贵州医科大学学报

2016, v.41;No.195(12) 1376-1381

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β-淀粉样肽对大鼠原代海马神经细胞中SIRTs表达的影响
Effect of β-amyloid Peptide on SIRTs Expression in Rat Primary Hippocampal Neurons

董阳婷;谭龙春;官志忠;
DONG Yangting;TAN Longchun;GUAN Zhizhong;Department of Pathology,Affiliated Hospital of Guizhou Medical University;The Key Laboratory of Endemic and Ethnic Diseases in Ministry of Education of P.R.China,Guizhou Medical University;

摘要(Abstract):

目的:探讨β-淀粉样蛋白(Aβ)对原代培养海马神经细胞中沉默信息调节因子(SIRTs)表达的影响。方法:分离出生24 h内SD乳鼠大脑海马区域,培养原代神经细胞,CCK-8试验观察0、0.1、0.2、0.5、1.0、1.5及2.0μmol/L Aβ寡聚体(AβOs)对细胞增殖的影响,选取恰当浓度的AβOs进行后续实验;将实验分为对照组(Control)和Aβ组,Aβ组给AβOs处理24 h后,运用Western blotting方法及Realtime PCR方法检测SIRT1、SIRT3、SIRT4及SIRT5的蛋白及mRNA表达水平。结果:CCK-8结果显示,0.5μmol/L AβOs作用于大鼠原代海马神经细胞24 h时细胞存活率明显低于对照组(P<0.05),且不影响细胞的存活率;0.5μmol/L AβOs处理大鼠原代海马神经细胞24 h后,与Control相比,SIRT1、SIRT3、SIRT4及SIRT5蛋白及mRNA表达水平均不同程度降低(P<0.05或0.01)。结论:AβOs可使海马神经细胞中SIRTs蛋白及mRNA表达水平降低,这可能与AD病人发病后学习记忆能力减退有关。
Objective:To investigate the effect of β-amyloid peptide(Aβ) on expressions of silent information regulators(SIRTs) in rat primary hippocampal neurons.Methods:Rat primary neurons were prepared and cultured from the brain hippocampus of newborn(within 24 h) SD rats.Cell Counting Kit-8(CCK-8) was employed to detect the effect of 0,0.1,0.2,0.5,1.0,1.5 and 2.0 μmol/L Aβ oligomers(AβOs) on proliferation of neurons.The appropriate concentration of AβOs was selected for subsequent experiments.Rat primary hippocampal neurons were divided into control group and Aβ group which was treated with Aβ for 24 h.The protein and mRNA expressions of SIRT1,SIRT3,SIRT4 and SIRT5 were detected by Western blotting and real-time PCR.Results:According to CCK-8 results,0.5 μmol/L AβOs was selected for subsequent experiments.The protein and mRNA expression of SIRT1,SIRT3,SIRT4 and SIRT5 were significantly attenuated when the cells treated with0.5 μmol/L AβOs for 24 h.Conclusions:AβOs can reduce the levels of protein and mRNA of SIRTs in rat primary hippocampal neurons,which might be a mechanism of cognitive deficit of AD.

关键词(KeyWords): 阿尔茨海默病;β-淀粉样肽1-42;海马神经元;沉默信息调节因子
Alzheimer's disease;β-amyloid peptide1-42;hippocampal neurons;silent information regulators

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81260173);; 教育部“长江学者和创新团队发展计划资助”(IRT13058);; 贵州省科技计划[黔科合重大专项字(2014)6008号];; 贵州省创新计划项目[黔教合协同创新中心(2014)06]

作者(Author): 董阳婷;谭龙春;官志忠;
DONG Yangting;TAN Longchun;GUAN Zhizhong;Department of Pathology,Affiliated Hospital of Guizhou Medical University;The Key Laboratory of Endemic and Ethnic Diseases in Ministry of Education of P.R.China,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2016.12.003

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