PBDE-47对人神经母细胞瘤细胞凋亡和线粒体膜电位及细胞色素C蛋白表达的影响

Effect of PBDE-47 on apoptosis,mitochondrial membrane potential and protein expression of cytochrome C in SHSYSY cells

目的探讨PBDE-47对人神经母细胞瘤(SH-SY5Y)细胞线粒体膜电位和细胞色素C(Cyt C)蛋白表达的影响及其与细胞凋亡的关系。方法将处于对数生长期的SH-SY5Y细胞分别加入终浓度为0(溶剂对照组)、1、5、10μmol/L的PBDE-47暴露24 h。采用流式细胞术检测细胞凋亡率和线粒体膜电位变化,采用Western blot技术检测胞浆与线粒体细胞色素C蛋白表达的水平。结果与溶剂对照组相比,10μmol/L PBDE-47染毒组SH-SY5Y细胞凋亡率和5、10μmol/L PBDE-47染毒组胞浆内细胞色素C蛋白表达水平均明显升高,10μmol/L PBDE-47染毒组线粒体膜电位和各浓度PBDE-47染毒组线粒体内细胞色素C蛋白表达水平均明显下降,差异均有统计学意义(P<0.05)。且随着PBDE-47染毒浓度的升高,SH-SY5Y细胞凋亡率和胞浆内细胞色素C蛋白表达水平呈升高趋势,线粒体内细胞色素C蛋白表达水平呈下降趋势。结论 PBDE-47可通过引起线粒体膜通透性转运孔开放和细胞色素C的释放介导SH-SY5Y细胞凋亡,从而对SH-SY5Y细胞产生损伤。

Objective To explore the effects of PBDE-47 on mitochondrial membrane potential （AXiS＇m） and protein expression level of cytoehrome c （Cyt C） in human neuroblastoma cells （SH-SYSY cells）, and to study the relationship between them and apoptosis. Methods The SH-SYSY cells of logarithmic phase were exposed to final concentrations of 0 （solvent control group）, 1,5,10 μmol/L PBDE-47 for 24 h in vitro. The percentages of apoptosis and the changes of △ψm were analyzed by flow cytometry, and the protein expression levels of cytosolic and mitochondrial Cyt C were detected by Western blot. Results Compared with the solvent control group, the percentage of apoptosis increased significantly （P〈0.05） in 10 μmol/L PBDE-47 treated group,and eytosolic Cyt C protein expression levels increased significantly in 5 and 10 μmol/L PBDE-47 treated groups（P〈0.05）;△ψm decreased（P〈0.05） in 10 μmol/L PBDE-47 treated group,and mitochondrial Cyt C protein expressions in various concentrations of PBDE-47 decreased significantly（P〈0.05 ） compared with those in the solvent control group. As the concentrations of PBDE-47 increased, the apoptosis rates and protein expression level of cytosolic cytochrome C in SH-SYSY cell showed an increasing trend, and the protein expression level of mitochondrial eytochrome C revealed a declining trend. Conclusion PBDE-47 may induce SH-SYSY cells apoptosis by mitochondrial permeability transition pore opening and the release of Cyt C ,therefore it may cause the injury of SH-SYSY cells.