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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PC12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PC12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PC12细胞,应用免疫荧光双标方法观察PC12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PC12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。
Objective To investigate the intracellular lysosomal pathway degradation mechanism of α-synuclein protein. Methods PC12 cells induced by nerve growth factor NGF were used as the cell carrier to study dopaminergic neurons. The cell model of α-synuclein protein was established by treating rotenone with PC12 cells. Differentiation of PC12 cells into neuron-like differentiated PC12 cells was induced by the lysosomal pathway inhibitor E64. The immunofluorescence double-labeled method was used to observe the formation of positive aggregates of thioflavin S and α-synuclein in PC12 cells. Results Compared with the control group, E64 treatment of rotenone-pretreated PC12 cells resulted in the accumulation of more α-synuclein and more inclusion bodies (15.36 ± 0.85)%, which was significantly different from that of the control group (P <0.05). Conclusion The autophagy pathway of lysosome may play an important role in the degradation and aggregation of α-synuclein protein and the death of dopaminergic neurons.