脊髓损伤后神经元DNA损害及修复的分子机制研究

脊髓损伤可造成组织内氧化应激升高和炎症因子释放，由此所造成的神经元细胞DNA损害是导致神经元细胞死亡及神经功能紊乱的重要原因。因此揭示脊髓损伤后神经元细胞DNA受损机制、损伤特性及修复机制确认意义重大。神经元细胞DNA受损机制、损伤类型、结构复杂性、分布情况、损伤位点及周边组织中神经元细胞DNA损伤空间及时间相关性，以及各损伤修复通路之间的协同作用，是决定损伤可修复性及其所导致的不良后果的关键因素。本项目拟利用高通量液相色谱、气相色谱质谱联用法对脊髓损伤后脊髓组织内神经元细胞DNA的损伤类型进行确认；利用原子力显微镜确认DNA损伤的结构复杂性、位点分布；利用修复通路敲除型细胞系确认DNA损伤受损及修复机制。通过揭示脊髓损伤所造成的神经元细胞DNA受损机制、损伤类型、结构复杂性、分布情况、时空相关性及修复机制，为脊髓损伤的机制研究及临床康复治疗提供有力的理论支撑及指导。

Neuronal DNA damage induced by oxidative stress increase and inflammatory factors release after spinal cord injury is the critical reason of neurons death and spinal cord tissue physiology variation. Hence, it is of great significance to reveal the properties of spinal-cord-injury induced neuronal DNA damage and mechanisms of DNA damage repair. The type、structure complexity、distribution on DNA strand、spatial and temporal correlation of neuronal DNA damage on injury site and surrounding tissue、mechanisms of DNA damaged and repair are critical factors of DNA damage repairability and destructive consequence. In this study, the types of neuronal DNA damage will be distinguished with high performance liquid chromatography、gas chromatography-mass spectrometry. The structure complexity and distribution on DNA strand of DNA damage will be analyzed by atomic force microscopy. The vital DNA repair pathway、the cooperation between different DNA repair pathway and DNA damaged mechanisms will be intensively researched on correlative gene knockout cell. The verification of type、structure complexity 、distribution of neuronal DNA damage、spatial and temporal correlation, and mechanism of DNA damaged and repair will provide forceful support and theoretical direction to mechanism research and rehabilitation treatment of spinal cord injury.