一种稳定阿尔茨海默病细胞模型的建立方法

发布时间：2019-04-21 12:32

【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P0.05,30μmol/L、40μmol/L和50μmol/L组与对照组相比P0.01。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。
[Abstract]:Aim to establish a stable Alzheimer's disease (AD) cell model by inducing hippocampal neurons in neonatal SD rats with different concentrations of A 尾 _ (1 尾 _ (42). Methods Primary hippocampal neurons of SD rats were isolated and cultured within 24 hours after birth. After induction with different concentrations of A 尾 _ (1) 尾 _ (42), the viability of the cells was observed by MTT method, and the ideal A 尾 _ (1)-induced hippocampal neurons were selected as the cell model of Alzheimer's disease. The expression of neuron-specific enolase (NSE) in hippocampal neurons was determined by immunofluorescence (IF). Results the survival rate of hippocampal neurons induced by A 尾 _ (1) _ (42) decreased, which resulted in cell body deformation, contraction and cell membrane integrity destruction, disappearance or break of intercellular network structure and confusion of extracellular matrix. Neuronal apoptosis was observed in a large number of neurons. There was a linear relationship between A 尾 _ (1) _ (1) and A 尾 _ (1) ~ (42) concentration in hippocampal neurons after A 尾 _ (1) _ (42)-induced decrease in survival rate. After adding different concentrations of A 尾 _ (1) 42 (0 渭 mol / L, 10 渭 mol / L, 20 渭 mol / L, 30 渭 mol / L, 40 渭 mol / L and 50 渭 mol / L), the cell survival rates were (100.00 卤0.00)% and (98.55 卤1.42)%, respectively. (90.37 卤3.25)%, (66.34 卤2.76)%, (53.23 卤3.41)% and (41.55 卤2.37)%. P 0.05, 30 渭 mol / L, 40 渭 mol / L and 50 渭 mol / L in 20 渭 mol / L group were higher than those in control group (P 0.05, 30 渭 mol / L, 40 渭 mol / L and 50 渭 mol / L). Conclusion A 尾 _ (1) _ (42) can induce a large number of apoptosis of hippocampal neurons, and the hippocampal neurons induced by A 尾 _ (1) O _ (42) at 20 渭 mol / L can be used as an ideal cell model for Alzheimer's disease (AD) in SD rats.
【作者单位】：哈尔滨医科大学基础医学院形态实验中心;哈尔滨医科大学附属第五医院神经内科;
【分类号】：R749.16

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