神经元致病蛋白、神经元-胶质细胞网络功能与肌萎缩侧索硬化运动神经元选择性、进行性死亡机制的研究

Characteristic pathologic changes of amyotrophic lateral sclerosis (ALS) are the chosed, progressive motoneron death, however, until to now, its pathogenesis is not clear yet. According to our previous researched results and the newest international researched data, we propose the new presumption of the possible mechanism about the chosed, progressive motoneuron death in ALS be possibly associated with neurodegenerative disease-related proteins (e.g TAR DNA binding protein-43, TDP-43, Fused in sarcoma/translocated in liposarcoma, FUS/TLS, α-Synuclein, α-Syn, Amyloid-β protein, Aβ protein) and function of neuron-glial network.Therefore, we intend to use the ALS transgenic animal model (G93A-SOD1 ALS transgenic mice) of the chosed, progressive motoneuron death, to observe and analyze the expressive, metabolism changes of neurodegenrative disease-related proteins in the different damaged regions of brain and spinal cord, and the distributive changes of them in the internal, external and ultrastructure (Cytosol, mitochondra and nucleus) of the diseased-related neuron and glial cells, and the change of function of neuron-glial network including the change of activation, proliferation, distribution of glial cells and nurotrophic factors release at the different stages of disease by the experimental methodes of fluorescence immunohistochemistry, immunoprecipitation, western blot and image processing etc.. Base on the data of the expressive, metabolism and distributive abnormality of neurodegenerative disease-related proteins and dysfunction of neuron-glial network, we investigate the possible pathogenesis of the chosed, progressive motoneruon death in ALS, to search the new treated methodes of ALS.

肌萎缩侧索硬化(ALS)的特征性病理改变是选择性、进行性运动神经元死亡，至今发病机制尚不清楚。根据我们前期工作基础和最新研究报道推测ALS运动神经元选择性、进行性死亡的机制可能与神经元致病蛋白和神经元-胶质细胞网络功能密切相关。为此，我们选用选择性、进行性运动神经元死亡ALS转基因动物模型(G93A-SOD1 ALS转基因小鼠模型)和荧光免疫组织化学、免疫沉淀、蛋白免疫印迹以及图像处理等技术，观察分析ALS不同发病阶段脑和脊髓受损解剖区神经元致病蛋白的表达、代谢及其在疾病相关的神经元和神经胶质细胞内外、超微结构（线粒体、细胞浆、细胞核）中的分布变化，以及神经胶质细胞的活化、增生、分布和神经营养因子释放等神经元-胶质细胞网络功能的改变。从神经元致病蛋白表达、代谢、分布异常和神经元-胶质细胞网络功能失衡机制，探讨ALS运动神经元选择性、进行性死亡的可能机制，寻找新的ALS治疗靶点。

据我们的前期研究和最新报道：ALS运动神经元选择性、进行性死亡的机制可能与神经元致病蛋白和神经元-胶质细胞网络功能密切相关，我们选用G93A-SOD1 ALS转基因小鼠模型和荧光免疫组织化学以及图像处理等技术，观察分析了ALS不同发病阶段脊髓不同解剖区神经元致病蛋白FUS/TLS与TDP-43的表达、分布和细胞定位，以及神经元-胶质细胞网络功能失衡与运动神经元选择性、进行性死亡的可能关联。我们发现了FUS/TLS蛋白在SOD1 G93A转基因小鼠脊髓星形胶质细胞中表达的增加及异常分布可能引起运动神经元的进行性退变，且星形胶质细胞的损伤可能与运动神经元的死亡密切相关，TDP-43蛋白在SOD1 G93A转基因小鼠脊髓神经元和少突胶质细胞中的过度表达也与运动神经元的死亡呈负相关，可能促进ALS运动神经元的退变，揭示了神经元致病蛋白FUS/TLS、TDP-43的分布表达及定位的异常以及神经元-胶质细胞网络功能失衡在ALS运动神经元选择性、进行性死亡中所起的作用及相关机制，为进一步探究ALS的可能发病机制和治疗方法起到积极的探索作用。