贵州医科大学学报

2016, v.41;No.191(08) 878-881

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一种改进的Aβ_(1-42)寡聚体的制备与鉴定方法
An Improved Method for Preparation and Identification of Aβ_(1-42) Oligomer

禹文峰;孔欣;官志忠;
YU Wenfeng;KONG Xin;GUAN Zhizhong;the Key Lab of Medical Molecular Biology,Guizhou Medical University;Department of Pathology,Guizhou Medical University;

摘要(Abstract):

目的:改进Aβ_(1-42)寡聚体的制备与鉴定方式。方法:以化学合成多肽Aβ_(1-42)为原料在体外制备Aβ_(1-42)寡聚体,用Western blotting进行鉴定;加入不同浓度Aβ_(1-42)寡聚体后,用CCK-8法测定体外原代培养大鼠大脑皮质星形胶质细胞(AS)的存活率。结果:化学合成的Aβ_(1-42)寡聚体分子量为4~188 k D,以64 k D最多,亦有少量4 k D单体存在;CCK-8法显示Aβ_(1-42)寡聚体对AS有明显的神经毒性作用,并呈现剂量-效应关系。结论:成功建立了一种改进的制备和鉴定Aβ_(1-42)寡聚体的方法,为深入研究阿尔茨海默病提供了研究手段。
Objective: To improve method for preparation and identification of Aβ_(1-42) oligomer.Methods: Aβ_(1-42) oligomer was prepared by using chemical synthetic Aβ_(1-42) polypeptide in vitro. The prepared peptide was identified by the assay of Western blotting. In order to confirm the neurotoxicity of the peptide,primary cultured astrocytes from the cortex of rats were treated with different concentrations of Aβ_(1-42) oligomer and the survival rates of the cells were detected via Cell Counting Kit-8( CCK-8) assay. Results: The results indicated that chemical synthetic Aβ_(1-42) oligomer weighed from 4 to188 kd,in which the points with 64 kd were the most. In addition,a few points with 4 kd,a monomer,was also found. Furthermore,the neurotoxicity of Aβ_(1-42) oligomer on astroytes was significantly detected with a manner of dose-dependence. Conclusions: An improved method for preparation and identification of Aβ_(1-42) oligomer has been successfully established,which is important in the research work of Alzheimer's disease.

关键词(KeyWords): 淀粉样蛋白;阿尔茨海默病;寡聚体;星形胶质细胞;免疫印迹法
amyloid;Alzheimer disease;oligomer;astrocytes;immunoblotting

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81360199);国家自然科学基金(81260173);; 教育部科学技术研究项目(213032A);; 贵州省科技厅国际合作项目[黔科合外G字(2011)7014号];; 贵州省科技计划[黔科合重大专项字(2014)6008号]

作者(Author): 禹文峰;孔欣;官志忠;
YU Wenfeng;KONG Xin;GUAN Zhizhong;the Key Lab of Medical Molecular Biology,Guizhou Medical University;Department of Pathology,Guizhou Medical University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2016.08.003

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