FK506协同NGF促进缺氧后PC12细胞突起生长的研究及相关机制的探讨

发布时间：2018-03-04 23:08

本文选题：神经再生　切入点：神经生长因子(NGF)　出处：《南华大学》2015年硕士论文　论文类型：学位论文

【摘要】：周围神经损伤在骨科是一种常见的致残性疾病。周围神经损伤后往往伴有神经的缺血缺氧性损伤,特别是挤压伤后。神经再生的过程实际上是受损神经元形成新的轴突或轴突的生长延长,再与靶器官重建突触连接并恢复其正常功能的过程。加快周围神经损伤后轴突的再生,对提高临床周围神经损伤修复后疗效具有重要意义。免疫抑制药物他克莫司(FK506)已被发现在异体手移植的病人上体现出促神经生长的“副”作用,并已表明在实验模型中具有神经保护和神经营养作用,在体内外实验中能增加神经突起的延长和促进神经再生速率。NGF已经被公认在神经损伤后神经再生方面具有神经营养和神经保护作用。本文通过构建神经细胞缺氧模型,研究了他克莫司(FK506)对神经元突起生长的影响,探索了FK506协同神经生长因子(NGF)对神经元突起生长的促进作用,经过一系列的对比实验,筛选出最佳药物配比浓度。进一步定量检测不同浓度FK506在常氧与缺氧状态下,分别对PC12(类神经细胞研究模型)细胞突起生长的影响。并通过蛋白免疫印迹法(Western blotting)检测FK506在常氧与缺氧时对PC12细胞HSP70(热休克蛋白70)、HIF-1ɑ(缺氧诱导因子)表达的影响,探讨FK506保护缺氧性神经元受损的可能性机制。结果表明:在缺氧及常氧状态下,FK506在1-1000 nmol/L均可协同NGF促进PC12细胞突起的生长,其中FK506 100 nmol/L与NGF10ng/ml对缺氧后PC12细胞突起生长的协同作用更明显。100 nmol/L FK506与10ng/ml NGF对相关蛋白表达具有促进作用,100 nmol/L FK506对PC12细胞具有缺氧保护作用,其抗氧化损伤的作用可能与促进HSP70、HIF-1ɑ的表达有关。该研究对促进周围神经再生速率及对神经受损后缺氧性损伤的治疗具有重要的临床意义。
[Abstract]:Peripheral nerve injury is a common disabling disease in orthopedic department. Especially after crush injury, the process of nerve regeneration is actually the process of injured neurons forming new axons or axons, and then reconstructing synaptic connections with target organs and restoring their normal function, and accelerating the regeneration of axons after peripheral nerve injury. The immunosuppressive drug tacrolimus FK506 has been found to play a "secondary" role in promoting nerve growth in patients with hand allograft. It has been shown that there are neuroprotective and neurotrophic effects in the experimental model. NGF has been recognized to have neurotrophic and neuroprotective effects on nerve regeneration after nerve injury in vivo and in vitro. The effects of tacrolimus FK506) on neuronal process growth were studied. The effects of FK506 and NGF on neuronal neurite growth were investigated, and a series of comparative experiments were conducted to investigate the effect of tacrolimus FK506 on neuronal process growth. The optimal drug concentration was screened out. Further quantitative detection of different concentrations of FK506 in normoxic and anoxic state was carried out. The effects of FK506 on the expression of HSP70 (heat shock protein 70) HIF-1 (hypoxia inducible factor) in PC12 cells during normoxic and hypoxia were detected by Western blotting. To explore the possible mechanism of FK506 protecting hypoxic neurons from injury, the results showed that FK506 could promote the growth of PC12 cell processes in combination with NGF at 1-1000 nmol/L under hypoxia and normoxic conditions. The synergistic effect of FK506 100 nmol/L and NGF10ng/ml on the neurite growth of PC12 cells after hypoxia was more obvious. 100 nmol/L FK506 and 10 ng / ml NGF could promote the expression of related proteins. 100 nmol/L FK506 could protect PC12 cells from hypoxia. This study is of great clinical significance in promoting the regeneration rate of peripheral nerve and the treatment of hypoxic injury after nerve injury.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R688

【相似文献】