Gyetvai Georgina, Hughes Trisha, Wedmore Florence, Roe Cieron, Heikal Lamia, Ghezzi Pietro, Mengozzi Manuela
Department of Clinical and Experimental Medicine, Brighton and Sussex Medical School, Brighton, United Kingdom.
Front Immunol. 2017 Oct 26;8:1394. doi: 10.3389/fimmu.2017.01394. eCollection 2017.
Several studies have shown that erythropoietin (EPO) has neuroprotective or neuroreparative actions on diseases of the nervous system and that improves oligodendrocyte (OL) differentiation and myelination and . This study aims at investigating the early molecular mechanisms for the pro-myelinating action of EPO at the gene expression level. For this purpose, we used a differentiating OL precursor cell line, rat central glia-4 cells. Cells were differentiated or not, and then treated with EPO for 1 or 20 h. RNA was extracted and changes in the gene expression profile were assessed using microarray analysis. Experiments were performed in biological replicates of = 4. Differentiation alone changed the expression of 11% of transcripts (2,663 out of 24,272), representing 2,436 genes, half of which were upregulated and half downregulated. At 20 h of treatment, EPO significantly affected the expression of 99 genes that were already regulated by differentiation and of 150 genes that were not influenced by differentiation alone. Analysis of the transcripts most upregulated by EPO identified several genes involved in lipid transport (e.g., ) and lipid metabolism () along with and , growth factors known for their pro-myelinating action. All these genes were only induced by EPO and not by differentiation alone, except for which was highly induced by differentiation and augmented by EPO. Results were validated by quantitative PCR. These findings suggest that EPO might increase remyelination by inducing insulin-like growth factors and increasing lipid metabolism.
多项研究表明，促红细胞生成素（EPO）对神经系统疾病具有神经保护或神经修复作用，可促进少突胶质细胞（OL）分化和髓鞘形成。本研究旨在从基因表达水平探究EPO促髓鞘形成作用的早期分子机制。为此，我们使用了一种正在分化的OL前体细胞系——大鼠中枢神经胶质-4细胞。细胞进行或不进行分化处理，然后用EPO处理1或20小时。提取RNA，并使用微阵列分析评估基因表达谱的变化。实验进行了4次生物学重复。仅分化就改变了11%的转录本（24272个中的2663个）的表达，代表2436个基因，其中一半上调，一半下调。在处理20小时时，EPO显著影响了99个已受分化调控的基因以及150个仅分化未影响的基因的表达。对EPO上调最明显的转录本分析发现，有几个基因参与脂质转运（如）和脂质代谢（），以及和这些以其促髓鞘形成作用而闻名的生长因子。除了在分化时高度诱导且EPO使其增强外，所有这些基因仅由EPO诱导而非仅由分化诱导。结果通过定量PCR得到验证。这些发现表明，EPO可能通过诱导胰岛素样生长因子和增加脂质代谢来增加髓鞘再生。
