Zhang Jinlong, Lian Min, Cao Peipei, Bao Guofeng, Xu Guanhua, Sun Yuyu, Wang Lingling, Chen Jiajia, Wang Yi, Feng Guijuan, Cui Zhiming
Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China.
Department of Stomatology, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People's Republic of China.
Neurochem Res. 2017 Apr;42(4):1015-1025. doi: 10.1007/s11064-016-2134-3. Epub 2016 Dec 22.
Dental pulp stem cells (DPSCs) were the most widely used seed cells in the field of neural regeneration and bone tissue engineering, due to their easily isolation, lack of ethical controversy, low immunogenicity and low rates of transplantation rejection. The purpose of this study was to investigate the role of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) on neural differentiation of DPSCs in vitro. DPSCs were cultured in neural differentiation medium containing NGF and bFGF alone or combination for 7 days. Then neural genes and protein markers were analyzed using western blot and RT-PCR. Our study revealed that bFGF and NGF increased neural differentiation of DPSCs synergistically, compared with bFGF and NGF alone. The levels of Nestin, MAP-2, βIII-tubulin and GFAP were the most highest in the DPSCs + bFGF + NGF group. Our results suggested that bFGF and NGF signifiantly up-regulated the levels of Sirt1. After treatment with Sirt1 inhibitor, western blot, RT-PCR and immunofluorescence staining showed that neural genes and protein markers had markedly decreased. Additionally, the ERK and AKT signaling pathway played a key role in the neural differentiation of DPSCs stimulated with bFGF + NGF. These results suggested that manipulation of the ERK and AKT signaling pathway may be associated with the differentiation of bFGF and NGF treated DPSCs. Our date provided theoretical basis for DPSCs to treat neurological diseases and repair neuronal damage.
牙髓干细胞（DPSCs）因其易于分离、不存在伦理争议、免疫原性低和移植排斥率低，成为神经再生和骨组织工程领域应用最广泛的种子细胞。本研究旨在探讨碱性成纤维细胞生长因子（bFGF）和神经生长因子（NGF）在体外对DPSCs神经分化的作用。将DPSCs单独或联合添加NGF和bFGF培养于神经分化培养基中7天。然后采用蛋白质免疫印迹法和逆转录-聚合酶链反应（RT-PCR）分析神经基因和蛋白质标志物。我们的研究表明，与单独使用bFGF和NGF相比，bFGF和NGF协同促进DPSCs的神经分化。在DPSCs+bFGF+NGF组中，巢蛋白（Nestin）、微管相关蛋白2（MAP-2）、βIII-微管蛋白（βIII-tubulin）和胶质纤维酸性蛋白（GFAP）的水平最高。我们的结果表明，bFGF和NGF显著上调了沉默信息调节因子1（Sirt1）的水平。用Sirt1抑制剂处理后，蛋白质免疫印迹法、RT-PCR和免疫荧光染色显示神经基因和蛋白质标志物明显减少。此外，细胞外信号调节激酶（ERK）和蛋白激酶B（AKT）信号通路在bFGF+NGF刺激的DPSCs神经分化中起关键作用。这些结果表明，ERK和AKT信号通路的调控可能与bFGF和NGF处理的DPSCs的分化有关。我们的数据为DPSCs治疗神经疾病和修复神经元损伤提供了理论依据。
