EphrinB2 signalling modulates the neural differentiation of human dental pulp stem cells
EphrinB2 signalling modulates the neural differentiation of human dental pulp stem cells
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EphrinB2信号调节人牙髓干细胞的神经分化
DOI：
10.3892/br.2018.1108
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发表时间：
2018-08-01
影响因子：
2.3
通讯作者：
Zhang, Chengfei
中科院分区：
文献类型：
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作者：
Heng, Boon Chin;Gong, Ting;Zhang, Chengfei
牙髓干细胞(DPSCs)起源于胚胎神经嵴，具有神经分化潜能。本研究探讨了正向和反向EPhinB2信号通路在DPSC神经发生中的作用。在神经诱导培养7d后，重组EphainB2-Fc蛋白作用于DPSCs后，神经标志物：β-微管蛋白、神经细胞黏附分子(NCAM)、巢蛋白、神经原蛋白2(NGN2)、神经丝介质多肽和Musashi1的表达显著下调。免疫细胞化学显示，经EphainB2-Fc处理的DPSCs形态更圆，突起较少，βIII-微管蛋白和Ngn2蛋白表达减少。用EphB4受体特异性多肽抑制剂处理DPSCs可显著上调神经标记物微管相关蛋白2、Musashi1、Ngn2和神经元特异性烯醇化酶的表达，而用EphB2受体特异性多肽抑制剂处理DPSCs对神经发生的影响不大。在DPSCs中转基因表达EPhinB2导致Musashi1和NCAM基因表达显著上调，而重组EphB4-Fc蛋白处理DPSCs仅显著上调Musashi1基因表达。因此，可以得出这样的结论：刺激前向信号转导EPhinB2-EphB4显著抑制DPSCs的神经发生，而用EphB4特异性的多肽抑制剂抑制这一前向信号通路则促进神经发生。同时，刺激反向EphB4-EPhinB2信号仅略微增强了DPSCs的神经分化。本研究结果提示，在DPSCs的神经组织工程中，有可能使用EPhinB2正向信号转导的多肽或小分子抑制剂。
Dental pulp stem cells (DPSCs) originate from the embryonic neural crest and have neurogenic potential. The present study investigated the roles of the forward and reverse EphrinB2 signalling pathways during DPSC neurogenesis. Treatment of DPSCs with recombinant EphrinB2-Fc protein over 7 days in a neural induction culture resulted in significant downregulation of the following neural markers: beta III-Tubulin, neural cell adhesion molecule (NCAM), nestin, neurogenin 2 (NGN2), neurofilament medium polypeptide and Musashi1. Immunocytochemistry revealed that EphrinB2-Fc-treated DPSCs exhibited more rounded morphologies with fewer neurite outgrowths as well as reduced protein expression of beta III-tubulin and NGN2. Treatment of DPSCs with a peptide inhibitor specific to the EphB4 receptor significantly upregulated expression of the neural markers microtubule-associated protein 2, Musashi1, NGN2 and neuron-specific enolase, whereas treatment with a peptide inhibitor specific to the EphB2 receptor exerted negligible effects on neurogenesis. Transgenic expression of EphrinB2 in DPSCs resulted in significant upregulation of Musashi1 and NCAM gene expression, while treatment of DPSCs with recombinant EphB4-Fc protein led to significant upregulation of only Musashi1. Thus, it may be concluded that stimulation of forward EphrinB2-EphB4 signalling markedly inhibited neurogenesis in DPSCs, whereas suppression of this forward signalling pathway with peptide inhibitor specific to EphB4 promoted neurogenesis. Meanwhile, stimulation of reverse EphB4-EphrinB2 signalling only marginally enhanced the neural differentiation of DPSCs. The present findings indicate the potential application of peptide or small molecule inhibitors of EphrinB2 forward signalling in neural tissue engineering with DPSCs.