一氧化氮对鱼藤酮诱导神经细胞损伤的调节作用

Regulatory role of nitric oxide on rotenone induced neural injury

目的基于SH-SY5Y细胞,建立并优化鱼藤酮诱导的帕金森氏症细胞模型。方法一氧化氮是介导PD病变过程中多巴胺能神经元退变的主要影响因素之一,但采用试剂盒检测鱼藤酮刺激的细胞模型中并未检测到NO的明显改变,使用通用型及神经特异型一氧化氮合酶抑制剂L-NAME及7-NI,均未对鱼藤酮的毒性作用产生任何调节作用。为了研究NO对于鱼藤酮神经损伤模型的调节作用,我们加入了NO供体GSNO与鱼藤酮共处理细胞。结果NO的加入使SH-SY5Y神经细胞转为坏死样死亡形态。GSNO与鱼藤酮共处理与单纯鱼藤酮刺激相比,SHSY5Y细胞中的酪氨酸羟化酶蛋白含量及多巴胺水平降低更显著,细胞线粒体膜电位下调幅度更大,α-突触核蛋白有更显著的增加。结论GSNO的加入能更有效的模拟帕金森氏症中神经元退变的主要特征,是对鱼藤酮诱导模型的有益补充。

Objective Based on SH-SY5Y cells,the cell model induced by rotenone for Parkinson's disease research was established and optimized.Method Nitric oxide is one of the main factors that mediate the degeneration of dopaminergic neurons in the process of PD.However,no obvious change of nitric oxide content was detected in rotenone treated cell model by kit.Both the universal and the neurospecific nitric oxide synthase inhibitors,L-NAME and 7-NI,had no effect on rotenone toxicity.In order to clarify the regulatory effect of nitric oxide on rotenone induced neural injury,we treated SH-SY5Y cells with the nitric oxide donor,GSNO,and rotenone together.Results The dying SH-SY5Y cells took a necrotic shape with the addition of nitric oxide.Compared with rotenone treatment alone,the co-treated cell model showed more significant decrease in tyrosine hydroxylase and dopamine levels,greater decrease in mitochondrial membrane potential and more significant increase inα-synuclein protein content in SH-SY5Y cells.Conclusion The experimental data showed that,the addition of nitric oxide could more effectively simulate the main characteristics of neural degeneration in Parkinson's disease,which would be a beneficial supplement to rotenone induced cell model.