关键词: PC12细胞, α-synuclein, 细胞凋亡, 帕金森病

Abstract: Objective To establish cell model of Parkinson’〖KG-3〗s disease (PD) and to approach the toxic effect of rotenone on dopaminergic neurons and its mechanism . Methods PC12 cells were differentiated into dopaminergic neurons and treated with various concentrations of rotenone. The morphological changes of PC12 cells were observed after treated with rotenone（0,10,25,50,75, and 100 nmol•L^-1）for 24,48 and 72 h , the cell viability was measured by MTT assay . Immunohistochemistry and immunofluorescence were used to observe the accumulation of α-synuclein in cytoplasm. AO/EB double staining was also adopted to test apoptosis. Results After differentiation PC12 cells shaped irregularly with big and long ecptomas and multiple cell conjunctions. After the treatment of rotenone cell ecptomas vanished gradually and cell bodies became smaller and smoother. The cell viability began to decline significantly at a concentration of 50 nmol•L^-1 for 24 h (P<0.05) ，and as concentration increased, the cell viability declined in a dose-dependent manner (P<0.01).As time prolonged, the cell viability behaved the same way. There was a significant difference between each time groups (P<0.01) .Immunohistochemistry demonstrated a brown round α-synuclein immunoactive mass in cytoplasm; cell immunofluorescence showed a strong greenα-synuclein immunoactive aggregation in cytoplasm. Viable apoptotic cells，non-viable apoptotic cells and necrosis cells were observed gradually as cell concentration increased and time prolonged. Conclusion Rotenone should be toxic to dopaminergic neurons，causing inclusion of α-synuclein aggregation and inducing apoptosis. The malmetabolism of α-synuclein and apoptosis induced by rotenone might play an important role in the pathogenisis of Parkinson’〖KG-3〗s disease .

Key words: PC12 , α-synuclein , apoptosis , Parkinson’s disease