Set up Alzheimer's Disease Cell Apoptosis Model with PC-12 Cell Induced by Aβ25-35
王亚利, 王中卫, 宋潍, YANG LIN
Abstract
王亚利, 王中卫, 宋潍, YANG LIN
Abstract
Objective: To prepare an apoptosis cell model of Alzheimer Disease ( AD ) by PC-12 cells treated with β-amyloid protein (Aβ). Methods: PC-12 cells were incubated with differ-ent concentrations of Aβ25-35 for different duration in vitro. The cell viability was detected by MTT as-say. Morphological features of apoptosis were analyzed with Hoechst 33258/Prorlidium iodide dual staining, The level of intracellular free calcium ([Ca^2+]i) was calculated by Fura-2 / AM fluorescence ratio imaging. Results: ① The viability of PC-12 cells was significantly decreased in prolrrrtion to concentration of Aβ25-35 and duration of exposure to Aβ25-35. ②The apoplotic cells appeared in a time and concentration-dependent manner, and the maximal apoptosis happened at 48 h after execute to 20μmol/ L of Aβ25-35 and 36 h to 30μmol/ L. Cell death reached the peak at 12-24 h later than the apoptotic peak. ③([Ca^2+]i) of PC-12 cells was increased in prolrrrtion to duration of exposure to the same concentration of Aβ25-35. The time of the highest increase rate of [Ca^2+]i was about 12 h earlier than that of apoptosis. Conclusion : An AD cell model using the PC-12 cells induced with Aβ25-35 displays aseries of chanqes related to apoptosis, which may be related to elevation of [Ca^2+]i.
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Objective: To prepare an apoptosis cell model of Alzheimer Disease ( AD ) by PC-12 cells treated with β-amyloid protein (Aβ). Methods: PC-12 cells were incubated with differ-ent concentrations of Aβ25-35 for different duration in vitro. The cell viability was detected by MTT as-say. Morphological features of apoptosis were analyzed with Hoechst 33258/Prorlidium iodide dual staining, The level of intracellular free calcium ([Ca^2+]i) was calculated by Fura-2 / AM fluorescence ratio imaging. Results: ① The viability of PC-12 cells was significantly decreased in prolrrrtion to concentration of Aβ25-35 and duration of exposure to Aβ25-35. ②The apoplotic cells appeared in a time and concentration-dependent manner, and the maximal apoptosis happened at 48 h after execute to 20μmol/ L of Aβ25-35 and 36 h to 30μmol/ L. Cell death reached the peak at 12-24 h later than the apoptotic peak. ③([Ca^2+]i) of PC-12 cells was increased in prolrrrtion to duration of exposure to the same concentration of Aβ25-35. The time of the highest increase rate of [Ca^2+]i was about 12 h earlier than that of apoptosis. Conclusion : An AD cell model using the PC-12 cells induced with Aβ25-35 displays aseries of chanqes related to apoptosis, which may be related to elevation of [Ca^2+]i.
Key concepts: Apoptosis, Viability assay, Programmed cell death, Molecular biology, Cell, Chemistry, Staining, Intracellular