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目的探讨促红细胞生成素(erythropoietin,EPO)对1-甲基-4-苯基吡啶离子(MPP+)诱导的PC12细胞变性损伤的保护作用及机制。方法用MPP+处理PC12细胞制作帕金森病细胞模型,采用四甲基偶氮唑蓝法检测暴露于不同浓度EPO后细胞的活性;流式细胞术与DNA断端原位标记法(terminal deoxynucleotidyl transferase dUTPnick end labeling, TUNEL)检测各组的细胞凋亡率;免疫印迹法检测不同处理组PC12细胞Bcl-2和Bax的表达,并采用荧光法观察不同处理组PC12细胞活性氧(reactive oxygen species,ROS)与线粒体膜电位水平以及caspase-3活性的变化。结果 MPP+可以使PC12细胞存活率下降,凋亡率增高;同时PC12细胞内ROS增多,线粒体膜电位下降。MPP+还可以明显地提高Bax/Bcl-2比值并激活caspase-3。而EPO可以抑制这些由MPP+引发的改变,并在1 U/mL时发挥最大保护作用。结论 EPO可抑制MPP+诱导的PC12细胞死亡,其作用机制可能与其自身抗氧化和抗凋亡的特性有关。
Objective To investigate the protective effect and mechanism of erythropoietin (EPO) on degeneration of PC12 cells induced by 1-methyl-4-phenylpyridinium ion (MPP +). Methods The PC12 cells were treated with MPP + to produce Parkinson’s disease model. The cell viability was detected by MTT method after exposed to different concentrations of EPO. Flow cytometry and terminal deoxynucleotidyl transferase (dUTPnick) The expression of Bcl-2 and Bax in PC12 cells of different treatment groups was detected by Western blotting. The expression of reactive oxygen species (ROS) in PC12 cells of different treatment groups was observed by fluorescence assay. And mitochondrial membrane potential levels and caspase-3 activity changes. Results MPP + could decrease the survival rate of PC12 cells and increase the apoptosis rate. At the same time, ROS increased and mitochondrial membrane potential decreased in PC12 cells. MPP + also significantly increased the Bax / Bcl-2 ratio and activated caspase-3. EPO, however, suppressed these MPP + -independent changes and exerted maximum protection at 1 U / mL. Conclusion EPO can inhibit MPP + -induced PC12 cell death, and its mechanism may be related to its own anti-oxidation and anti-apoptotic properties.