锰对大鼠纹状体神经细胞PARK2表达的影响

Effects of manganese on PARK2 expression in rat striatal neurons

目的研究锰在RNA干扰条件下对大鼠纹状体神经细胞PARK2基因的影响。方法使用不同浓度(0、100、300和500μmol/L)的锰处理感染PARK2 shRNA的原代大鼠纹状体神经元细胞24h,采用实时荧光定量和蛋白印迹法检测基因mRNA水平和蛋白质表达的变化。结果在单纯感染慢病毒的纹状体神经细胞处理组中,不同锰处理浓度组与对照组比较300和500μmol/L锰浓度处理24h后,PARK2 mRNA及蛋白表达显著下降(P<0.01)。在感染慢病毒shRNA的纹状体神经细胞处理组中:不同锰处理浓度与对照组比较,PARK2 mRNA(P<0.01)及蛋白表达均显著下降(P<0.05)。结论转染慢病毒载体PARK2 shRNA的纹状体神经元细胞染锰24h后进一步降低了PARK2 mRNA和蛋白的表达,该作用过程可能是锰致神经系统危害的作用机制之一。

Objective To explore the effects of manganese（Mn） on PARK2 expression in rat striatal neurons with/without the RNA interference.Methods After treatment with a serials of manganese concentration（0,100,300,500 μmol/1） for 24 h,the PARK2 mRNA and protein（Parkin） levels,in the rat striatal neurons（transfected with lentivirus or with lentivirus PARK2 shRNA）,were measured by real-time PCR and western blot,separately.Results Compared with the control group,the levels of PARK2 mRNA and Parkin decreased significantly（P〈0.01） in the striatal neurons transfected with lentivirus after exposure to 300 and 500 μmol/1 of Mn for 24 h,while the decreases were more significant（P〈0.05,P〈0.01） in striatal neurons（transfected with lentiviral PARK2 shRNA） after exposure to 100,300 and 500 μmol/1 of Mn for 24 h.Conclusion The striatal neurons transfected with lentiviral PARK2 shRNA expressed lower levels of PARK2 mRNA and Parkin protein after exposure to Mn for 24 h,indicating that Mn suppresses PARK2 expression as a mechanism of its toxicity to neurons.