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目的建立神经生长因子(NGF)对2,5己二酮(HD)诱导的PC12细胞凋亡保护作用的模型,并探讨其作用机制。方法以PC12细胞为神经元的细胞模型,将HD和不同浓度的NGF加入培养的PC12细胞中,四甲基偶氮唑盐检测细胞活性,琼脂糖凝胶电泳检测DNA断裂,流式细胞仪检测细胞凋亡率,Westernblot检测p53蛋白表达,比较各组间的差异。结果随NGF浓度增加,细胞存活率升高,差异有统计学意义(P<0.01);DNA断裂减少;细胞凋亡率减小(P<0.01);p53表达量下降(P<0.05)。结论NGF对PC12细胞可能具有直接的神经营养作用,且可保护PC12细胞免于2,5HD诱导的损伤以及具有抗凋亡作用。
Objective To establish a model of neuroprotective effect of nerve growth factor (NGF) on PC12 cell apoptosis induced by 2,5-hexanedione (HD) and to explore its mechanism. Methods PC12 cells were used as the cell model of neurons. HD and different concentrations of NGF were added to the cultured PC12 cells. The cell viability was detected by MTT assay. The DNA fragmentation was detected by agarose gel electrophoresis. Flow cytometry The apoptosis rate was detected by Western blot and p53 protein expression was compared between groups. Results The cell survival rate increased with the increase of NGF concentration, the difference was statistically significant (P <0.01); DNA fragmentation was decreased; the apoptosis rate was decreased (P <0.01); the expression of p53 was decreased (P <0.05). Conclusion NGF may have a direct neurotrophic effect on PC12 cells and protect PC12 cells against 2,5HD-induced injury as well as anti-apoptotic effects.