Investigation on apoptosis of neuronal cells induced by Amyloid beta-Protein

Objective: To construct a PC 12 cell strain with neuronal differentiation, and observe the apoptosis and proliferation activity effects induced these cells by Amyloid beta-Protein (Aβ-43). Methods: 1) PC12 cells in logarithmic growth phase were subcultured for 24 h. After the culture fluid was changed, the cells were treated with Rat-β-NGF and cultured for 9 days. 2) Neuronal differentiation of PC 12 cells in logarithmic growth phase were divided into four groups: control group (0), experimental group (1), experimental group (2) and experimental group (3). The concentrations of Aβ in the four groups were 0 μmol/L, 1.25 μmol/Land 5 μmol/L, respectively. The cells were harvested at 24, 48 and 72 h later and stained with Annexin V-FITC/PI after centrifugation and washing. Then flow cytometry was conducted to examine the apoptosis percentage. 3) NGF-induced PC12 cells were selected and Aβ with different concentrations was added. The final concentrations of Aβ were 0 μmol/L, 1.25 μmol/L, 2.5 μmol/L and 5 μmol/L, respectively. After the cells were incubated in an atmosphere of 5% CO2 at 37 °C in an incubator for 72 h, the OD values were examined. Results: 1) Neuronal differentiated PC12 cell lines were successfully established. 2) Flow cytometric examination indicated that Aβ (1.25, 2.5, and 5.0 μmol/L) could effectively induce apoptosis of neuronal-differented cells at the 24 h, 48 h and 72 h time points. 3) Aβ (0–5.00 μmol/L) had no obvious effect on proliferation or restraining of the neuronal differentiation of the PC 12 cells after a 72 h interacting process. Conclusion: This investigation revealed successful neuronal differentiation of the PC 12 cell strain. The induction of apoptosis of the neurocytes by various concentrations of Aβ was observed and the influence of Aβ on induced proliferation of PC 12 cells by Rat-β-NGF was revealed. This study may provide basis for future research on the molecular cure of AD and interdiction of AD evolution.