β-淀粉样蛋白_(25-35)致伤对PC12神经元突触相关蛋白表达的影响  

Effects of amyloid-beta 25-35 on expression of synapse-associated proteins in PC12 neurons

作　　者：张爽[1] 黄昕艳[2] 刘爽[3] 李艳君[3] 赵锦程[3] 

机构地区：[1]佳木斯大学药学院,黑龙江省佳木斯市154007 [2]佳木斯大学附属第二医院,黑龙江省佳木斯市154007 [3]佳木斯大学基础医学院,黑龙江省佳木斯市154007

出　　处：《中国组织工程研究》2016年第2期224-229,共6页Chinese Journal of Tissue Engineering Research

基　　金：黑龙江省教育厅科研项目(1251666);佳木斯大学基础研究重点项目(12Z1201502);佳木斯大学研究生科技创新项目(LZZ2014_030)~~

摘　　要：背景:脑内β-淀粉样蛋白的聚集可诱导神经细胞凋亡,大量神经元及突触的缺失和功能的损害尚无有效的干预手段,提高突触可塑性为治疗早期阿尔茨海默病提供重要方向。目的:筛选最佳的阿尔茨海默病模型,检测β-淀粉样蛋白25-35致伤PC12神经元的突触相关蛋白表达。方法:采用50μg/L神经生长因子诱导PC12细胞分化为神经元样细胞,以不同浓度β-淀粉样蛋白25-35致伤PC12神经元样细胞。应用CCK8法检测细胞生存率。神经颗粒素、神经调节素免疫荧光染色观察模型细胞的形态学变化,Western blot法检测神经颗粒素、CAMKⅡ、PSD-95蛋白表达水平。结果与结论:随着β-淀粉样蛋白25-35浓度增高和作用时间的延长,PC12神经元生存率呈剂量依赖性降低;可见突触长度变短、神经元萎缩、神经元彼此连接疏松;神经颗粒素、CAMKⅡ、PSD-95蛋白表达均下调。结果提示,10μmol/Lβ-淀粉样蛋白25-35、48 h是筛选早期PC12神经元阿尔茨海默病细胞模型的最佳干预浓度和时间。BACKGROUND： An amyloid-beta（Aβ） aggregation in the brain can induce nerve cell apoptosis, loss of synapses and functional damage. However, there is still no effective intervention. Improving the synaptic plasticity provides an important direction for the treatment of early Alzheimer＇s disease. OBJECTIVE： To screen the best model of Alzheimer＇s disease and to explore the expression of synapse-associated proteins in Aβ25-35-injured PC12 neurons. METHODS： PC12 cells were induced by 50 μg/L nerve growth factor to differentiate into neuronal-like cells. Then, these cells were treated with Aβ25-35 at different concentrations. Consequently, cell survival rate was detected using cell counting kit-8; neurogranin and neuregulin immunofluorescence stainings were used to observe morphological changes of model cells; western blot used to detect the expression level of neurogranin, calmodulin kinase Ⅱ, postsynaptic density-95 proteins. RESULTS AND CONCLUSION： Over time, the survival rate of PC12 neurons induced by Aβ25-35 was decreased in a dose-dependent manner. Shortened synaptic length, neuronal atrophy and sparsely interconnected neurons were visible. Expression levels of neurogranin, calmodulin kinase Ⅱ and postsynaptic density-95 proteins were all down-regulated. These findings indicate that to screen the cell model of Alzheimer＇s disease, the optimal concentration and interventional time of Aβ25-35 are 10 μmol/L and 48 hours, respectively.

关 键 词：淀粉样蛋白 突触蛋白类 阿尔茨海默病 组织构建 组织工程 PC12细胞株 Β-淀粉样蛋白 神经元 突触后致密物 突触可塑性 

分 类 号：R318[医药卫生—生物医学工程]