乌头碱对胎鼠中脑多巴胺能神经元细胞的神经毒性作用

目的: 观察乌头碱对中脑多巴胺能神经元细胞的毒理作用,以了解和预防乌头碱及其衍生物产生的神经毒性作用。方法: 从孕14 d的小鼠中取出胎鼠中脑,并分离出多巴胺能神经元细胞,于5％CO2,37 ℃和100％湿度条件培养,培养第10天加入不同浓度的乌头碱(0.1,0.5,1,5,10,50 和100 μmol/L)作用48 h,于第12天免疫组化染色。选取神经元细胞数、神经元树突的长度和数量作为观察指标,并进行分析。结果: 乌头碱作用48 h后,不同浓度的乌头碱(0.5～100 μmol/L)对神经细胞生长均有明显的抑制作用,并表现出浓度依赖性神经毒性作用,高浓度乌头碱(10～100 μmol/L)的抑制强度高于低浓度组(0.5～5 μmol/L)。结论: 体外实验证实了乌头碱对中脑多巴胺能神经元有神经毒性作用。

Neurotoxic effect of aconitine based on the midbraindopaminergic neurons cell culture

Objective: To  observe toxicological effects of aconitine on midbrain dopaminergic neurons and to understand and prevent  neurotoxic side effects from aconitine and its derivatives. Methods: Fetal rat brain was taken out from the 14 days pregnant mice and dopaminergic neurons were isolated to culture in 5％ CO2, 37 ℃ and 100％ humidity conditions. Different concentrations of aconitine (0.1, 0.5, 1, 5, 10, 50 and 100 μmol/L) were added on 10th day in vitro and treated for 48 hours. Immunohistochemical staining was done on 12th day and  the number of cells, dendritic length and the number of neuron axons were selected as observed indicators, and to make the analysis. Results: Forty eight hours later, different concentrations of aconitine(0.5-100 μmol/L) had strong inhibition on growth of neuron cells, which showed a significant concentration dependent neurotoxicity. While the high concentrations of aconitine (10～100 μmol/L) had stronger influence on neuron cells than the low concentrations of aconitine (0.5～5 μmol/L). Conclusion: The neurotoxicity effects of aconitine on midbrain dopaminergic neurons in vitro experiments were confirmed for the first time.