Departamento de Bioquímica, Centro de Estudos em Estresse Oxidativo, Universidade Federal do Rio Grande do Sul-UFRGS, Rua Ramiro Barcelos 2600, Cep 90035-003, Porto Alegre, Rio Grande do Sul, Brazil.
Departamento de Bioquímica, Instituto de Medicina Tropical, Universidade Federal do Rio Grande do Norte-UFRN, Avenida Senador Salgado Filho 3000, Cep 59078-900, Natal, Rio Grande do Norte, Brazil.
Mol Neurobiol. 2016 Jan;53(1):423-435. doi: 10.1007/s12035-014-8998-9. Epub 2014 Dec 3.
SH-SY5Y cells, a neuroblastoma cell line that is a well-established model system to study the initial phases of neuronal differentiation, have been used in studies to elucidate the mechanisms of neuronal differentiation. In the present study, we investigated alterations of gene expression in SH-SY5Y cells during neuronal differentiation mediated by retinoic acid (RA) treatment. We evaluated important pathways involving nuclear factor kappa B (NF-κB), nuclear E2-related factor 2 (Nrf2), glycolytic, and p53 during neuronal differentiation. We also investigated the involvement of reactive oxygen species (ROS) in modulating the gene expression profile of those pathways by antioxidant co-treatment with Trolox®, a hydrophilic analogue of α-tocopherol. We found that RA treatment increases levels of gene expression of NF-κB, glycolytic, and antioxidant pathway genes during neuronal differentiation of SH-SY5Y cells. We also found that ROS production induced by RA treatment in SH-SY5Y cells is involved in gene expression profile alterations, chiefly in NF-κB, and glycolytic pathways. Antioxidant co-treatment with Trolox® reversed the effects mediated by RA NF-κB, and glycolytic pathways gene expression. Interestingly, co-treatment with Trolox® did not reverse the effects in antioxidant gene expression mediated by RA in SH-SY5Y. To confirm neuronal differentiation, we quantified endogenous levels of tyrosine hydroxylase, a recognized marker of neuronal differentiation. Our data suggest that during neuronal differentiation mediated by RA, changes in profile gene expression of important pathways occur. These alterations are in part mediated by ROS production. Therefore, our results reinforce the importance in understanding the mechanism by which RA induces neuronal differentiation in SH-SY5Y cells, principally due this model being commonly used as a neuronal cell model in studies of neuronal pathologies.
SH-SY5Y 细胞是一种神经母细胞瘤细胞系，是研究神经元分化初始阶段的成熟模型系统，已被用于阐明神经元分化机制的研究。在本研究中，我们研究了维甲酸（RA）处理介导的 SH-SY5Y 细胞神经元分化过程中基因表达的变化。我们评估了核因子 kappa B（NF-κB）、核 E2 相关因子 2（Nrf2）、糖酵解和 p53 等重要途径在神经元分化过程中的作用。我们还研究了抗氧化剂 Trolox®（α-生育酚的亲水性类似物）共处理通过调节这些途径的基因表达谱来调节活性氧（ROS）的参与。我们发现 RA 处理增加了 NF-κB、糖酵解和抗氧化途径基因在 SH-SY5Y 细胞神经元分化过程中的基因表达水平。我们还发现 RA 处理在 SH-SY5Y 细胞中诱导的 ROS 产生参与了基因表达谱的改变，主要影响 NF-κB 和糖酵解途径。抗氧化剂 Trolox®共处理逆转了 RA 介导的 NF-κB 和糖酵解途径基因表达的作用。有趣的是，Trolox®共处理并没有逆转 RA 介导的抗氧化基因表达的作用。为了确认神经元分化，我们量化了内源性酪氨酸羟化酶的水平，这是神经元分化的公认标志物。我们的数据表明，在 RA 介导的神经元分化过程中，重要途径的基因表达谱发生了变化。这些改变部分是由 ROS 产生介导的。因此，我们的结果强调了理解 RA 在 SH-SY5Y 细胞中诱导神经元分化的机制的重要性，主要是因为该模型通常被用作神经元病理学研究中的神经元细胞模型。
