关键词: 1-甲基-4-苯基-四氢吡啶离子, 帕金森病, 中脑, 细胞培养, 细胞凋亡

Abstract: Abstract:Objective To investigate the degeneration of dopaminergic neurons induced by MPP⁺ in mouse mesencephalic dissociated culture and establish the cell model for parkinson’[KG-*3]s disease( PD) research in vitro.Methods OF1/SPF mouse embroys of 14 d were used to make mesencephalic dissociated culture,and the culture were divided into control and MPP⁺-treated groups.MPP⁺ with different final concentrations (0.1,1,10 and 15 μmol•L^-1)　 were added into the cultures on the 10th day in vitro,and incubated for 48 h.Then the dopaminergic neurons were identified by TH immunostaining.Hoechst 33342 was used to identify the apoptotic cells.Results The number of dopaminergic neurons was 875±23/well in control group,while there were 612 ±25,586±32,459±16 and 435±19 dopaminergic neurons per well in the 0.1,1,10 and 15 μmol•L^-1 MPP⁺-treated groups respectively,the numbers of dopaminergic neuron decreased significantly in MPP⁺ groups compared with control group（P<0.05）. On the 12th day in vitro,(5.45±0.29 )% cells in control group appeared as typical apoptotic appearance,a higher apoptotic rate was found in MPP⁺ group (26.97%±1.36% )(P<0.05).Conclusion All the concentrations of MPP⁺（0.1,１,10 and 15 μmol•L^-1） could induce the damage and dead of dopaminergic neurons. The degeneration and death are both dose- and time-dependent.Apoptosis may play an important role in the damage of dopaminergc neurons induced by MPP⁺.

Key words: 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine, parkinson’s disease;mesencephalion, cell culture, apoptosis