RNA干扰抑制Nogo-A基因表达对PC12细胞多巴胺释放的影响

INHIBITION OF NOGO-A GENE EXPRESSION BY RNA AND ITS EFFECTS ON DOPAMINE RELEASE IN PC12 CELLS

目的研究Nogo-A基因沉默对PC12细胞多巴胺释放的影响。方法构建靶向Nogo-A的短发夹样RNA(shRNA)真核表达载体,并经脂质体转染PC12细胞,以逆转录聚合酶链反应(RT-PCR)和免疫印迹法检测对Nogo-A mRNA及蛋白的抑制效应。采用高效液相色谱仪(HPLC)检测转染前后多巴胺释放的变化。结果将重组质粒转染到PC12细胞48h后,Nogo-A基因表达明显受抑、多巴胺释放量减少。结论Nogo-A可能参与PC12细胞多巴胺的释放。

Objective To study the effect of Nogo-A gene silence on dopamine release in PC12 cells. Methods The small hairpin RNA（shRNA） eukaryotic expression vector targeting Nogo-A gene was constructed and transfected into cultured PCI2 cells by lipofecamine2000. The inhibiting effect was detected by semi-quantitative reverse transcription（PCR） and Western blotting analysis. The dopamine release was detected by HPLC （high performance liquid chromatography） after transfection. Results The Nogo-A gene expression was inhibited and the dopamine release decresed significantly 48 hours after transfection. Conclusion The reasults suggest that Nogo-A be involved in the mechanism of dopamine realese in PC12 cells.