神经毒素MPP+诱发PC12细胞凋亡的实验研究

采用细胞体外培养、噻唑蓝(MTT)比色法、荧光染色技术、透射电镜、流式细胞技术结合的方法,研究了神经毒素MPP 对PC12细胞增殖和凋亡的影响,为进一步研究药物的神经保护作用提供实验模型.结果表明,MPP 对PC12细胞的诱导凋亡作用呈时间和剂量相关性.0.20mM的MPP 作用于PC12细胞48h后,细胞的存活率降低为60%,细胞核发生凝集和断裂,超微结构发生了一系列的变化,呈现出明显的凋亡特征.流式细胞技术的Annexin v-FTTC和PI双染法显示,活细胞占20.2%,晚期凋亡/坏死细胞占51%,早期凋亡细胞占6.75%.提示,MPP 能够诱导PC12细胞凋亡,是研究神经保护作用药物很好的体外实验模型.

The Experimental Studies of Apoptosis Induced by MPP+ in PC12 Cells

By the techniques cell cultivation outside body, MTT assay, Fluorescence Staining, Transmission Electron Microscopy, and Flow Cytometry, we studied the effects of MPP~+ on proliferation and apoptosis in PC12 cells to provide experimental model for the study of neuroprotective effect. Result indicates that MPP~+ can induce apoptosis in PC12 cells, which showed the dependence on time and concentration. When PC12 cells were treated with 0.20 mM MPP~+ for 48 hours, cell viability was reduced to about 60% as determined by MTT assay, cell nuclear was agglomerated and ruptured, series changes was occurred in ultrastructure. All above results showed distinctive apoptosis characters.Annexin V-FITC apoptosis double-stained kit with Flow Cytometry shows that the visible cells were about 20.2%, late apoptosis/necrosis cells were about 51% and early apoptotic cells were about 6.75% It suggests that MPP~+ can induce apoptosis in PC12 cells, which is a valuable experimental model for the study of neuroprotective effect.