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目的探讨铜离子对MES23.5多巴胺能细胞的毒性作用及可能机制。方法用MTT法检测铜离子对细胞存活率的影响;用半定量逆转录聚合酶链反应(RT-PCR),Western blotting以及高效液相色谱电化学检测法(HPLC-ECD)检测细胞内酪氨酸羟化酶(tyrosine hydroxlase,TH)mRNA、蛋白的表达以及多巴胺含量的改变;用流式细胞仪检测线粒体跨膜电位的改变。结果100μmol/L和200μmol/L铜离子对细胞存活率没有影响,400μmol/L和800μmol/L铜离子可以造成细胞存活率降低(P<0.01)。200μmol/L铜离子孵育细胞24h,TH mRNA、蛋白表达量以及多巴胺含量较正常对照组降低(P<0.01,P<0.01,P<0.05)。200μmol/L铜离子孵育细胞24h,线粒体跨膜电位较正常对照组明显降低(P<0.01)。结论铜离子对MES23.5多巴胺能细胞具有毒性作用,降低细胞功能的表达,其机制可能与线粒体功能障碍有关。
Objective To investigate the toxic effect of copper ion on MES23.5 dopaminergic cells and its possible mechanism. Methods The effect of copper ion on the cell viability was detected by MTT assay. The expression of tyrosine in the cell was detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and HPLC-ECD, The mRNA and protein expression of tyrosine hydroxlase (TH) and the content of dopamine were detected. The change of mitochondrial transmembrane potential was detected by flow cytometry. Results The cell viability was not affected by 100μmol / L and 200μmol / L copper ions. The cell viability decreased with 400μmol / L and 800μmol / L copper ions (P <0.01). After incubated with 200μmol / L Cu2 + for 24 hours, the expression of TH mRNA, protein and dopamine were lower than those in normal control group (P <0.01, P <0.01, P <0.05). After incubated with 200μmol / L copper ion for 24 hours, the transmembrane potential of mitochondria was significantly lower than that of the normal control group (P <0.01). Conclusions Copper ions have a toxic effect on MES23.5 dopaminergic cells and reduce the expression of cellular functions, which may be related to mitochondrial dysfunction.