依达拉奉对MPP⁺处理的PC12细胞线粒体融合和分裂平衡的保护作用
焦阳¹, 郑月², 宋成洁^3,*
1中国人民解放军陆军第七十一集团军医院，徐州 221004；²南京医科大学生物化学与分子生物学系，南京 210029；³徐州医 科大学生理学教研室，徐州 221004
摘要
本文旨在探讨依达拉奉(edaravone, Eda)对帕金森病细胞模型线粒体融合、分裂动态平衡的作用及机制。用500 μmol/L 1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium, MPP+)处理PC12细胞建立帕金森病细胞模型，采用噻唑蓝(MTT)比色法 检测不同浓度Eda对MPP+处理的PC12细胞存活率的影响，用激光共聚焦显微镜检测线粒体形态，用Western blot检测线粒体融 合与分裂相关蛋白OPA1、MFN2、DRP1和Fis1的表达变化。结果显示，预先加入不同浓度的Eda能减轻MPP+处理的PC12细胞 损伤，作用呈一定的量效关系；经MPP+处理48 h，PC12细胞线粒体出现碎片化，OPA1和MFN2蛋白表达下调，DRP1和Fis1 蛋白表达上调，而Eda预处理能逆转PC12细胞的上述变化，但对Fis1的蛋白表达没有影响。以上结果提示，Eda可上调OPA1 和MFN2的蛋白表达，下调DRP1的表达，从而抑制线粒体碎片化，发挥神经细胞线粒体保护作用。


关键词： 依达拉奉; MPP+; PC12细胞; 线粒体融合; 线粒体分裂
分类号：R285.5

Protective effect of edaravone on balance of mitochondrial fusion and fission in MPP⁺-treated PC12 cells
JIAO Yang¹, ZHENG Yue², SONG Cheng-Jie^3,*
1The 71st Group Military Hospital of the Chinese People’s Liberation Army, Xuzhou 221004, China；²Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing 210029, China；³Department of Physiology, Xuzhou Medical University, Xuzhou 221004, China
Abstract
The aim of this study was to investigate the effect of edaravone (Eda) on the balance of mitochondrial fusion and fission in Parkinson’s disease (PD) cell model. A cell model of PD was established by treating PC12 cells with 500 μmol/L 1-methyl-4-phenylpyridinium (MPP+). Thiazole blue colorimetry (MTT) was used to detect the effect of different concentrations of Eda on the survival rate of PC12 cells exposed to MPP+. The mitochondrial morphology was determined by laser confocal microscope. Western blot was used to measure the protein expression levels of mitochondrial fusion- and fission-related proteins, including OPA1, MFN2, DRP1 and Fis1. The results showed that pretreatment with different concentrations of Eda antagonized MPP+-induced PC12 cell damage in a dose-dependent manner. The PC12 cells treated with MPP+ showed mitochondrial fragmentation, up-regulated DRP1 and Fis1 protein expression levels, and down-regulated MFN2 and OPA1 protein expression levels. Eda could reverse the above changes in the MPP+-treated PC12 cells, but did not affect Fis1 protein expression. These results suggest that Eda has a protective effect on the mitochondrial fusion disruption induced by MPP+ in PC12 cells. The mechanism may be related to the up-regulation of OPA1/ MFN2 and down-regulation of DRP1.


Key words: edaravone; MPP+; PC12 cells; mitochondrial fission; mitochondrial fusion

收稿日期：2019-11-30　　录用日期：2020-02-13
通讯作者：宋成洁　　E-mail: songcj09@xzhmu.edu.cn
DOI: 10.13294/j.aps.2020.0023
引用本文：
焦阳, 郑月, 宋成洁. 依达拉奉对MPP⁺处理的PC12细胞线粒体融合和分裂平衡的保护作用[J]. 生理学报 2020; 72 (2): 249-254.
JIAO Yang, ZHENG Yue, SONG Cheng-Jie. Protective effect of edaravone on balance of mitochondrial fusion and fission in MPP⁺-treated PC12 cells. Acta Physiol Sin 2020; 72 (2): 249-254 (in Chinese with English abstract).



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