睫状神经营养因子调节受损视神经胶质细胞去分化相关基因的表达  

Ciliary neurotrophic factor regulates genes expression involved in dedifferentiation of glial cells in injured optic nerves of rats

作　　者：郭金萍[1] 许家军[1] 刘芳[1] 袁建明[1] 

机构地区：[1]第二军医大学解剖学教研室,上海200433

出　　处：《解剖学报》2011年第3期289-295,共7页Acta Anatomica Sinica

基　　金：国家重点基础研究发展计划资助项目(2011CB707502);国家自然科学基金资助项目(30271392)

摘　　要：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 mechanism underlying ciliary neurotrophic factor （CNTF） involved in dedifferentiation of glial cells in injured optic nerves. Methods Sixty-five adult male SD rats were randomly divided into 3 groups： normal control group, optic nerve injured group and CNTF treat group. Optic nerves were harvested at the 7th or the 14th day after surgery and detected by gent chip, real-time PCR, HE staining and immunohistochemistry. Results Comparison with those of injury group, there were 608 genes expression up-regulated and 417 ones down-regulated in the CNTF treated group at the 7th day after operation, including genes related to chromatin configuration, transcription regulation, neural stem cells, neural differentiation and development, proliferation, apoptosis, ion channel, receptor, signal transduction and so on. Result of real-time PCR for selected genes was consistent with that of gene chip, supporting the validity of gene chip data. We also found that cells revealed by HE staining and immunoreactivity of Nestin, myelin basic protein （MBP） , Erkl/2, and glial fibrillary acidic protein （GFAP） were increased in the distal optic nerves, but immunoreactivity of neurofilament （NF） was increased in the proximal optic nerves in the CNTF treated group compared with the optic nerve injured group. Conclusion CNTF induces dedifferentiation of glia cells in the injured optic nerves and promotes regeneration of injured optic nerves through regulating the expression of related genes and proteins.

关 键 词：视神经 损伤 睫状神经营养因子 神经胶质细胞 去分化 基因芯片 免疫组织化学 大鼠 

分 类 号：R774.6[医药卫生—眼科]