贵州医科大学学报

2016, v.41;No.195(12) 1365-1369+1375

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AβOs对大鼠原代海马神经细胞丝裂原活化蛋白激酶信号转导通路的影响
Influence of β-amyloid Peptide Oligomers on Mitogen-activated Protein Kinases in Rat Hippocampal Primary Cultured Neuron

赵亮;官志忠;
ZHAO Liang;GUAN Zhizhong;Department of Microbiology,Guizhou Medical University;Department of Pathology,the Affiliated Hospital of Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Guizhou Medical University,Ministry of Education;Key Laboratory of Medical Molecular Biology,Guizhou Medical University;

摘要(Abstract):

目的:观察β-淀粉样蛋白寡聚体(AβOs)对原代培养大鼠海马神经细胞丝裂原活化蛋白激酶(MAPK)信号转导通路的影响。方法:原代培养大鼠海马神经细胞,用免疫荧光染色鉴定纯度,制备并鉴定AβOs,用不同浓度AβOs(0.25、0.5、1及10μmol/L)处理细胞48 h,采用CCK-8试验检测细胞的存活率,蛋白印迹法(Western blotting)检测细胞中细胞外信号调节蛋白激酶(ERK)、c-Jun氨基末端激酶(JNK)及p38磷酸化及总蛋白表达水平。结果:免疫荧光染色结果显示原代大鼠海马神经细胞的体外培养纯度达85%以上;低于0.5μmol/L AβOs对原代神经细胞无明显毒性作用,但可见phospho-ERK1/2和phospho-JNK蛋白表达升高;1μmol/L及更高浓度的AβOs对原代神经细胞有细胞毒性作用,同时可引起phospho-ERK1/2和phospho-JNK蛋白表达水平均降低,但总ERK1/2及总JNK蛋白表达水平未受明显影响,而phospho-p38及总p38蛋白表达水平与AβOs作用浓度呈负相关。结论:AβOs与MAPK信号通路之间的作用可因AβOs的作用浓度不同而出现差异。
Objective:To investigate the influence of β-Amyloid peptide oligomers(AβOs) on mitogen-activated protein kinases(MARK) signal transduction pathways in primary cultured rat hippocampal neuron.Methods:The primary cultured neurons were isolated,and their purity was checked by immunostaining methods.The AβOs were prepared and identified.The primary cultured neurons were exposed to AβOs at different concentrations(0.25,0.5,1 and 10 μmol/L) for 48 h and the survival rate of cells was detected by CCK-8 test.Western blotting was adopted to detect total protein expression level of extracellular signal regulated protein kinase(ERK),c-Jun N-terminal kinase(JNK) and p38.Results:The results of immunostaining showed that the purity of primary cultured neurons was more than 85%.AβOs at the concentration of less than 0.5 μmol/L had no obvious toxic effect on primary cultured neurons,but prompted the protein expression increase of phospho-ERK1/2 and phosphoJNK.AβOs at the concentration ≥1.0 μmol/L had obvious toxic effect on primary cultured neurons and prompted the protein expression decrease of phospho-ERK1/2 and phospho-JNK,but had no obvious effect protein expression levels of pan-ERK1/2 and pan-JNK.The protein expression levels of phospho-p38 and pan-p38 were negatively correlated to AβOs concentration.Conclusions:The influence of AβOs on MAPK signaling pathway may be different according to different concentration of AβOs.

关键词(KeyWords): 阿尔兹海默病;β-淀粉样蛋白寡聚体;线粒体活化蛋白激酶通路;原代培养神经细胞
Alzheimer's disease;β-amyloid peptide oligomers;mitogen-activated protein kinases;primary cultured neurons

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): 赵亮;官志忠;
ZHAO Liang;GUAN Zhizhong;Department of Microbiology,Guizhou Medical University;Department of Pathology,the Affiliated Hospital of Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Guizhou Medical University,Ministry of Education;Key Laboratory of Medical Molecular Biology,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2016.12.001

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