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

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。

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

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.