星形胶质细胞对天冬氨酸特异性半胱氨酸蛋白酶介导β淀粉样蛋白早期突触毒性作用的影响

中国卒中杂志 ›› 2015, Vol. 10 ›› Issue (03): 225-230.

星形胶质细胞对天冬氨酸特异性半胱氨酸蛋白酶介导β淀粉样蛋白早期突触毒性作用的影响

吕昂初1，宋一志1，张亚丽1，李彦1，方圆1，陆涛1，刘津平2，常丽荣1，武艳1

1100069 北京
首都医科大学人体解剖与组织胚胎学系
2清华大学医学院

收稿日期:2013-09-11 出版日期:2015-03-20 发布日期:2015-03-20
通讯作者: 武艳 yanwu@ccmu.edu.cn
基金资助:
北京市自然科学基金面上项目（5152004）
北京市属高等学校高层次人才引进与培养计划项目-青年拔尖人才（11350901）
新世纪优秀人才支持计划资助（NCET-10-0015）
北京教委科技发展计划
（SQKM201210025003）
国家自然科学基金
（81301100，81200968）

Effect of Astrocytes on Caspase-mediated Early Aβ Synaptotoxicity

*Department of Anatomy, Capital Medical University, Beijing 100069, China

Received:2013-09-11 Online:2015-03-20 Published:2015-03-20

摘要/Abstract

摘要：

目的探讨星形胶质细胞（astrocyte，AS）对天冬氨酸特异性半胱氨酸蛋白酶（cysteinyl aspartate specific proteinase，caspase）介导β淀粉样蛋白（β-amyloid，Aβ）早期突触毒性作用的影响，以期为进一步研究与血管性痴呆（vascular dementia，VaD）的发病机制奠定基础。方法以原代培养大鼠海马纯神经元体系（NE-S）及混合培养体系（MIX-S，主要包含神经元及AS）为研究对象，各体系分为6组：对照组、caspase-8抑制剂组、caspase-9抑制剂组、Aβ处理组、caspase-8抑制剂预处理加Aβ组和caspase-9抑制剂预处理加Aβ组。免疫荧光检测各组近胞体10?μm段树突中突触后密度蛋白（postsynaptic density-95，PSD95）表达量的变化。结果 ①在NE-S与MIX-S中，与对照组相比，caspase-8抑制剂组、caspase-9抑制剂组PSD95的表达量均无明显差异，Aβ处理组PSD95的表达量均显著降低（P均＜0.001）。②在NE-S中，与Aβ处理组相比，caspase-9抑制剂预处理加Aβ组PSD95的表达量显著回升至对照组水平，caspase-8抑制剂预处理加Aβ组则无显著改变；在MIX-S中的结果则相反，即caspase-8抑制剂预处理加Aβ组PSD95的表达量显著回升至对照组水平，而caspase-9抑制剂预处理加Aβ组则无显著改变。③MIX-S与NE-S两种培养系统间相比较，对照组间及Aβ处理组间PSD95的表达量均无显著差异，而caspase-8抑制剂预处理加Aβ组间及caspase-9抑制剂预处理加Aβ组间PSD95的表达量差异有显著性。结论在Aβ早期突触毒性作用中，AS参与caspase-8介导的死亡受体通路激活过程，且参与抑制神经元的线粒体通路。

文章导读： 对星形胶质细胞在β淀粉样蛋白早期突触毒性作用的影响机制进行深入研究将为发现阿尔茨海默病与血管性痴呆的早期诊断、治疗的新靶点提供重要的理论依据。

关键词: 血管性痴呆；星形胶质细胞；β 淀粉样蛋白；天冬氨酸特异性半胱氨酸蛋白酶；突触

Abstract:

Objective To investigate the effect of astrocytes on caspase-mediated early β-amyloid (Aβ) neurotoxicity so as to lay a foundation for further studying of the pathogenesis of vascular dementia (VaD). Methods We established the 4-day-old Wistar rat hippocampal primary mixed culture system (MIX-S), neurons and astrocytes are mainly included, and the purified primary neuronal culture system (NE-S). Then each system was divided into six groups: control group, caspase-8 inhibitor group, caspase-9 inhibitor group, Aβ group, caspase-8 inhibitor pretreatment with Aβ group and caspase-9 inhibitor pretreatment with Aβ group. Immunofluorescence technique was applied to investigate the changes of the expression of postsynaptic density-95 (PSD95) at 10?μm dendrites near to cell body. Results ① Both in NE-S and MIX-S, compared with control group, the expression of PSD95 had no significant difference in the caspase-8 inhibitor group and the caspase-9 inhibitor group, while that in the Aβ group reduced significantly. ② In NE-S, compared with the Aβ group, the expression of PSD95 rebounded significantly to the level of the control group in the caspase-9 inhibitor pretreatment with Aβ group, while no significant change was found in the caspase-8 inhibitor pretreatment with Aβ group; interestingly, the results of this part in MIX-S was reversed. ③ Compared with NE-S, the expression of PSD95 had no significant change in the control group and Aβ group of MIX-S; while the difference of that was significant in the caspase-8 inhibitor pretreatment with Aβ group and caspase-9 inhibitor pretreatment with Aβ group. Conclusion Astrocyte plays a role in activation of caspase-8 mediated death receptor pathway, and also participates in inhibition of neuronal mitochondrial pathway in early Aβ synaptotoxicity.

Key words: Vascular dementia; Astrocytes; β-amyloid; Caspase; Synapse

吕昂初1，宋一志1，张亚丽1，李彦1，方圆1，陆涛1，刘津平2，常丽荣1，武艳1. 星形胶质细胞对天冬氨酸特异性半胱氨酸蛋白酶介导β淀粉样蛋白早期突触毒性作用的影响[J]. 中国卒中杂志, 2015, 10(03): 225-230.

LV Ang-Chu*, SONG Yi-Zhi, ZHANG Ya-Li, LI Yan, FANG Yuan, LU Tao, LIU Jin-Ping, CHANG Li-Rong, WU Yan. . Effect of Astrocytes on Caspase-mediated Early Aβ Synaptotoxicity[J]. Chinese Journal of Stroke, 2015, 10(03): 225-230.

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