Magnesium-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells by activating ERK/BMP2/Smads signaling
Magnesium-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells by activating ERK/BMP2/Smads signaling
复制标题
富镁微环境通过激活ERK/BMP2/Smads信号促进人牙髓干细胞的牙源性分化
DOI：
10.1186/s13287-019-1493-5
复制
发表时间：
2019-12-10
影响因子：
7.5
通讯作者：
Zheng, Jianmao
中科院分区：
文献类型：
--
作者：
Kong, Yuanyuan;Hu, Xiaoli;Zheng, Jianmao
背景资料：富镁微环境促进人牙髓干细胞向牙源性分化，但其调控机制尚不清楚。本工作的目的是评估镁的功能在上述过程中，并探讨相关的信号pathway.Methods：DPSCs进行培养在牙源性培养基中添加0，1，5，或10 mM的MgCl 2。使用Mag-Fluo-4-AM流式细胞术评估DPSC中的细胞内Mg 2+水平。TRPM 7抑制剂2-氨基乙氧基二苯基硼酸酯（2-APB）可抑制Mg ~（2+）的内流。进行RNA测序以评估与高细胞外Mg 2+相关的牙源性分化过程中DPSC的转录组改变。进行KEGG通路分析以确定与检索到的差异表达基因（DEG）相关的通路。结果：富镁微环境通过增加细胞内Mg 2+浓度促进牙髓干细胞向牙源性分化。一致地，高细胞外Mg 2+对DPSC牙源性分化的积极作用被2-APB阻断，从而减少了Mg 2+的进入。RNA测序鉴定了734个与高细胞外Mg 2+存在下DPSC牙向分化相关的DEG。这些DEG参与了许多级联反应，如MAPK和TGF-β途径。高浓度Mg 2+处理后，DPSCs中ERK和BMP 2/Smads通路均被激活。U 0126抑制ERK信号通路可减弱高浓度Mg 2+对DPSCs矿化和牙源性分化的影响。结论：富镁微环境通过激活ERK/BMP 2/Smads信号通路促进牙髓干细胞向牙源性分化。本研究表明，富镁微环境可作为牙髓再生的一种新策略。
Background: Magnesium (Mg2+)-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells (DPSCs), but the regulatory mechanisms remain undefined. The aim of this work was to assess magnesium's function in the above process and to explore the associated signaling pathway.Methods: DPSCs underwent culture in odontogenic medium with the addition of 0, 1, 5, or 10 mM MgCl2. Intracellular Mg2+ levels in DPSCs were evaluated flow cytometrically using Mag-Fluo-4-AM. Mg2+-entry was inhibited by TRPM7 inhibitor 2-aminoethoxydiphenyl borate (2-APB). RNA-Sequencing was carried out for assessing transcriptome alterations in DPSCs during odontogenic differentiation associated with high extracellular Mg2+. KEGG pathway analysis was performed to determine pathways related to the retrieved differentially expressed genes (DEGs). Immunoblot was performed for assessing magnesium's role and exploring ERK/BMP2/Smads signaling.Results: Mg2+-enriched microenvironment promoted odontogenic differentiation in DPSCs via intracellular Mg2+ increase. Consistently, the positive effect of high extracellular Mg2+ on odontogenic differentiation in DPSCs was blocked by 2-APB, which reduced Mg2+ entry. RNA-sequencing identified 734 DEGs related to odontogenic differentiation in DPSCs in the presence of high extracellular Mg2+. These DEGs participated in many cascades such as MAPK and TGF-beta pathways. Consistently, ERK and BMP2/Smads pathways were activated in DPSCs treated with high extracellular Mg2+. In agreement, ERK signaling inhibition by U0126 blunted the effect of high extracellular Mg2+ on mineralization and odontogenic differentiation in DPSCs. Interestingly, BMP2, BMPR1, and phosphorylated Smad1/5/9 were significantly decreased by U0126, indicating that BMP2/Smads acted as downstream of ERK.Conclusions: Mg2+-enriched microenvironment promotes odontogenic differentiation in DPSCs by activating ERK/BMP2/Smads signaling via intracellular Mg2+ increase. This study revealed that Mg2+-enriched microenvironment could be used as a new strategy for dental pulp regeneration.