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海马背侧注射β-淀粉样蛋白 25~35 激活胶质细胞和p38丝裂原活化蛋白激酶诱导神经元凋亡
王元伟 郑关毅* 陈晓春 张静 黄天文 叶洪 潘晓东
解剖学报 ›› 2014, Vol. 45 ›› Issue (5) : 616-621.
海马背侧注射β-淀粉样蛋白 25~35 激活胶质细胞和p38丝裂原活化蛋白激酶诱导神经元凋亡
Ativation of gliacytes and p38 mitogen-activated protein kinase and possible mechanism of neuronal apoptosis induced by Aβ25-35 injection into hippocampus in rats
目的 探讨海马背侧注射β-淀粉样蛋白25～35（Aβ 25～35）后胶质细胞与 p38丝裂原活化蛋白激酶（p38MAPK）激活的关系，及Aβ 25～35 诱导神经元凋亡的可能机制。 方法 采用免疫组织化学及免疫印迹方法观察海马背侧注射Aβ 25～35 后不同时段小胶质细胞（MG）、星形胶质细胞（AS）的激活和磷酸化p38MAPK（p-p38MAPK）在海马组织中的表达；酶联免疫吸附法检测肿瘤坏死因子-α（TNF-α）及白细胞介素-1β（IL-1β）在海马组织中的含量；Nissl染色法观察海马神经元存活；TUNEL染色观察海马神经元凋亡。 结果 注射Aβ 25～35 后海马内抗特异性标记小胶质细胞(ox-42)、胶质原纤维酸性蛋白(GFAP)、和p-p38MAPK的表达与TNF-α、IL-1β的含量同步增加，于7d达到高峰，而Nissl阳性神经元数量逐渐减少，TUNEL阳性神经元数量则逐渐增多，亦于7d达到高峰。结论 海马背侧注射Aβ 25～35 可能通过激活胶质细胞和p38MAPK，使TNF-α，IL-1β含量增加导致海马神经元凋亡。
Objective To investigate the relationship between activation of gliacytes, mitogen-activated protein kinase (p38MAPK) and neuronal apoptosis after microinjecting aggregated Aβ25-35 into hippocampus. Methods The model was established by using stereotaxic technique to inject 10μg aggregated Aβ25-35 into dorsal hippocampus in rats. The rats were grouped as the control, vehicle and model groups. Immunohistochemistry and Western blotting were used for detection of activation of microglia(MG), atrocytes (AS) and expression of p-p38MAPK in the hippocampus. ELISA was used to evaluate the level of TNF-α and IL-1β. The survival neurons were observed by Nissl staining and the apoptotic neurons were identified by tunnel staining. Results Expression of ox-42, GFAP, p-p38MAPK were up-regulated in hippocampus, as well as TNF-α、IL-1β, which reached a highest value on the 7th day after injection of Aβ25-35. However, the number of neuron with Nissl positive decreased gradually, and the tunnel positive neurons increased highly and reached a peak value on the 7th day.There were significant differences between the control and vehicle group（P<0.01）. Conclusion Apoptosis of the neuron caused by Aβ25-35 injection may result from activation of gliacytes, p38 MAPK and increase of TNF-α and IL-1β level.
阿尔茨海默病 / β-淀粉样蛋白 / p38丝裂原活化蛋白激酶 / 胶质细胞 / 海马神经元 / 免疫印迹法 / 大鼠
Alzheimer’s disease / Amyloid protein β / P38 mitogen-activated protein kinases / Gliacyte / Hippocampal neuron / / Wstern blotting / Rat
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