溶酶体自噬途径降解α—synuclein蛋白作用研究

目的探讨α—synuclein蛋白细胞内溶酶体途径降解机制。方法用神经生长因子NGF诱导分化PCI2细胞作为研究多巴胺能神经元的细胞载体，应用鱼藤酮处理PCI2细胞建立α-synuclein蛋白细胞模型。使用溶酶体途径降解抑制剂E64处理神经元样分化的PCI2细胞，应用免疫荧光双标方法观察PCI2细胞内硫黄素S、α—synuclein蛋白阳性聚集包涵体形成情况，比较各组的差异。结果用E64处理鱼藤酮预处理过的PCI2细胞后α—synuclein蛋白聚集且较多包涵体形成（15．36±0．85）％，与对照组相比差异有统计学意义（P〈0．05）。结论溶酶体自噬途径可能在α—synuclein蛋白降解、聚集和多巴胺神经元死亡过程中发挥重要作用。

Role of lysosomal autophagy pathway in a-synuclein degradation

Objective To investigate the pathway and mechanism of α-synuclein in lysosomal degradation path way. Methods PC12 cells as dopaminergic neurons, differentiated by nerve growth factor. PCI2 cell5 were treated by rotenone to make α-synuclein model. Then PCI2 cells were treated by lysosomal inhibitors E64. Protein aggregation was detected by the double labeling of thioflavine S and α-synuelein for imunofluoreseenee staining. Results E64 induced α-synuelein protein aggre gate and lead to the formation of eosinophilic intracytoplasmic inclusions, which was immunoreactive to α-synuclein and thioflavine S with （ 15.36 ± 0. 85 ）%（ P〈 0. 05 ）. Conclusion The autophagy pathway plays an important role in the metabolism of a-synuclein, inclusions formation and dopaminergic neuro- nal death.