中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (8): 1375-1383.doi: 10.4103/1673-5374.235250

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

染料木素对A53T突变α-核突触蛋白过表达SH-SY5Y细胞的神经保护

  

  • 收稿日期:2018-06-12 出版日期:2018-08-15 发布日期:2018-08-15
  • 基金资助:

    国家重点研究与发展计划(2016YFC1101500)；国家自然科学基金(11672332，11102235，8167050417)；天津市重点科技支持基金(17YFZCSY 00620)；天津市自然科学基金(15JCYBJC28600，17JCZDJC 35400)

Neuroprotective effects of genistein on SH-SY5Y cells overexpressing A53T mutant α-synuclein

Huan-Cheng Wu1, 2, Qun-Liang Hu2, Shi-Jun Zhang2, Yan-Min Wang2, Zhan-Kui Jin2, Ling-Fu Lv2, Sai Zhang3, Zhen-Lin Liu3,Hong-Lian Wu4, Ou-Mei Cheng4   

  1. 1 Graduate School, Tianjin Medical University, Tianjin, China
    2 Tianjin Beichen Hospital, Tianjin, China
    3 Tianjin Key Laboratory of Neurotrauma Repair, Institute of Traumatic Brain Injury and Neuroscience, Center for Neurology and Neurosurgery of Affiliated Hospital, Logistics University of Chinese People’s Armed Police Force, Tianjin, China
    4 Department of Clinical Medicine, Chongqing Medical University, Chongqing, China
  • Received:2018-06-12 Online:2018-08-15 Published:2018-08-15
  • Contact: Sai Zhang, M.D. or Zhen-Lin Liu, M.D.,zhangsai718@vip.126.com or wjzhenlin817@163.com.
  • Supported by:

    This study was supported by a grant from the National Key Research and Development Plan of China, No. 2016YFC1101500; the National Natural Science Foundation of China, No. 11672332, 11102235, 8167050417; the Key Science and Technology Support Foundation of Tianjin City of China, No. 17YFZCSY00620; the Natural Science Foundation of Tianjin City of China, No. 15JCYBJC28600, 17JCZDJC35400.

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摘要：

 

染料木素(GS)是一种有效的抗氧化剂化合物，已经被证实可以在帕金森病模型小鼠中保护多巴胺能神经元损伤，然而GS的神经保护机制尚不明确。为此，实验设计采用过表达A53T突变α-核突触蛋白的SH-SY5Y细胞为对象，分4组进行干预，对照组(不做任何干预)，GS组(单用GS，20 μM预处理)，鱼藤酮组(单用R 50 μM进行处理)及实验组(GS+R)。(1)实验通过乳酸脱氢酶释放试验证实了GS的保护作用，并发现GS能明显逆转鱼藤酮引起的线粒体氧化损伤；(2)以Western Blot检测发现，实验组抗凋亡蛋白Bcl-2和Beclin-1水平显著高于鱼藤酮(R)组，且TUNEL染色结果显示，GS对鱼藤酮诱导的SH-SY5Y细胞凋亡有抑制作用；(3)实验组细胞存活相关蛋白Nrf2和其下游蛋白HO-1表达显著增加，在使用Nrf2通道阻滞剂ML385干预后，GS并不能使Nrf2和HO-1蛋白表达增加；而雌激素受体抑制剂ICI-182780可有效地阻止GS介导的核Nrf 2的磷酸化，还能显著抑制雌激素受体下游通路蛋白AKT的磷酸化；(4)上述数据说明GS对帕金森病细胞模型有神经保护作用，它可以通过激活雌激素受体受体及Nrf2通道，减轻氧化应激损伤及细胞凋亡。

orcid:0000-0003-1974-079X(Zhen-Lin Liu)
        0000-0002-8028-4183(Sai Zhang)

关键词: 染料木素, 神经保护, SH-SY5Y细胞, 帕金森病, 鱼藤酮, 雌激素受体, Nrf2, A53T, &alpha, -核突触蛋白, 氧化应激, 神经退行性变, 神经再生

Abstract:

Genistein, a potent antioxidant compound, protects dopaminergic neurons in a mouse model of Parkinson’s disease. However, the mechanism underlying this action remains unknown. This study investigated human SH-SY5Y cells overexpressing the A53T mutant of α-synuclein.Four groups of cells were assayed: a control group (without any treatment), a genistein group (incubated with 20 μM genistein), a rotenone group (treated with 50 μM rotenone), and a rotenone + genistein group (incubated with 20 μM genistein and then treated with 50 μM rotenone). A lactate dehydrogenase release test confirmed the protective effect of genistein, and genistein remarkably reversed mitochondrial oxidative injury caused by rotenone. Western blot assays showed that BCL-2 and Beclin 1 levels were markedly higher in the genistein group than in the rotenone group. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling revealed that genistein inhibited rotenone-induced apoptosis in SH-SY5Y cells. Compared with the control group, the expression of NFE2L2 and HMOX1 was significantly increased in the genistein + rotenone group. However, after treatment with estrogen receptor and NFE2L2 channel blockers (ICI-182780 and ML385, respectively), genistein could not elevate NFE2L2 and HMOX1 expression. ICI-182780 effectively prevented genistein-mediated phosphorylation of NFE2L2 and remarkably suppressed phosphorylation of AKT, a protein downstream of the estrogen receptor. These findings confirm that genistein has neuroprotective effects in a cell model of Parkinson’s disease. Genistein can reduce oxidative stress damage and cell apoptosis by activating estrogen receptors and NFE2L2 channels.

Key words: nerve regeneration, genistein, neuroprotection, SH-SY5Y cells, Parkinson’s disease, rotenone, estrogen receptor, NFE2L2, A53T, α-synuclein, oxidative stress, neurodegeneration, neural regeneration