氧化应激及黑色素对神经细胞铁蛋白mRNA表达的影响

The effects of oxidative stress on ferritin mRNA gene expression

目的 探讨神经黑色素（NM）对神经元和神经胶质细胞在氧化应激状态下铁蛋白mRNA表达的影响. 方法 采用人类神经母细胞瘤细胞株（SK-N-SH）和人类胶质母细胞瘤细胞株（U373）作为神经元和神经胶质细胞模型,以Fenton试剂（FR）诱导氧化应激后分别加入NM或多巴胺黑色素（DAM）,同时将铁螯合剂去铁胺（DFO）处理过的细胞作为对照,Q-PCR检测各组铁蛋白mRNA的表达. 结果 FR、FR＋DAM处理后SK-N-SH细胞铁蛋白mRNA的表达高于空白对照组,FR＋DAM组铁蛋白mRNA表达高于FR＋NM组,差异均有统计学意义（P＜0.05）；与DFO未处理组比较,FR、NM、FR＋NM、DAM组中相对应的DFO处理组铁蛋白mRNA的表达均明显降低,差异有统计学意义（P＜0.05）; FR、NM、DAM、FR＋NM和FR＋DAM处理后U373细胞铁蛋白mRNA的表达无明显变化,DFO处理组与相应的DFO未处理组比较铁蛋白mRNA的表达也无明显变化,差异无统计学意义（P＞0.05）. 结论 铁蛋白对细胞有保护作用,DAM不能模拟NM制作PD模型.

Objective To explore the effects of human neuromelanin （NM） and doparmine melanin （DAM） on ferritin mRNA expression under oxidative stress in dopaminergic neuronal cells and glial cells. Methods SK-N-SH and U373 were chosen as models of human neuronal and glial cells.Fenton reagent （FR） was used as a direct inducer of oxidative stress,and human neuromelanin （NM） or dopamine melanin （DAM） were added to the cultures at the same time. Iron chelator desferrioxamine （DFO） was added in some experiments parallel.The changes of ferfitin mRNA expression were detected by real-time quantitative PCR. Results Ferritin mRNA in SK-N-SH cells were significantly up-regulated with FR or FR＋DAM treatment than untreated cells,and FR＋DAM induced more Ferritin mRNA expression than FR＋hNM did.These differences are significant （P〈0.05）.With DFO,ferritin mRNA expressions in FR, NM,FR＋NM, DAM treated SK-N-SH cells were decreased significantly comparing to DFO free cultures（P〈0.05）.While in U373 cells,the expressions of Ferritin mRNA had no significant differences with or without FR,NM, DAM, FR＋NM or FR＋DAM treatment （P〈0.05）.Furthermore,Ferritin mRNA expressions showed no significant differences in U373 cells with or without DFO treatment（P〈0.05）. Conclusions Ferritin has a protective effect on cells,DAM could not used to produce PD model.