牙牙髓干细胞;建立SHED与破骨前体细胞外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）的间接共培养模型,在破骨诱导培养液中加入0、5、10、50、100 ng/mL不同浓度的TNF-α,通过实时定量RT-PCR和Western 免疫印迹检测破骨相关基因及核转录因子κB（nuclear factor-κB,NF-κB）信号通路相关基因的表达。采用SPSS 19.0软件包对数据进行统计学分析。结果: 在10 ng/mL的TNF-α刺激下,PBMCs中破骨相关蛋白CTSK和 TRAP 的表达水平均显著增高。Western免疫印迹和实时定量RT-PCR检测结果显示,SHED胞质内 p-IκBα 和胞核内 p65 蛋白、基因的表达水平在10 ng/mL的TNF-α刺激下显著高于无TNF-α刺激组。结论: 炎症细胞因子TNF-α通过NF-κB信号通路对SHED促破骨细胞形成能力具有调控作用。

关键词: 人脱落乳牙牙髓干细胞, 肿瘤坏死因子α, 核转录因子κB信号通路, 破骨细胞分化

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.

Key words: Stem cells from human exfoliated deciduous teeth, Tumor necrosis factor-α, Nuclear factor-κB signal pathways, Osteoclastic differentiation

中图分类号:

Q254

王宇琛, 王辰, 刘娜, 蔡川, 李伟, 徐璐璐. TNF-α对人脱落乳牙牙髓干细胞促破骨细胞形成能力的影响[J]. 上海口腔医学, 2018, 27(5): 449-454.

WANG Yu-chen, WANG Chen, LIU Na, CAI Chuan, LI Wei, XU Lu-lu. Effect of TNF-α on the ability of stem cells from human exfoliated deciduous teeth to promote osteoclastogenesis[J]. Shanghai Journal of Stomatology, 2018, 27(5): 449-454.

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TNF-α对人脱落乳牙牙髓干细胞促破骨细胞形成能力的影响

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

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