Aβ寡聚体损伤PC12细胞毒性研究

To study the cytotoxicity of Aβ oligomers on PC12 cells

目的研究Aβ寡聚体对PC12细胞的毒性损伤作用及机制。方法不同比例的Aβ42/Aβ40 Aβ寡聚体作用于PC12细胞构建AD模型,分别应用CCK-8检测细胞活力、ELISA法检测Aβ42蛋白含量,Western blot法检测凋亡相关蛋白Caspase 3表达、自噬相关蛋白Beclin 1及PS1蛋白表达。结果 Aβ42/Aβ40不同比例寡聚体作用于PC12细胞:Aβ42比例越大,Aβ的表达及凋亡越明显,且具有剂量依赖性; PC12细胞活力一定浓度范围呈现增强,超过一定范围后其活力下降; Aβ42/Aβ40对细胞自噬的激活在一定的浓度范围内有效,当浓度过大时,细胞自噬减弱,引起细胞损伤;不同比例Aβ寡聚体损伤PC12细胞,PS1的表达水平无差异。结论 (1) Aβ损伤PC12细胞AD早期细胞模型构建成功;(2) Aβ寡聚体损伤PC12细胞Beclin1蛋白在一定的浓度范围内表达增加,自噬受激活,当Aβ42的浓度达到某一浓度时,Beclin1蛋白的表达减低,自噬被抑制;(3) Aβ寡聚体不改变细胞内PS1的表达水平,其可能通过改变PS1的结构和功能引起Aβ沉积。

Objective To study the toxic damage and mechanism of Aβ oligomers on PC12 cells. Methods PC12 cells were treated with different proportion of Aβ42 with Aβ40 Aβ oligomers to construct AD model. Cell viability was detected by cck-8,protein content of Aβ42 was detected by ELISA,expression of Caspase 3,autophagy related protein Beclin 1 and PS1 were detected by Western blot. Results Aβ42/Aβ40 oligomers with different proportion acted on PC12 cells.The activity of PC12 cells increased in a certain concentration range,and decreased after a certain concentration range. Activation of autophagy by Aβ42/Aβ40 was effective within a certain concentration range,When the concentration was too high,autophagy weakeneds and cell damage was caused. There was no difference in the expression level of PS1 when Aβ oligomers injured PC12 cells at different proportions. Conclusion( 1) The early cell model of Aβ oligomers damaged PC12 cells was successfully constructed.( 2) Beclin1 protein expression in PC12 cells injured by Aβ oligomers increased within a certain concentration range,and autophagy was activated. When Aβ42 concentration reached a certain concentration,Beclin1 protein expression decreased,and autophagy was inhibited.( 3) Aβ oligomers does not change the expression level of PS1 in cells,but may cause deposition of Aβ by changing the structure and function of PS1.