过量Zn染及缺血氧处理对神经细胞离子通道电流的影响-动物学专业论文.docxVIP

摘要摘要 摘要 摘要 Zn2+稳态维持对脑内神经元行使其生理功能起着至关重要的作用，过量的 Zn2+会对神经元造成损伤甚至促发神经元级联性死亡。另外，缺血缺氧状态极 易导致神经元细胞崩解，神经结构与功能损伤。本实验采用PCI2细胞及新生 大鼠海马神经元两种细胞，通过电生理学实验手段，研究过量zn染及缺血缺氧 对神经细胞电压依赖性离子通道电生理特性的影响，以期在离体状态下能为在 胞外过量Zn2+、缺血缺氧状态引发的神经元损伤的相关机制提供一些依据，可 望为一些神经性疾病的发病机制、防治及神经保护措施的提出提供些许思路。 主要实验内容和结果如下： 1．使用急性分离手段分离大鼠海马神经元细胞，获得的细胞胞体圆润饱满， 细胞膜完整、折光性好，符合电生理实验要求。 2．使用膜片钳技术，记录到PCI2细胞和急性分离的海马神经元4-AP敏感的 厶和TEA敏感的k，以及海马神经元TTX敏感的氐。。 3．使用连二亚硫酸钠联合无糖培养基处理(即OGD模型)，建立了PCI2细 胞和大鼠海马神经元缺血缺氧损伤模型。 4．OGD处理PCI2细胞和大鼠海马神经元，使细胞存活率降低，且呈时间、 浓度依赖性，产生神经毒害作用。 5．过量Zn染或OGD单一处理均降低PCI2细胞和大鼠海马神经元厶和k的 幅值，使厶和k的I．V曲线下移，k激活曲线左移；而过量Zn染与OGD联合 处理比过量Zn或OGD单一处理使上述变化趋势更为明显。 根据上述结果得出以下结论：胞外过量zn染和／或缺血缺氧状态可造成神 经元损伤，且缺血缺氧可加重过量Zn2+对神经元的损伤作用，其机制可能与其 对厶和k的影响而使神经元过度兴奋有关。 关键词：锌，氧糖剥夺，电压门控离子通道，膜片钳，神经元损伤 Abstract Abstract Abstract The homeostasis of neuron Zn+plays a vital role in the physiological function of brain．The excessive extracellular zinc would cause neural damage and trigger serial actions of neuron death．In addition，ischemia and hypoxia can cause cell collapse，lead to damage of the structure and function in nerve system．In this study, PC 1 2 cell and cultured hippocampus cell are used to study the effect of excessive zinc and ischemia—hypoxia Oil voltage—gated ion channels of neural cells，in order to clarify the involved mechanisms of the neural damage creased by excessive zinc， ischemia-hypoxia and other neurological diseases． The main contents and results in present study included： 1．Hippocampal neuron fitting for electrophysi0109ical experiments was isolated successfully from new—bom rat by procedure of acute separation． 2．4-AP sensitive厶and TEA sensitive k of PC 1 2 and hippocampal neuron were recorded by patch clamp technique；TTX sensitive／sa of hippocampal neurons was also recorded． 3．The hypoxic ischemic model of PC 1 2 and hippocampal neuron was established by oxygen—glucosed deprivation． 4．OGD treatment decreased survival ratio of PC 1 2 cells and hippocampal neurons with time and concentration dependent，showed serious neurotoxici