β淀粉样蛋白25~35 对PC12细胞骨架的毒性作用

doi:10.16098/j.issn.0529-1356.2016.04.06

摘要/Abstract

摘要：

目的建立阿尔茨海默病（AD）的细胞模型，并探讨β淀粉样蛋白25~35（Aβ _25~35）对细胞骨架的神经毒性机制。方法采用MTT法检测不同浓度的Aβ _25~35 对PC12细胞存活率的影响，应用4’6-二脒基-2-苯基吲哚（DAPI）染色法和TUNEL法检测细胞凋亡，免疫细胞化学法和鬼笔环肽（phalloidin）染色观察细胞骨架的形态变化。结果 Aβ _25~35经过“老化”处理后，可以明显引发PC12细胞死亡，导致PC12细胞发生凋亡，并具有剂量依赖性（P<0.05）。Aβ _25~35也可以引起细胞骨架解体，同样具有剂量依赖性（P<0.05）。结论 PC12细胞的细胞骨架对Aβ25~35的神经毒性非常敏感，细胞骨架的解体很可能是AD重要的病理学改变，同时Aβ是AD发病机制的关键分子。

关键词: 阿尔茨海默病, β淀粉样蛋白_25~35, 细胞骨架, 原位缺口末端标记, 免疫组织化学, PC12细胞

Abstract:

Objective To establish the cell model of Alzheimer’s disease (AD) and investigate the amyloid beta-peptides _25-35(Aβ _25-35) neurotoxicity to cytoskeleton.Methods PC12 cells were cultured and treated with Aβ _25-35, and cell survival was analyzed by MTT assay. Cell apoptosis was visualized with DAPI staining and TUNEL method. Immunocytochemistry and phalloidin staining were used to label cytoskeletons in PC12 cells. Results Aβ _25-35 obviously induced the PC12 cells death and lead to PC12 cells apoptosis with dose dependency (P<0.05). Aβ _25-35 gave rise to the disintegration of cytoskeletons with dose dependency (P<0.05). Conclusion PC12 cell cytoskeletons are sensitive to Aβ _25-35 neurotoxicity. The disintegration of cytoskeleton probably is the important pathological alteration in AD, and Aβ is a key molecule for AD pathogenesis.

Key words: Alzheimer’s disease, Amyloid beta-peptides 25-35, Cytoskeleton, Terminal UTP nick end labeling, Immunohistochemistry, PC12 cell

曹晶晶范文娟石贞玉鄢明超刘红亮王来邓锦波. β淀粉样蛋白25~35 对PC12细胞骨架的毒性作用[J]. 解剖学报, 2016, 47(4): 469-475.

CAO Jing-jing FAN Wen-juan SHI Zhen-yu YAN Ming-chao LIU Hong-liang WANG Lai DENG Jin-bo. Effect of amyloid beta-peptide 25-35 neurotoxicity on cytoskeletons of PC12 cells[J]. Acta Anatomica Sinica, 2016, 47(4): 469-475.

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