戊四氮及NF-κB圈套结构对神经元样细胞突触素表达的影响

Effects of Pentylenetetrazol and Nuclear Factor kappa B Decoy Oligodeoxynucleotides on the Expression of Synaptophysin in Neuron-like PC12 Cells

目的通过观测戊四氮(pentylenetetrazol,PTZ)及NF-κB(nuclear factor kappa B decoy)圈套寡核苷酸结构对神经元样PC12细胞突触素(synaptophysin,P38)表达的影响,进一步探讨癫痫的发病机制。方法应用免疫印迹技术检测PTZ干预前后神经元样PC12细胞突触素的表达情况,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,激光共聚焦成像技术(confocal laser scanning microscope,CLSM)检测转染前后NF-κB活性的变化,免疫印迹观察转染前后突触素的变化情况。结果①PTZ干预后神经元样PC12细胞突触素的表达显著增高;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,但突触素蛋白表达水平不见降低。结论PTZ促进了突触素的表达,NF-κB对突触素的表达不具有调控作用,可能另有其他途径。

Objective To investigate the effects of pentylenetetrazol （PTZ） and nuclear factor kappa B decoy oligodeoxynucleotides （NF-κB ODNs） on the expression of synaptophysin in neuron-like PC12 cells and explore the epileptogenesis. Methods The expression level of synaptophysin was observed by Western blot in neuron-like PC12 cells treated with or without PTZ. NF-κB decoy ODNs were transfected into neuron-like PC12 cells with Lipofectamine-2000, and by using confocal laser scanning microscope （CLSM） the activation of NF-κB was investigated, and the expression level of synaptophysin was detected by Western blot before and after transfection. Results ① Western blot revealed that the level of synaptophysin was increased significantly in neuron-like PC12 cells treated with PTZ as compared with that in 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 synaptophysin. Conclusion PTZ could enhance the expression level of synaptophysin and activate NF-κB in PC12 cells. NF-κB could not modulate the expression level of synaptophysin.