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研究丙戊酸钠(sodium valproate,VPA)对抗鱼藤酮(Rotenone)诱导的SH-SY5Y细胞损伤的作用及线粒体机制。以1,10μmol/L VPA预处理SH-SY5Y细胞3 h,再加入400 nmol/L Rotenone作用24 h。MTT法检测与相差显微镜观察相结合,分析VPA对抗Rotenone损伤的作用;JC-1染色法与Mito-Tracker染色法分析线粒体膜电位及线粒体数量的变化;Clark氧电极法检测细胞呼吸功能;DCFH-DA探针法检测细胞中ROS的含量;并在离体线粒体上观察VPA对Ca2+诱导的线粒体肿胀的影响。结果发现,1,10μmol/L VPA预处理SH-SY5Y细胞3 h可对抗400 nmol/L Rotenone引起的细胞损伤,并且可以提高损伤细胞中线粒体的膜电位,增加线粒体的数量,此外,还可以增强损伤细胞的呼吸功能,降低细胞中ROS的含量,但VPA并不能直接作用于离体的线粒体发挥神经保护作用。由此,VPA具有良好的神经保护作用,其机制与增强线粒体功能和数量、从而改善细胞功能有关,这为其应用于帕金森病的预防与治疗提供了实验依据。
To investigate the effects of sodium valproate (VPA) on rotenone-induced injury of SH-SY5Y cells and its mitochondrial mechanism. SH-SY5Y cells were pre-treated with 1,10μmol / L VPA for 3 h, then treated with 400 nmol / L Rotenone for 24 h. MTT assay and phase contrast microscopy were used to analyze the effect of VPA on Rotenone injury. The changes of mitochondrial membrane potential and mitochondria were analyzed by JC-1 staining and Mito-Tracker staining. The respiratory function of DCFH- DA probe method was used to detect the content of ROS in cells. The effect of VPA on Ca2 +-induced mitochondrial swelling was observed in isolated mitochondria. The results showed that 1, 10μmol / L VPA pretreatment SH-SY5Y cells 3 hours against 400 nmol / L Rotenone-induced cell damage, and can increase the mitochondrial membrane potential damage cells, increasing the number of mitochondria, in addition, can also be enhanced Damaging the respiratory function of cells and reducing the content of ROS in cells. However, VPA can not directly exert neuroprotective effect on isolated mitochondria. Thus, VPA has a good neuroprotective effect, and its mechanism is related to the enhancement of mitochondrial function and quantity to improve cell function, which provides an experimental basis for the prevention and treatment of Parkinson’s disease.