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目的 探讨 L- dopa在帕金森氏病 (PD)治疗中的神经毒性作用及其毒性机制。方法 以 PC12细胞为多巴胺神经元的细胞模型 ,利用 DNA凝胶电泳及图像分析仪、TU NEL 法、流式细胞术及免疫荧光技术检测不同浓度的 L- dopa对 PC12细胞的凋亡诱导作用及凋亡相关基因 Bcl- 2、Bax表达的改变。结果 2 0 μmol/ L、5 0 μmol/L、10 0 μm ol/ L、15 0 μmol/ L 处理组凋亡率分别为 2 .5 %、12 .4%、2 4.4%、37.2 % ;5 0 μm ol/ L～ 15 0 μmol/ L 处理组电泳后出现梯状带 ,DNA片断化比例与流式细胞术结果基本一致 ;TUNEL 法检测到的凋亡率高于前两者 ;L- dopa处理后 ,Bcl- 2的表达量减少 ,与凋亡率呈显著负相关 ;Bax的表达量增加 ,与凋亡率呈显著正相关。结论 L- dopa可能是通过凋亡这条途径损害多巴胺神经元的 ,其机制可能是通过改变 Bcl- 2 / Bax的比值来介导细胞凋亡。
Objective To investigate the neurotoxicity of L-dopa in Parkinson’s disease (PD) and its toxicity mechanism. Methods PC12 cells were used as the cell model of dopamine neurons. The apoptosis induction of PC12 cells with different concentration of L-dopa was detected by DNA gel electrophoresis and image analyzer, TU NEL method, flow cytometry and immunofluorescence staining. Changes of apoptosis related genes Bcl-2, Bax expression. Results The apoptotic rate was 20.5%, 12.4%, 24.4% and 37.2% in the groups of 0 μmol / L, 50 μmol / L, 100 μmol / L and 150 μmol / Ladder bands appeared after electrophoresis in 0 μmol / L ~ 150 μmol / L groups, and the proportion of DNA fragmentation was basically the same as that of flow cytometry. The apoptosis rate detected by TUNEL method was higher than that of the former two groups. L-dopa After treatment, the expression of Bcl-2 decreased, and the apoptosis rate was significantly negatively correlated; Bax expression increased, and the apoptosis rate was significantly positively correlated. Conclusion L-dopa may damage dopaminergic neurons through this pathway of apoptosis, and its mechanism may be through the change of Bcl-2 / Bax ratio to mediate apoptosis.