上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2018, Vol. 38 ›› Issue (9): 1013-.doi: 10.3969/j.issn.1674-8115.2018.09.003

• 论著·基础研究 • 上一篇    下一篇

Rho相关的卷曲蛋白激酶 1参与 β淀粉样蛋白介导的大鼠原代神经元损伤

胡勇博 1, 2，任汝静 1，王刚 1   

  1. 1. 上海交通大学医学院附属瑞金医院神经内科，上海 200025；2. 上海交通大学基础医学院药理学与化学生物学系，上海 200025
  • 出版日期:2018-09-28 发布日期:2018-10-15
  • 通讯作者: 王刚，电子信箱：wg11424@rjh.com.cn。
  • 作者简介:胡勇博，（1991—），男，博士生；电子信箱： huyongbo91@126.com。
  • 基金资助:
    国家自然科学基金（ 81671043）；上海市浦江人才计划（ 15PJ1405400）；上海市曙光计划（ 16SG15）；上海市教育委员会高峰高原项目（ 20172001）

Rho-associated coiled coil protein kinase 1 involved in amyloid β-protein-induced damage in primary neurons of rats

HU Yong-Bo1,2, REN Ru-Jing1, WANG Gang1   

  1. 1. Department of Neurology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 2. Department of Pharmacology and Chemical Biology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China
  • Online:2018-09-28 Published:2018-10-15
  • Supported by:
    National Natural Science Foundation of China, 81671043; Shanghai Pujiang Program, 15PJ1405400; the "Dawn" Program of Shanghai Education Commission, 16SG15; Shanghai Municipal Education Commission— Gaofeng Clinical Medicine Support, 20172001

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摘要： 目的 ·探讨 Rho相关的卷曲蛋白激酶 1（Rho-associated coiled coil protein kinase 1，ROCK1）是否参与了 β淀粉样蛋白（amyloid β-protein，Aβ）引起大鼠海马神经元损伤的过程。方法 ·用 Aβ40寡聚肽处理大鼠海马原代神经元，建立神经元损伤模型。利用免疫印迹法检测 ROCK1蛋白表达水平，试剂盒检测 ROCK1激酶活性，激光共聚焦显微镜检测荧光染料 Fluo-8AM标记的细胞内钙离子负荷， TUNEL染色检测细胞凋亡情况。加入 ROCK1抑制剂 Y-27632，观察其对 Aβ40效应的影响。结果 · 10 μmol/L Aβ40寡聚肽能引起神经元内钙超载， ROCK1蛋白表达增加和活性升高，以及细胞凋亡比例升高；而 Y-27632能对抗 Aβ40引起的这些效应。结论 · ROCK1参与了 Aβ40诱导的神经元内钙超载及神经毒性的产生，而 ROCK1抑制剂能拮抗 Aβ毒性效应。

关键词: 阿尔茨海默病, Rho相关的卷曲蛋白激酶 1, &, beta, 淀粉样蛋白, 钙超载

Abstract:

Objective · To investigate the role of Rho-associated coiled coil protein kinase 1 (ROCK1) in amyloid β-protein (Aβ) induced damage of rat hippocampal neurons. Methods · The rat primary neurons were treated with Aβ40 oligopeptides to establish a neurotoxicity model. Western blotting was used to detect the protein of ROCK1. Its activity was detectedthe kit. Confocal laser scanning was used to observe the calcium signal in neurons, and apoptosis of neurons was detectedTUNEL assay. Y-27632, an inhibitor of ROCK1, was added into the culture medium into observe its effect on Aβ40. Results · Aβ40 (10 μmol/L) could significantly induce calcium overload, increase ROCK1 and activity, and promote apoptosis in primary neurons. Furthermore, ROCK1 inhibitor could decrease all the effect inducedAβ40. Conclusion · ROCK1 is involved in both Aβinduced neuronal calcium overload and neurotoxicity, and ROCK1 inhibitor can antagonize the toxic effects of Aβ.

Key words: Alzheimers disease, Rho-associated coiled coil protein kinase 1 (ROCK1), amyloid β-protein (Aβ), calcium overload

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