Aβ_(25-35)寡聚体对大鼠海马神经细胞的活性影响及形态学观察

The effect of Aβ_(25-35) oligomer on survival and morphology of primary cultured hippocampal neurons survival and morphology

目的研究以β淀粉样蛋白(Aβ25-35)损伤原代培养海马神经元建立Alzheimer病(AD)细胞模型的方法。方法运用细胞原代培养的方法培养大鼠海马神经元并进行鉴定,以不同浓度Aβ25-35寡聚体建立海马神经元损伤模型,将培养的细胞分为Aβ25-35寡聚体致伤高、中、低剂量组,同时设立正常对照组,倒置显微镜观察细胞形态学变化及通过四唑盐(MTT)比色实验检测细胞存活率。结果当Aβ25-35寡聚体终浓度为5.0μmol/L、10.0μmol/L、20.0μmol/L时,作用于细胞24h,可使神经细胞的形态发生改变和活力显著下降,在显微镜下可见神经元形态有明显的变化,失去贴壁能力或易脱落、突起变短、细胞存活率明显下降,与对照组相比较差异有显著性(P<0.01)。结论 Aβ25-35寡聚体可导致原代培养海马神经元变性、死亡,且有剂量依赖关系,可用于构建AD的细胞模型;并筛选出5.0μmol/L Aβ25-35寡聚体为AD模型的适宜致伤浓度。

Objective To study establishing Alzheimer disease （AD）cell model by Aβ_25-35 oligomer induced injury of primary cultured hippocampal neurons. Methods Hippoeampal neurons cell derived from rats by primary culture meth- od wereas identified, and hippocampal neuron injury model was established with different concentrations of Aβ_25-35 oli- gomer. All Ccultured cells were divided into 5 groups ,with including 1.0μmol/L,5.0μ moL/L, 10.0μ mol/L,20.0μ mol/ L group Aβ_25-35 oligomer per group, and control group. Then Mmorphological changes were observed by inverted microscope and survival rate was assayed by MTF. Results compared with the control group, Aβ_25-35 oligomer with the concentra- tions of 5.0μmol/L, 10.0μmol/L and,20.0p, mol/L for 24h, can c ausedmake the nerve cell morphological changes and vigor decreased vigor significantly. Survival of Ceells with Aβ_25-35 oligomer survival decreased significantly compared with the control group （P 〈 0.01 ）. Conclusion Aβ_25-35 oligomer dose-dependently can result inused primary cultured hipp- ocampal neuronal degeneration, and death in primary cultured hippocampal neuron, and there is a dose-dependent relation- ship. Aβ_25-35 oligomer can be used to establish AD cell model,and 5.0μ mol/L is a properprefer concentration.