miR-143 通过靶向 TNF-α 抑制 NF-κB 信号通路来抑制牙髓干细胞的成骨分化。
miR-143 suppresses the osteogenic differentiation of dental pulp stem cells by inactivation of NF-κB signaling pathway via targeting TNF-α.
机构信息
Department of Prosthodontics, The First Affiliated Hospital & School of Stomatology, Zhengzhou University, No. 40 Daxue Road, Zhengzhou, 450052, China.
Department of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
出版信息
Arch Oral Biol. 2018 Mar;87:172-179. doi: 10.1016/j.archoralbio.2017.12.031. Epub 2017 Dec 30.
BACKGROUND
Dental pulp stem cells (DPSCs) are multipotent and play an important role in repairing damaged and/or defective dentinogenesis/osteogenesis. Recent studies have documented the implication of miR-143 in osteogenic differentiation of DPSCs. Nevertheless, the detailed mechanisms of miR-143 involved in the osteogenic differentiation of DPSCs remain to be further elaborated.
METHODS
Isolated DPSCs were incubated with osteogenic differentiation medium to induce osteogenic differentiation. qRT-PCR and western blot were performed to determine the expressions of miR-143 and tumor necrosis factor α (TNF-α). Luciferase reporter assay was used to confirm whether TNF-α was a target of miR-143. Osteogenic differentiation of DPSCs was evaluated by alkaline phosphatase (ALP) activity assay, ALP staining, and western blot analyses of osteogenic-markers including bone morphogenetic protein 2 (BMP2), ALP, runt-related transcription factor 2 (RUNX2) and collagen type I (COLI).
RESULTS
miR-143 was downregulated and TNF-α was upregulated during osteogenic differentiation of DPSCs. miR-143 posttranscriptionally regulated TNF-α expression in DPSCs by binding to its 3'UTR. miR-143 overexpression suppressed osteogenic differentiation of DPSCs, as demonstrated by the decrease of ALP activity, ALP positive cell ratio, as well as BMP2, ALP, RUNX2, and COLI expressions. Moreover, miR-143 reversed TNF-α-induced osteogenic differentiation of DPSCs. Finally, the osteogenic differentiation of DPSCs induced by miR-143 inhibitor was attenuated following inactivation of nuclear factor kappa B (NF-κB) signaling pathway.
CONCLUSION
miR-143 suppressed the osteogenic differentiation of DPSCs by blockade of NF-κB signaling pathway via targeting TNF-α.
背景
牙髓干细胞（DPSCs）具有多能性，在修复受损和/或有缺陷的牙本质/骨生成中起着重要作用。最近的研究记录了 miR-143 在 DPSCs 成骨分化中的作用。然而，miR-143 参与 DPSCs 成骨分化的详细机制仍有待进一步阐述。
方法
分离的 DPSCs 用成骨分化培养基孵育以诱导成骨分化。通过 qRT-PCR 和 Western blot 测定 miR-143 和肿瘤坏死因子α（TNF-α）的表达。荧光素酶报告实验用于确认 TNF-α是否是 miR-143 的靶标。通过碱性磷酸酶（ALP）活性测定、ALP 染色和骨形成蛋白 2（BMP2）、ALP、 runt 相关转录因子 2（RUNX2）和胶原 I（COLI）等成骨标志物的 Western blot 分析评估 DPSCs 的成骨分化。
结果
在 DPSCs 的成骨分化过程中，miR-143 下调，TNF-α上调。miR-143 通过与 3'UTR 结合，在后转录水平调节 DPSCs 中的 TNF-α表达。miR-143 过表达抑制 DPSCs 的成骨分化，表现为 ALP 活性、ALP 阳性细胞比例以及 BMP2、ALP、RUNX2 和 COLI 表达降低。此外，miR-143 逆转了 TNF-α诱导的 DPSCs 成骨分化。最后，miR-143 抑制剂诱导的 DPSCs 成骨分化在 NF-κB 信号通路失活后减弱。
结论
miR-143 通过靶向 TNF-α阻断 NF-κB 信号通路抑制 DPSCs 的成骨分化。
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