The role of Wnt7B in the mediation of dentinogenesis via the ERK1/2 pathway
The role of Wnt7B in the mediation of dentinogenesis via the ERK1/2 pathway
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Wnt7B 通过 ERK1/2 途径介导牙本质发生的作用
DOI：
10.1016/j.archoralbio.2019.05.009
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发表时间：
2019-08-01
影响因子：
3
通讯作者：
Ye, Ling
中科院分区：
文献类型：
--
作者：
Chen, Dian;Yu, Fanyuan;Ye, Ling
目的：本研究旨在探讨Wnt 7 b在小鼠牙本质形成中的作用。设计：收集C57 BL/6小鼠牙胚，采用免疫组织化学方法检测Wnt 7 b在不同发育阶段牙胚中的表达。通过Micro-CT和HE染色分析Dmp 1-cre; ROSA 26-Wnt 7 b转基因小鼠和ROSA 26-Wnt 7 b同窝小鼠的下颌骨形态。电镜下观察牙本质超微结构。原代培养的小鼠牙乳头细胞（MDPCs）和成牙本质细胞系（A11）用腺病毒感染以过表达Wnt 7 b。CCK-8和流式细胞仪检测细胞增殖和凋亡。通过茜素红染色和qPCR检测成骨基因表达来评估MDPCs和A11的成骨分化。通过使用蛋白质印迹分析来测量信号传导途径的活化。结果：Wnt 7 b主要表达于钟状期后的成牙本质细胞层，在成牙本质细胞层中表达的Wnt 7 b主要表达于钟状期后的成牙本质细胞层。在转基因小鼠中，Wnt 7 b在牙齿间充质中过表达，具有较薄的前牙本质层和较厚的管间牙本质。转基因小鼠牙本质的显微硬度值和Ca/Pi比值均高于对照组。Wnt 7 b促进MDPCs和A11的增殖和矿化。Ad-Wnt 7 b感染A11细胞后，p-ERK 1/2蛋白水平明显升高。结论：Wnt 7 b促进牙本质细胞的增殖和分化，其机制可能与ERK 1/2信号通路有关。
Objectives: This study investigates the role of Wnt7b in mouse dentin formation.Design: C57BL/6 mouse tooth germs at different developmental stages were collected to measure the expression of Wnt7b by immunohistochemical staining. The morphology of mandibles of Dmp1-cre;ROSA26-Wnt7b transgenic mice and ROSA26-Wnt7b littermates was analyzed by Micro-CT and HE staining. The ultramicrostructure of dentin was scanned with an electron microscope. Primary mouse dental papillae cells (MDPCs) and odontoblastic cell line (A11) were cultured and infected with adenovirus to overexpress Wnt7b. Cell proliferation and cell apoptosis were evaluated using CCK-8 and flow cytometry. Osteogenic differentiation of MDPCs and A11 was assessed by Alizarin red staining, and qPCR detection of osteogenic gene expression. The activation of signaling pathways was measured by the use of western blot analysis. The ERK1/2 inhibitor was used to test the effect of Wnt7b regulated cell differentiation.Results: Wnt7b was expressed principally in the mouse odontoblast layer after the early bell stage. In transgenic mice, Wnt7b was over-expressed in tooth mesenchyme, with a thinner predentin layer and thicker intertubular dentin. Both the micro-hardness value and the Ca/Pi ratio of dentin of transgenic mice were higher. Wnt7b promoted proliferation and mineralization of MDPCs and A11. The protein level of p-ERK1/2 was found to be higher in A11 infected with Ad-Wnt7b. The ERK signaling pathway inhibitor partly rescued the Wnt7b-induced differentiation of A11.Conclusions: Wnt7b enhances dentinogenesis by increasing the proliferation and differentiation of dental mesenchymal cells partly through ERK1/2 pathway.