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目的研究TaC lo介导体外培养多巴胺能神经元细胞凋亡及其机制。方法分别用不同浓度的TaC lo(50、100、200、500、1 000μm o l/L)对原代神经元-胶质细胞混合培养和原代神经元培养的细胞进行干预,作用72h,细胞酪氨酸羟化酶(tyrosine hydroxy lase,TH)免疫细胞化学染色后多巴胺神经元计数;分别用浓度为50和200μm o l/L的TaC lo干预混合培养的细胞,作用1、24h后,W estern-b lot法测Caspase-3蛋白表达。结果 TaC lo作用后多巴胺能神经元数量明显下降,50μm o l/L浓度的TaC lo诱发40%的多巴胺能神经元死亡;当TaC lo浓度为200μm o l/L时,接近90%的多巴胺能神经元死亡(P<0.01、P<0.001);TaC lo(50、100、200μm o l/L)诱导的多巴胺能神经元死亡具有明显的剂量依赖性,结果有统计学意义(P<0.001)。W estern-b lot显示,TaC lo作用2个时间点,Caspase-3表达增高,表达均具有药物剂量依赖性。结论 TaC lo可以导致多巴胺能神经元凋亡,这一过程是通过活化的caspase-3蛋白表达等通路完成的。
Aim To investigate TaC lo-mediated apoptosis of dopaminergic neurons in vitro and its mechanism. Methods The primary cultured neurons and glial cells were incubated with different concentrations of TaC lo (50, 100, 200, 500 and 1000μm ol / L) for 72h, Dopamine neurons were counted after immunocytochemical staining with tyrosine hydroxylase (TH); mixed with TaC lo at concentrations of 50 and 200 μmol / L, respectively. After 1 and 24 hours, Western- b lot method to measure Caspase-3 protein expression. Results After TaC lo treatment, the number of dopaminergic neurons decreased significantly. TaC lo concentration of 50 μmol / L induced 40% dopaminergic neuron death. When TaC lo concentration was 200 μmol / L, nearly 90% dopaminergic neurons (P <0.01, P <0.001). The death rate of dopaminergic neurons induced by TaC lo (50,100 and 200μmol / L) was significantly dose-dependent (P <0.001). W estern-b lot showed that TaC lo at two time points, Caspase-3 expression increased, the expression was dose-dependent drug. Conclusion TaC lo can lead to the apoptosis of dopaminergic neurons through the pathway of activated caspase-3 protein expression.