帕金森病多巴胺能神经元轴突变性新的体外实验细胞模型的建立  

Establishment of an in vitro cell model of axonal degeneration of dopaminergic neurons in Parkinson＇s disease

作　　者：赵海康[1] 李立宏[2] 潘力[3] 于军[1] 方伟[2] 夏毅[2] 汪鑫[2] 高国栋[2] 

机构地区：[1]西安医学院第二附属医院神经外科,710038 [2]第四军医大学唐都医院神经外科 [3]广州军区武汉总医院神经外科

出　　处：《中华实验外科杂志》2015年第9期2286-2291,共6页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81270329）;陕西省社发攻关项目（2012K-02-01）;陕西省教育厅科研项目（12JK0706）;陕西省卫生厅科研项目（2012D4）

摘　　要：目的 建立帕金森病（PD）多巴胺能神经元轴突变性新的体外实验细胞模型.方法 检测不同浓度1-甲基-4-苯基吡啶离子（MPP＋）致原代培养的小鼠中脑腹侧多巴胺神经元损伤的能力.采用免疫组织化学的方法检测轴突的形态学变化、存活神经元的数目、轴突的长度和数目,以及凋亡多巴胺神经元的数目.噻唑蓝（MTT）比色检测细胞活力在MPP＋处理2～24h的不同时间点进行.结果 10.0μmol/L的MPP＋作用24 h导致多巴胺神经元显著的轴突变性.神经元的数目、轴突的长度和数目均分别减少50.39％、59.80％、35.70％.结论 10.0 μmol/L的MPP＋作用24 h造成的体外实验模型可以有效地反映帕金森病的轴突变性.Objective To establish an in vitro model of the axonal degeneration of dopaminergic neurons in parkinson＇ s disease （PD）.Methods A variety of concentrations of 1-methyl-4-phenylpyridinium （MPP ＋） were tested for their ability to induce injury of primary cultured dopaminergic neurons from the mouse ventral mesencephalon.Morphological changes to axons,the number of remaining neurons,the length and number of axons,and the number of apoptotic dopaminergic neurons were observed by immunohistochemistry,methyl thiazol tetrazolium （MTT） assays was performed to determine the cell viability at different time points from 2 h to 24 h after addition of MPP ＋.Results Exposure of dopaminergic neurons to 10.0 μmol/L MPP ＋ for 24 h caused significant axonal degeneration.The number of neurons,axon length and the number of axons were respectively decreased by 50.39％,59.80％,and 35.70％.Conclusion An in vitro model in which cells are treated with 10.0 μmol/L MPP＋ for 24 h can be used as an effective model of the axonal degeneration of dopaminergic neurons in PD.

关 键 词：多巴胺神经元 1-甲基-4-苯基吡啶离子 神经元轴突变性 帕金森病 

分 类 号：R742.5[医药卫生—神经病学与精神病学]