锰致PC12细胞PARK2表达对多巴胺分泌的修饰作用
Modification of Dopamine Secretion in PC12 Cells by Manganese Induced PARK2 Gene Expression
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摘要:
目的 研究锰在不同剂量和不同时间下对鼠嗜铬神经瘤细胞(PC12)中多巴胺含量和PARK2 表达的影响。 方法 鼠PC12 细胞分别以(1)0、100、300、500 μmol/L 浓度的氯化锰(MnCl2)染毒24 h; (2)500 μmol/L MnCl2染毒6、24、48 h。采用反相高效液相-荧光法测定细胞内多巴胺含量, 实时荧光定量聚合酶链反应检测PARK2 mRNA水平。 结果 随着染毒浓度的增高, 多巴胺含量及PARK2 mRNA表达呈下降趋势。与对照组比较, 300 μmol/L 或以上浓度MnCl2 染毒所致多巴胺含量降低和PARK2 mRNA表达降低(P < 0.01)。500 μmol/L MnCl2 分别染毒6、24、48 h, 多巴胺含量明显降低(P < 0.01), PARK2 mRNA表达下调(P < 0.05)。 结论 锰可致PC12 细胞多巴胺含量减少, 可能与PARK2的表达下调有关。 Abstract:Objective To study the effect of manganese on the content of dopamine and the expression of PARK2 in pheochromocytoma (PC12) cells at different doses and different time lengths. Methods Rat PC12 cells were incubated for 24 h with 0, 100, 300, 500 μmol/L of MnCl2, and incubated with 500 μmol/L MnCl2 for 6, 24, and 48 hours. The content of dopamine in PC12 cells were determined by reverse phase high performance liquid chromatography combined with fluorometric detection. The expression of PARK2 mRNA was examined by real time quantitative polymerase chain reaction. Results The dopamine content and PARK2 mRNA expression level tended to decline with the increasing MnCl2 concentrations. Compared with the control group, after 24 h of incubation, the 300 and 500 μmol/L MnCl2 significantly decreased the dopamine content and PARK2 expression (P < 0.01). When the PC12 cells were exposed to 500 μmol/L Mncl2 for 6, 24, and 48 h respectively, the content of dopamine and the expression of PARK2 mRNA were also significantly lowered (P < 0.01, P < 0.05 respectively). Conclusion Manganese could reduce dopamine content in PC12 cells, and the effect may be associated with down-regulated PARK2 expressions.
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