中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (12): 2296-2305.doi: 10.4103/1673-5374.285006

• 原著:退行性病与再生 • 上一篇    下一篇

ZL006减轻Aβ1-42诱导神经元损伤的作用途径

  


  • 出版日期:2020-12-15 发布日期:2020-08-05
  • 基金资助:
    国家自然科学基金面上项目（81671055、81971009），中国江苏省重点研究发展计划（BE2016610），中国国家重点研究开发计划（2016YFC1300504，2018YFC1704405），江苏省重点医学学科项目（ZDXKA2016020），江苏省青年医学人才项目（QNRC2016024），青年人才支持计划江苏省科学技术协会项目。

Neuroprotective efects of ZL006 in Aβ1–42-treated neuronal cells

Wen-Yuan Tao 1, 2, 3, Lin-Jie Yu 1, 2, 3 , Su Jiang 4 , Xiang Cao 1, 2, 3 , Jian Chen 1, 2, 3 , Xin-Yu Bao 1, 2, 3 , Fei Li 5 , Yun Xu 1, 2, 3 , Xiao-Lei Zhu 1, 2, 3   

  1. 1 Department of Neurology, Drum Tower Hospital, Medical School of Nanjing University, Nanjing, Jiangsu Province, China
    2 The State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, Jiangsu Province, China
    3 Jiangsu Key Laboratory for Molecular Medicine, Nanjing, Jiangsu Province, China
    4 Taizhou People’s Hospital, Taizhou, Jiangsu Province, China
    5 Department of Medicinal Chemistry, School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu Province, China
  • Online:2020-12-15 Published:2020-08-05
  • Contact: Xiao-Lei Zhu, MD, PhD,zhuquelee@126.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, Nos. 81671055, 81971009 (to XLZ), the Key Research and Development Program of Jiangsu Province of China, No. BE2016610 (to YX), the National Key Research and Development Program of China, No. 2016YFC1300504 (to YX) and No. 2018YFC1704405 (to XLZ), Jiangsu Province Key Medical Discipline, No. ZDXKA2016020 (to YX), Jiangsu Province Medical Youth Talent, No. QNRC2016024 (to XLZ), and Young Talent Support Program from Jiangsu Association for Science and Technology (to XLZ).

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摘要：

β淀粉样蛋白(amyloid beta，Aβ)诱导的神经毒性和氧化应激在阿尔茨海默病的发生和发展中起重要作用。既往已经证明 ZL006可阻断缺血性卒中时神经元一氧化氮合酶与突触后密度蛋白95的相互作用，可降低氧化应激水平发挥神经保护作用。关于ZL006在阿尔茨海默病中的作用报道较少。假设ZL006在阿尔茨海默病中具有神经保护作用，那么可能的机制是什么？实验采用Aβ1-42处理原代大脑皮质神经元和N2a神经母细胞瘤细胞作为阿尔茨海默病的体外模型，用ZL006或DMSO预处理大脑皮质神经元2 h后，再用Aβ1-42或氨水处理24 h。(1) 通过CCK8和calcein-AM/PI实验发现，ZL006预处理能挽救Aβ1-42诱导的大脑皮质神经元死亡；(2)通过免疫荧光和蛋白免疫印记等方法检测到，细胞氧化应激和凋亡相关指标的变化，在 ZL006预处理后能激活大脑皮质神经元内Akt/Nrf2/HO-1信号通路并减少Aβ1-42诱导的神经元凋亡和氧化应激；(3)CCK8实验发现，抑制大脑皮质神经元内Akt或Nrf2则阻断了ZL006的神经保护作用；(4)在N2a细胞中也能观察到类似结果，ZL006能够抑制Aβ1-42诱导的N2a细胞损伤及氧化应激，而抑制Akt或Nrf2能够部分阻断ZL006的保护作用；(5)这些结果可验证，ZL006减轻了Aβ1-42诱导的神经元损伤和氧化应激，作用途径在与激活了Akt/Nrf2/HO-1信号通路。

orcid:0000-0003-4696-9696(Xiao-Lei Zhu)

关键词: 阿尔茨海默氏病, 细胞凋亡 Akt, β淀粉样蛋白 HO-1, 神经毒性, Nrf2, 氧化应激治疗, ZL006

Abstract: Amyloid beta (Aβ)-induced neurotoxicity and oxidative stress plays an important role in the pathogenesis of Alzheimer’s disease (AD). ZL006 is shown to reduce over-produced nitric oxide and oxidative stress in ischemic stroke by interrupting the interaction of neuronal nitric oxide synthase and postsynaptic density protein 95. However, few studies are reported on the role of ZL006 in AD. To investigate whether ZL006 exerted neuroprotective effects in AD, we used Aβ 1–42 to treat primary cortical neurons and N2a neuroblastoma cells as an in vitro model of AD. Cortical neurons were incubated with ZL006 or dimethyl sulfoxide for 2 hours and treated with Aβ 1–42 or NH3•H 2 O for another 24 hours. The results of cell counting Kit-8 (CCK-8) assay and calcein-acetoxymethylester/propidium iodide staining showed that ZL006 pretreatment rescued the neuronal death induced by Aβ 1–42 . Fluorescence and western blot assay were used to detect oxidative stress and apoptosis-related proteins in each group of cells. Results showed that ZL006 pretreatment decreased neuronal apoptosis and oxidative stress induced by Aβ 1–42 . The results of CCK8 assay showed that inhibition of Akt or NF-E2-related factor 2 (Nrf2) in cortical neu- rons abolished the protective effects of ZL006. Moreover, similar results were also observed in N2a neuroblastoma cells. ZL006 inhibited N2a cell death and oxidative stress induced by Aβ 1–42 , while inhibition of Akt or Nrf2 abolished the protective effect of ZL006. These results demonstrated that ZL006 reduced Aβ 1–42 -induced neuronal damage and oxidative stress, and the mechanisms might be associated with the activation of Akt/Nrf2/heme oxygenase-1 signaling pathways.

Key words: Akt, Alzheimer’s disease, amyloid-beta, apoptosis, heme oxygenase-1, neurotoxicity, Nrf2, oxidative stress, treatment, ZL006