Aβ诱导PC-12细胞凋亡建立阿尔茨海默病细胞模型

目的　用β 淀粉样蛋白 (Aβ2 5- 35)诱导PC 1 2细胞凋亡以建立阿尔茨海默病细胞模型。方法　体外培养大鼠嗜铬细胞瘤PC 1 2细胞 ,以不同浓度Aβ2 5 - 35诱导并于不同时间收获细胞 ,应用MTT法观察细胞活力 ,Fura 2 /AM荧光分析法检测细胞内游离Ca2 +浓度 ,Hoechst332 5 8及碘化丙啶复染分析细胞凋亡。结果　Aβ2 5- 35作用于PC 1 2细胞后 ,细胞活力逐渐下降 ,并呈时间和剂量依赖性 ;同时细胞内Ca2 +浓度显著上升 ;不同浓度Aβ2 5 - 35作用后 ,PC 1 2细胞于不同时间出现凋亡的典型形态学特征 ,该时间比Ca2 +浓度上升约晚 6～ 1 2h。结论　Aβ2 5- 35诱导后PC 1 2细胞活力下降、细胞内Ca2 +浓度上升及细胞凋亡 ,可作为较理想的阿尔茨海默病细胞模型

Setting up Alzheimer disease cell apoptosis model with PC-12 cell induced by Aβ25-35

Objective To set up the Alzheimer disease (AD) cell model using PC 12 cells induced with Aβ 25-35 . Methods The PC 12 cells were cultured in vitro and Aβ 25-35 was added into the medium in different concentrations. Cells were harvested at different periods and then MTT assay was used for observing the cell vitality, Fura 2/AM fluorescence ratio imaging for detecting the intracellular Ca 2+ level, and Hoechst 33258/PI double staining for analyzing the apoptosis. Results After being induced with Aβ 25-35 , the cell vitality was decreased in a time and dose dependent manner and intracellular Ca 2+ concentration increased significantly. The PC 12 cells were found to undergo typical apoptosis induced with different concentrations of Aβ 25-35 at different time points, which was about 6～12 h later than the increase of the intracellular Ca 2+ level. Conclusion Induction of the PC 12 cells with Aβ 25-35 causes decrease of the cell vitality, increase of the intracellular Ca 2+ level and apoptosis of the PC 12 cells. This can be used as a good AD cell model.