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

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500μmol/L1-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。

Protective effect of edaravone on balance of mitochondrial fusion and fission in MPP^+-treated PC12 cells

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.