戊四氮及NF-κB圈套结构对神经元样细胞GAP-43表达的影响

Effects of Pentylenetetrazol and Nuclear Factor kappa B Decoy Oligodeoxynucleotides on the Expression of Growth-associated Protein-43 in Neuron-like PC12 Cells

目的:观测戊四氮(PTZ)及NF-κB圈套寡核苷酸结构对神经元样PC12细胞生长相关蛋白-43(GAP-43)表达的影响。方法:应用免疫印迹检测PTZ干预前后神经元样PC12细胞GAP-43的表达情况,进而运用基因转染技术将NF-κB圈套寡核苷酸转入神经元样PC12细胞中,应用激光共聚焦成像技术(CLSM)检测转染前后NF-κB的活性变化,免疫印迹观察转染前后GAP-43的变化情况。结果:①PTZ干预后神经元样PC12细胞GAP-43的表达显著增高;②CLSM检测显示转染NF-κB圈套寡核苷酸后神经元样PC12细胞中NF-κB活性明显下降,但GAP-43表达水平未降低。结论:PTZ可促进GAP-43的表达,NF-κB对GAP-43的表达不具有调控作用。

Objective.. To investigate the effects of pentylenetetrazol （PTZ） and nuclear factor kappa B decoy oligodeoxynucleotides （NF-κB ODNs） on the expression of growth associated protein-43 （GAP-43）in neuron-like PC12 cell and to explore the epileptogenesis. Methods：The expression level of GAP-43 was studied by Western blot in neuron-like PC12 cells with and without PTZ treatment. NF-κB decoy ODNs were transfected into neuron-like PC12 ceils with Lipofectamine-2000, and confocal laser scanning microscope （CLSM） was performed to investigate the activation of NF-κB. Results： （1） Western blot showed that the level of GAP-43 was significantly increased in neuron-like PC12 cells with PTZ treatment when compared to the control cells. （2） CLSM showed that the decoy ONDs decreased the activation of NF-κB in neuron-like PC12 cell, while Western blot showed that the decoy ONDs did not decrease the level of GAP-43. Conclusion： PTZ can enhance the expression of GAP-43 and active NF-κB in PC12 cells. NF-κB could not modulate GAP 43 expression.