Geng Ya-Wei, Zhang Zhen, Liu Ming-Yue, Hu Wei-Ping
Department of Prosthodontics, 2nd Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150086, China.
Oral and Maxillofacial Surgery, 2nd Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150086, China.
Cell Biol Int. 2017 Dec;41(12):1391-1398. doi: 10.1002/cbin.10835. Epub 2017 Aug 29.
Dental pulp stem cells (DPSCs) have been proposed as a promising source of stem cells in nerve regeneration due to their close embryonic origin and ease of harvest. Resveratrol (RSV) is a natural polyphenolic and possesses many biological functions such as anti-inflammatory activity and protection against atherosclerosis and neuroprotective activities. There is increasing evidence showing that RSV plays a pivotal role in neuron protection and neuronal differentiation. In this study, we isolated DPSCs from impacted third molars and investigated whether RSV induces neuronal differentiation of DPSCs. To avoid loss of DPSCs multipotency, all the experiments were conducted on cells at early passages. RT-PCR results showed that RSV-treated DPSCs (RSV-DPSCs) significantly increased the expression of the neuroprogenitor marker Nestin. When RSV-DPSCs were differentiated with neuronal induction media (RSV-dDPSCs), they showed a cell morphology similar to neurons. The expression of neuronal-specific marker genes Nestin, Musashi, and NF-M in RSV-dDPSCs was significantly increased. Immunocytochemical staining and Western blot analysis showed that the expression of neuronal marker proteins, Nestin, and NF-M, was significantly increased in RSV-dDPSCs. Therefore, we have shown that RSV treatment, along with the use of neuronal induction media, effectively promotes neuronal cell differentiation of DPSCs.
牙髓干细胞（DPSCs）因其与胚胎起源相近且易于获取，已被认为是神经再生中一种很有前景的干细胞来源。白藜芦醇（RSV）是一种天然多酚，具有多种生物学功能，如抗炎活性、抗动脉粥样硬化作用以及神经保护活性。越来越多的证据表明，RSV在神经元保护和神经元分化中起关键作用。在本研究中，我们从阻生第三磨牙中分离出DPSCs，并研究RSV是否能诱导DPSCs向神经元分化。为避免DPSCs多能性丧失，所有实验均在早期传代的细胞上进行。逆转录聚合酶链反应（RT-PCR）结果显示，经RSV处理的DPSCs（RSV-DPSCs）显著增加了神经祖细胞标志物巢蛋白（Nestin）的表达。当RSV-DPSCs用神经元诱导培养基进行分化时（RSV-dDPSCs），它们呈现出与神经元相似的细胞形态。RSV-dDPSCs中神经元特异性标志物基因Nestin、Musashi和神经丝蛋白M（NF-M）的表达显著增加。免疫细胞化学染色和蛋白质印迹分析表明，RSV-dDPSCs中神经元标志物蛋白Nestin和NF-M的表达显著增加。因此，我们已经表明，RSV处理以及使用神经元诱导培养基能有效促进DPSCs向神经元细胞分化。
