上海口腔医学

上海口腔医学 ›› 2024, Vol. 33 ›› Issue (2): 135-140.doi: 10.19439/j.sjos.2024.02.005

• 论著 • 上一篇    下一篇

TNF-α通过ERK1/2-Runx2信号通路调控SHED成骨分化能力的实验研究

王静1,2, 徐娜1,2, 任慧迪1,2   

  1. 1.天津市口腔医院 儿童口腔科,南开大学医学院,天津 300041;
    2.天津市口腔功能重建重点实验室, 天津 300041
  • 收稿日期:2023-12-25 修回日期:2024-01-24 出版日期:2024-04-25 发布日期:2024-05-14
  • 通讯作者: 王静,E-mail：wangjing11syea@163.com
  • 作者简介:王静（1992-）,女,硕士,主治医师
  • 基金资助:
    天津市口腔医院重点学科科研项目（2022EK08）

TNF-α regulated SHED osteogenic differentiation through ERK1/2-Runx2 signaling pathway

WANG Jing1,2, XU Na1,2, REN Hui-di1,2   

  1. 1. Department of Pediatric Dentistry,Tianjin Stomatological Hospital; School of Medicine, Nankai University. Tianjin 300041;
    2. Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction. Tianjin 300041, China
  • Received:2023-12-25 Revised:2024-01-24 Online:2024-04-25 Published:2024-05-14

PDF (PC)

44

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）骨分化能力的影响,分析ERK1/2-Runx2信号通路在该调控过程中的变化。方法: 从6~8 岁健康儿童正常乳恒牙替换即将脱落的乳切牙中分离和培养 SHED,取第三代细胞,分为对照组（成骨诱导剂培养）、观察组（成骨诱导剂和TNF-α共培养）和激动剂组（成骨诱导剂、TNF-α和ERK通路激动剂共培养）。采用茜素红染色评价成骨分化功能,采用 Western 印迹检测SHED 细胞中 Osterix、OPN、ERK1/2、pERK1/2和Runx2 的蛋白表达水平,应用 qRT-PCR 检测Osterix、OPN、ERK1/2、pERK1/2和Runx2 mRNA的表达。采用SPSS 26.0软件包对数据进行统计学分析。结果: 3组细胞成骨分化能力比较结果显示,3组细胞中均可见红棕色矿化结节。3组组间相比,对照组矿化结节最多,激动剂组次之,观察组最少。与对照组相比,观察组和激动剂组的Osterix、OPN 蛋白和mRNA表达水平显著下降,而激动剂组Osterix、OPN 蛋白和mRNA表达水平显著高于观察组;3组细胞的ERK1/2蛋白和mRNA表达水平无显著差异,而观察组和激动剂组pERK1 /2和Runx2的蛋白和mRNA表达水平显著高于对照组,激动剂组的蛋白及mRNA表达水平显著高于观察组。结论: TNF-α 对SHED成骨分化具有抑制作用,该作用可能与抑制ERK1/2-Runx2信号通路有关。

关键词: 肿瘤坏死因子α, 人脱落乳牙牙髓干细胞, 成骨, 分化, ERK1/2-Runx2信号通路

Abstract: PURPOSE: To investigate the effect of TNF-α on osteogenic differentiation of stem cells from human exfoliated deciduous teeth (SHED), and to analyze the changes of ERK1/2-Runx2 signaling pathway in the regulation process. METHODS: SHED cells were isolated and cultured from normal deciduous permanent teeth of healthy children aged 6-8 years old, and the third passage of SHED cells were taken and divided into control group (osteogenic inducer culture), observation group (osteogenic inducer and TNF-α co-culture) and agonist group (osteogenic inducer, TNF-α and ERK pathway agonist co-culture). The osteogenic differentiation was determined by alizarin red staining. The protein expression levels of Osterix, OPN, ERK1/2, pERK1/2 and Runx2 in SHED cells were determined by Western blot. The expressions of Osterix, OPN, ERK1/2, pERK1/2 and Runx2 mRNA were detected by qRT-PCR. Statistical analysis was performed with SPSS 26.0 software package. RESULTS: Comparison of osteogenic differentiation ability of the three groups of cells showed that red-brown mineralized nodules were observed in the three groups of cells. Compared among the three groups, the control group had the most mineralized nodules, followed by the activation group, and the observation group had the least mineralized nodules. Compared with the control group, the expression levels of Osterix and OPN protein and mRNA in the observation group and the agonist group were significantly decreased, while the expression levels of Osterix and OPN protein and mRNA in the agonist group were significantly higher than those in the observation group. There was no significant difference in the expression levels of ERK1/2 protein and mRNA among the three groups, while the expression levels of pERK1/2 and Runx2 protein and mRNA in the observation group and the agonist group were significantly higher than those in the control group, and the expression levels of pERK1/2 and Runx2 protein and mRNA in the agonist group were significantly higher than those in the observation group. CONCLUSIONS: TNF-α can inhibit osteogenic differentiation of SHED cells, which may be related to the inhibition of ERK1/2-Runx2 signaling pathway.

Key words: Tumor necrosis factor α, Human deciduous tooth pulp stem cells, Osteogenesis, Differentiation, ERK1/2-Runx2 signaling pathway

中图分类号: