一种稳定阿尔茨海默病细胞模型的建立方法  

A method for establishing a stable cellular model of Alzheimer's disease

作　　者：曹方圆[1] 罗德欣[2] 黄玉雕[2] 

机构地区：[1]哈尔滨医科大学基础医学院形态实验中心,黑龙江大庆163319 [2]哈尔滨医科大学附属第五医院神经内科,黑龙江大庆163319

出　　处：《中风与神经疾病杂志》2017年第3期221-225,共5页Journal of Apoplexy and Nervous Diseases

摘　　要：目的旨在应用不同浓度Aβ_(1-42)诱导新生SD大鼠海马神经元,建立稳定阿尔茨海默病细胞模型。方法分离并培养新生24 h内SD大鼠原代海马神经细胞;分别加入不同浓度Aβ_(1-42)诱导,诱导后采用MTT法观察细胞活力,选取理想Aβ_(1-42)诱导的海马神经细胞作为阿尔茨海默病细胞模型;通过免疫荧光技术(IF)鉴定神经元特异性烯醇化酶(NSE)在海马神经元细胞的表达。结果 Aβ_(1-42)诱导后的海马神经细胞存活率下降,可导致海马神经元细胞胞体变形、收缩和细胞膜完整性破坏,胞间网络结构消失或断裂,细胞外基质乱不清,神经元大量凋亡;Aβ_(1-42)诱导后的海马神经细胞存活率下降与Aβ_(1-42)浓度成线性关系;加入不同浓度Aβ_(1-42)(0μmol/L、10μmol/L、20μmol/L、30μmol/L、40μmol/L和50μmol/L)后实验各组细胞存活率分别为(100.00±0.00)%、(98.55±1.42)%、(90.37±3.25)%,(66.34±2.76)%、(53.23±3.41)%和(41.55±2.37)%,其中20μmol/L组与0μmol/L组相比P<0.05,30μmol/L、40μmol/L和50μmol/L组与对照组相比P<0.01。结论 Aβ_(1-42)可导致海马神经元大量凋亡;Aβ_(1-42)浓度为20μmol/L诱导的SD大鼠海马神经元可作为理想的阿尔茨海默病细胞模型。Objective Aimt to establish a stable model of Alzheimer＇s disease by using different concentrations of Aβ1-42to induce hippocampal neurons of the primary culture for hippocampal neurons of SD newborn rats. Methods Extract and culture the primary hippocampal neurons from newborn SD rats within 24 h SD rats; Add to different concentrations of Aβ1-42to induce the hippocampal neurons and observe cell viability by method of MTT,then choose the ideal hippocampal neurons as Alzheimer＇s disease cell model; Use the Immunofluorescence（ IF） to identify the expression of neuron-specific enolase（ NSE） in hippocampal neurons. Results The protein of Aβ1-42could lead to body deformation,shrinkage and cell membrane integrity damage of hippocampal neuron cell,network structure disappears or fracture between cells,clutter unclear of extracellular matrix and apoptosis of a large number of neurons; The survival rate of hippocampal neurons significantly reduced after induced by the protein of Aβ1-42and it had a liner relationship between the concentration of Aβ1-42the cells of SD rats could as a kind of ideal model of Alzheimer＇s disease cells. The survival rates of experimental groups were（ 98. 55 ± 1. 42） %,（ 90. 37 ± 3. 25） %,（ 66. 34 ± 2. 76） %,（ 53. 23 ± 3. 41） % and（ 41. 55 ± 2. 37） % by adding different concentrations of Aβ1-42 Conclusion The protein of Aβ1-42could lead to apoptosis of a large number of neurons;The hippocampal neurons of SD rats can as a kind of ideal model of Alzheimer＇s disease cells,who the induce concentration of Aβ1-42is 20 μmol/L.

关 键 词：阿尔茨海默病 AΒ1-42 海马神经元 模型 

分 类 号：R749.16[医药卫生—神经病学与精神病学]