解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (5): 616-621.doi: 10.3969/j.issn.0529-1356.2014.05.006

• 神经生物学 • 上一篇    下一篇

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1,2 郑关毅1* 陈晓春1 张静1 黄天文1 叶洪1 潘晓东1   

  1. 1. 福建医科大学附属协和医院中医科 福建省老年医学研究所，福州 350001; 2. 江苏省沭阳县人民医院神经内科，江苏 沭阳 223600
  • 收稿日期:2013-12-09 修回日期:2014-03-17 出版日期:2014-10-06 发布日期:2013-10-06
  • 通讯作者: 郑关毅 E-mail:ycyz99@163.com
  • 基金资助:

    福建省卫生厅中医药重点课题;福建省自然科学基金资助项目

Ativation of gliacytes and p38 mitogen-activated protein kinase and possible mechanism of neuronal apoptosis induced by Aβ25-35 injection into hippocampus in rats

WANG Yuan-wei 1,2 ZHENG Guan-yi 1* CHEN Xiao-chun1 ZHANG Jing1 HUANG Tian-wen1 YE Hong1 PAN Xiao-dong1   

  1. 1. Department of Traditional Chinese Medicine,Fujian Medical University Union Hospital, Geriatric Research Institute of Fujian Province, Fuzhou 350001, China; 2. Department of Neurology, Shuyang People’s Hospital of Jiangsu Province, Jiangsu Shuyang 223600, China
  • Received:2013-12-09 Revised:2014-03-17 Online:2014-10-06 Published:2013-10-06
  • Contact: ZHENG Guan-yi E-mail:ycyz99@163.com

摘要：

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。

关键词: 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

Abstract:

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.

Key words: Alzheimer’s disease, Amyloid protein β, P38 mitogen-activated protein kinases, Gliacyte, Hippocampal neuron,  , Wstern blotting, Rat