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 (9): 1552-1560.doi: 10.4103/1673-5374.235299

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

线粒体分裂抑制因子1保护大脑皮质神经元免于兴奋性毒性损伤的作用途径

  

  • 收稿日期:2018-06-16 出版日期:2018-09-15 发布日期:2018-09-15
  • 基金资助:

    中国国家自然科学基金项目（81371967，81401807），第5期333工程项目（BRA2016512），江苏省六大人才高峰资助项目（2014-WSN-012）

Mitochondrial division inhibitor 1 protects cortical neurons from excitotoxicity: a mechanistic pathway

Kuai Zhou1, Hai-Yuan Yang2, Peng-Yu Tang1, Wei Liu1, Yong-Jun Luo1, Bin Lv1, Jian Yin3, Tao Jiang1, Jian Chen1, Wei-Hua Cai1, Jin Fan1   

  1. 1 Department of Orthopedics, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China;
    2 Department of Orthopedics, BenQ Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu Province, China;
    3 Department of Orthopedics, Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
  • Received:2018-06-16 Online:2018-09-15 Published:2018-09-15
  • Contact: Wei-Hua Cai, Ph.D. or Jin Fan, Ph.D., caiwhspine@sina.com or fanjin@njmu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81371967 and 81401807; a grant from the 5th Phase of “Project 333” of Jiangsu Province of China, No. BRA2016512; and a grant from the Six Talent Peaks Project of Jiangsu Province of China, No. 2014-WSN-012.

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摘要：

线粒体分裂抑制因子1是动力相关蛋白1和线粒体分裂的选择性细胞渗透性抑制剂。为研究其对谷氨酸暴露大脑皮质神经细胞的影响，实验对分离的新生大鼠大脑皮质神经元加入10 mM谷氨酸干预24h，并且以正常培养的细胞以及以DMSO培养的细胞作为对照。结果以流式细胞术检测凋亡细胞，以电子显微镜观察线粒体形态变化，以western blot和免疫组化检测Drp1，Bax和caspase-3的蛋白表达及免疫反应，以JC-1探针检测线粒体膜电位。对于10 mM谷氨酸暴露24h的细胞，其Drp1，Bax和caspase-3表达上调，且 Drp1和Bax向线粒体转移，线粒体膜电位下降，凋亡增加；而这些效应在经50 μM Mdivi-1预处理2h后就会被抑制；提示作为一种潜在的神经保护药物，线粒体分裂抑制因子1具有减轻由谷氨酸诱导的神经元兴奋性毒性损伤的潜力。

orcid:0000-0002-8923-3948(Wei-Hua Cai)

关键词: 神经再生, 线粒体分裂抑制因子1, 神经元, 细胞凋亡, 线粒体, 细胞分裂, 动力相关蛋白1, 磷酸化动力相关蛋白1, Bax, 谷氨酸, 共定位

Abstract:

Mitochondrial division inhibitor 1 (Mdivi-1) is a selective cell-permeable inhibitor of dynamin-related protein-1 (Drp1) and mitochondrial division. To investigate the effect of Mdivi-1 on cells treated with glutamate, cerebral cortex neurons isolated from neonatal rats were treated with 10 mM glutamate for 24 hours. Normal cultured cells and dimethyl sulfoxide-cultured cells were considered as controls. Apoptotic cells were detected by flow cytometry. Changes in mitochondrial morphology were examined by electron microscopy. Drp1, Bax, and casp ase-3 expression was evaluated by western blot assays and immunocytochemistry. Mitochondrial membrane potential was detected using the JC-1 probe. Twenty-four hours after 10 mM glutamate treatment, Drp1, Bax and caspase-3 expression was upregulated, Drp1 and Bax were translocated to mitochondria, mitochondrial membrane potential was decreased and the rate of apoptosis was increased. These effects were inhibited by treatment with 50 μM Mdivi-1 for 2 hours. This finding indicates that Mdivi-1 is a candidate neuroprotective drug that can potentially mitigate against neuronal injury caused by glutamate-induced excitotoxicity.

Key words: nerve regeneration, mitochondrial division inhibitor 1, neurons, apoptosis, mitochondria, division, dynamin-related protein-1, phospho-dynamin-related protein-1, Bax, glutamate, colocalization, neural regeneration