胰岛素样生长因子-1对脊髓源性神经干细胞分化调控的实验研究

发布时间：2018-04-22 01:21

本文选题：脊髓源性神经干细胞 + IGF-1　；参考：《吉林大学》2009年博士论文

【摘要】： 各种原因所致中枢神经损伤后的预后不良,主要是由于神经抑制因子抑制神经再生的作用以及微环境缺乏神经生长因子所导致。神经再生治疗一直是神经损伤研究的热点和前沿,其中对神经干细胞的起源、培养条件以及定向分化调控的研究更是其中的焦点问题。研究发现,成年哺乳动物中枢神经系统内一些非神经发生的部位如脊髓,存在有成体神经干细胞,为脊髓源性神经干细胞(Neural stem cells,NSCs),这为神经再生提供新的易于获得的种子细胞来源,并为神经系统损伤的治疗带来了新的思路和手段。本实验主要研究胰岛素样生长因子-1(Insulin like growth factor-1,IGF-1)对脊髓源性神经干细胞定向分化的调控作用及初步探索其调控机制。IGF-1是一种与组织代谢和细胞分化、增殖有关的细胞因子,对维持神经细胞的生存、生长和损伤后的修复具有极其重要的作用。本实验在体外分离并培养了新生大鼠脊髓源性NSCs;利用脂质体介导法将带有IGF-1基因的质粒载体转入NSCs,经G418筛选,使用RT-PCR、免疫荧光、Western blotting鉴定IGF-1基因的表达,并利用免疫细胞化学法对神经干细胞的分化类型进行鉴定;使用细胞外信号调节激酶(Extracellular signal regulated kinase,ERK)的特异性抑制剂PD98095阻断IGF-1诱导神经干细胞向少突胶质细胞分化,观察神经干细胞的生长状态的变化,使用Western blotting、RT-PCR检测ERK的磷酸化和转录因子c-fos、c-jun、c-myc的表达情况,进而探讨ERK信号途径在IGF诱导神经干细胞分化中的作用和机制。本研究选择IGF-1对脊髓神经干细胞进行干预,观察其作用效果及机制,从而更加深入的探讨脊髓源性神经干细胞在脊髓损伤后的转化方向和影响因素,加深对脊髓损伤后神经再生不良机制的研究,从而为探索治疗脊髓损伤的新方法奠定基础。
[Abstract]:The poor prognosis after central nerve injury is mainly due to the inhibition of nerve regeneration by nerve suppressor and the lack of nerve growth factor in microenvironment. Neural regeneration therapy has been a hot topic and frontier in the research of nerve injury, among which the research on the origin, culture conditions and regulation of directional differentiation of neural stem cells is the focus. The study found that adult neural stem cells were present in some non-neurogenic sites in the central nervous system of adult mammals, such as the spinal cord. It provides a new and easy source of seed cells for neural stem cells and a new method for the treatment of nervous system injury. The aim of this study was to investigate the effects of insulin like growth factor-1IGF-1 on the differentiation of spinal cord derived neural stem cells and to explore its regulatory mechanism. IGF-1 is a cytokine related to tissue metabolism, cell differentiation and proliferation. It plays an extremely important role in maintaining the survival, growth and repair of injured nerve cells. In this study, the spinal cord derived NSCs of newborn rats were isolated and cultured in vitro, the plasmid vector containing IGF-1 gene was transfected into NSCs by liposome-mediated method, and the expression of IGF-1 gene was identified by RT-PCR blotting and G418 screening. The differentiation types of neural stem cells were identified by immunocytochemistry, and the differentiation of neural stem cells into oligodendrocytes induced by IGF-1 was blocked by PD98095, a specific inhibitor of extracellular signal regulated kinase. The growth state of neural stem cells was observed. The phosphorylation of ERK and the expression of c-fos-c-junc-myc were detected by Western blotting RT-PCR, and the role and mechanism of ERK signaling pathway in the differentiation of neural stem cells induced by IGF were discussed. In this study, IGF-1 was chosen to interfere with spinal cord neural stem cells, to observe its effect and mechanism, so as to explore the transformation direction and influencing factors of spinal cord derived neural stem cells after spinal cord injury. To further study the mechanism of nerve regeneration after spinal cord injury, so as to lay a foundation for exploring a new method for treatment of spinal cord injury.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R329

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