Beta-amyloid toxicity in embryonic rat astrocytes
Beta-amyloid toxicity in embryonic rat astrocytes
复制标题
DOI：
10.1007/s11064-007-9335-8
复制
发表时间：
2007-09-01
影响因子：
4.4
通讯作者：
Mudd, Laura M.
中科院分区：
文献类型：
--
作者：
Assis-Nascimento, Poincyane;Jarvis, Karen M.;Mudd, Laura M.
阿尔茨海默病的老年斑含有高浓度的β-淀粉样蛋白（β A），这可能会影响隔核中的神经胶质细胞群，这是AD风险增加的区域。通过剂量反应实验，通过定量三种不同剂量（0.1、1和10 μ M）的β A对细胞存活的影响，在隔神经胶质中测量β A毒性。来自胚胎第16天大鼠的星形胶质细胞在无血清培养基中单层培养。在体外第1天（DIV）处理细胞，并在DIV 3测定存活率，以确定哪个浓度毒性最大。在一组单独的实验中，试图通过生长因子和雌激素的治疗来保护神经胶质细胞免受β A的退行性作用。在DIV 1时给予β A（10 μ M）处理，在DIV 2时用雌激素处理细胞（EST，10 nM），胰岛素样生长因子（IGF 1和IGF 2，各10 ng/ml）、碱性成纤维细胞生长因子（bFGF，5 ng/ml）或神经生长因子（NGF，100 ng/ml），并且在DIV 3上，通过荧光显微术用DAPI（4，6-二脒基-2-苯基吲哚）使细胞可视化并定量。除了剂量反应和神经胶质保护外，还进行了实验以确定毒性作用是否归因于细胞凋亡。我们的研究结果表明，神经胶质细胞群的生存显着影响在所有三个浓度（0.1，1.0，和10 μ M）的β A。神经胶质保护在NGF存在下是明显的，因为它显示出相对于单独的β A治疗的显著最高的存活率。此外，β A的毒性作用似乎主要是由于细胞凋亡。bFGF和IGF 1可显著逆转β A诱导的细胞凋亡。
The senile plaques of Alzheimer's disease contain a high concentration of beta-amyloid (beta A) protein, which may affect the glial population in the septal nucleus, an area of increased risk in AD. beta A toxicity was measured in septal glia, via a dose-response experiment, by quantifying the effects of three different doses (0.1, 1, and 10 mu M) of beta A on cell survival. Astrocytes from embryonic day-16 rats were grown in serum-free media in a single layer culture. Cells were treated on day in vitro (DIV)1 and survival was determined on DIV3 to ascertain which concentration was most toxic. In a separate set of experiments, an attempt was made to protect glial cells from the degenerative effects of beta A, with treatments of growth factors and estrogen. beta A (10 mu M) treatment was administered on DIV1, on DIV2 the cells were treated with estrogen (EST, 10 nM), insulin-like growth factors (IGF1 and IGF2, each 10 ng/ml), basic fibroblast growth factor (bFGF, 5 ng/ml) or nerve growth factor (NGF, 100 ng/ml), and on DIV3 the cells were visualized and quantified by fluorescence microscopy with DAPI (4,6-diamidino-2-phenylindole). In addition to dose-response and glial protection, experiments were also conducted to determine whether toxic effects were due to apoptosis. Our results suggest that the survival of glial populations is significantly affected in all three concentrations (0.1, 1.0, and 10 mu M) of beta A. Glial protection was evident in the presence of NGF, for it showed the significantly highest survival rate relative to the beta A treatment alone. Furthermore, toxic effects of beta A appear to be due primarily to apoptosis. Significant reversal of beta A-induced apoptosis was seen with bFGF and IGF1.