| 宋蕊,刘晨旭,张睿,等.1-甲基-4-苯基吡啶离子诱导大鼠肾上腺嗜铬细胞瘤PC12细胞发生铁死亡的实验观察[J].安徽医药,2017,21(2):220-225. |
| 1-甲基-4-苯基吡啶离子诱导大鼠肾上腺嗜铬细胞瘤PC12细胞发生铁死亡的实验观察 |
| Investigation of MPP+induced ferroptosis in PC12 cells |
| 投稿时间：2016-08-21 |
| DOI： |
| 中文关键词: 帕金森病 铁死亡 1-甲基-4-苯基吡啶离子 铁死亡抑制剂 PC12细胞 |
| 英文关键词: Parkinson's disease Ferroptosis MPP+ Ferrostatin-1 PC12 cells |
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| 中文摘要: |
| 目的 进行1-甲基-4-苯基吡啶离子(1-methyl-4-phenyl pyridine,MPP+)诱导大鼠肾上腺嗜铬细胞瘤PC12细胞铁死亡(Ferroptosis)的研究。方法 采用噻唑蓝法(MTT)检测细胞存活率和筛选铁死亡抑制剂(Ferrostatin-1)的最佳作用浓度；倒置显微镜观察Ferrostatin-1对PC12细胞的保护作用；MDC染色检测细胞自噬；ELISA法检测caspase-3的酶活性；流式细胞术检测活性氧ROS。结果 1 mmol·L-1 MPP+对PC12细胞有明显抑制作用；5 μmol·L-1 Ferrostatin-1能够明显提高PC12细胞的存活率；倒置显微镜观察结果表明Ferrostatin-1预保护后,PC12细胞损伤明显减少；MDC染色检测细胞自噬和ELISA法检测caspase-3的酶活性结果显示Ferrostatin-1对MPP+诱导的PC12细胞自噬和凋亡的保护作用不明显；ROS活性氧检测结果显示ROS在胞内增加,可能发生氧化应激,加Ferrostatin-1后ROS荧光强度减弱。结论 Ferroptosis能够显著抑制MPP+诱导的PC12细胞损伤,且Ferrostatin-1对细胞的自噬和凋亡作用不明显,推测出MPP+诱导PC12细胞的损伤可能有Ferroptosis的存在。 |
| 英文摘要: |
| Objective To study MPP+ induced ferroptosis in PC12 cells.Methods The cell viability was detected and the optimum concentrations of Ferrostatin-1 were screened by MTT assay;the protective effect of Ferrostatin-1 on PC12 cells were by inverted microscope observation,the cell autophagy were inspected by MDC staining;caspase-3 enzyme activity were tested by ELISA method,the intracellular ROS was analyzed by ROS assay kit.Resluts The results manifested that 1 mmol·L-1 MPP+ had obvious inhibitory effect on PC12 cells.5 μmmol·L-1 Ferrostatin-1 can obviously improve the survival rate of PC12 cells.After Ferrostatin-1 incubation,the injury on PC12 cells were significantly reduced.MDC staining exhibited that the protection effect of Ferrostatin-1 on the autophagy of PC12 cells induced by MPP+ was not obvious.Caspase-3 activity tests showed that the protection effect of Ferrostatin-1 on the apoptosis of PC12 cells induced by MPP+ was not remarkable.The intracellular ROS was increased significantly,which lead to oxidative stress.And incubation with Ferrostatin-1 can reduce the intensity of fluorescence.Conclusion Ferrostatin-1 can significantly inhibit PC12 cells′ injury induced by MPP+,and the effect of Ferrostatin-1 on autophagy and apoptosis is not significant.It assumed the existence of ferroptosis on the model of MPP+ damaged PC12 cells. |
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