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目的探讨睫状神经营养因子(CNTF)对受损视神经胶质细胞去分化的作用及机制。方法将65只SD大鼠随机分为对照组、损伤组和CNTF组,制备视神经损伤及损伤后添加CNTF的动物模型。术后7、14 d取术侧视神经损伤远侧段,分别用基因芯片检测其基因表达谱,实时荧光定量PCR、免疫组织化学、HE染色等方法检测相关基因、蛋白表达和细胞数量的变化。结果术后7 d,与损伤组相比,CNTF组筛选出608条表达上调和417条表达下调的差异基因,包括染色质构型、转录调节、神经干细胞、神经分化和发育、增殖凋亡、离子通道、受体、信号转导等相关基因。实时荧光定量PCR结果验证了基因芯片结果的可靠性。CNTF组视神经损伤远侧段细胞数量增多,远侧段Nestin、胶质纤维酸性蛋白(GFAP)、髓鞘碱性蛋白(MBP)、细胞外信号调节激酶1/2(Erk1/2)和近侧段神经丝(NF)阳性物质增多。结论外源性CNTF通过调节受损视神经大胶质细胞的去分化相关基因及蛋白的表达,促进了胶质细胞的去分化和轴突再生。
Objective To investigate the effect and mechanism of ciliary neurotrophic factor (CNTF) on the dedifferentiation of impaired optic neuroglia. Methods Sixty-five SD rats were randomly divided into control group, injury group and CNTF group. The optic nerve injury and injury model were induced by adding CNTF. The distal segment of optic nerve injury was obtained on the 7th and 14th day after surgery. The gene expression profile, the expression of the gene and the number of the cells were detected by real-time PCR, immunohistochemistry and HE staining respectively. RESULTS: On the 7th day after operation, 608 differentially expressed genes with up-regulated expression and 417 down-regulated expression were screened in CNTF group, including chromatin configuration, transcriptional regulation, neural stem cells, neural differentiation and development, proliferation and apoptosis, Ion channels, receptors, signal transduction and other related genes. Real-time PCR results validated the reliability of the gene chip results. In the CNTF group, the number of cells in the distal segment of optic nerve injury increased. Nestin, glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), extracellular signal-regulated kinase 1/2 (Erk1 / 2) Segment neurofilament (NF) positive substances increased. Conclusion Extrinsic CNTF promotes the dedifferentiation and axon regeneration of glial cells by regulating the expression of dedifferentiation related genes and proteins in damaged optic nerve.