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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 investigate the effect of PBDE-47 on mitochondrial membrane potential and cytochrome C (Cyt C) protein expression in human neuroblastoma (SH-SY5Y) cells and its relationship with apoptosis. Methods SH-SY5Y cells in logarithmic growth phase were added to a final concentration of 0 (solvent control group), PBDE-47 at 1,5 and 10 μmol / L for 24 h. The apoptosis rate and mitochondrial membrane potential were detected by flow cytometry. The levels of cytochrome C protein in cytoplasm and mitochondria were detected by Western blot. Results Compared with the solvent control group, the apoptotic rate of SH-SY5Y cells in 10μmol / L PBDE-47 group and the intracytoplasmic cytochrome C protein expression in 5,10μmol / L PBDE-47 group were significantly increased, The mitochondrial membrane potential of mitochondrial membrane potential of PBDE-47 treated with 10μmol / L PBDE-47 and the concentration of cytochrome C protein of PBDE-47 treated group were significantly decreased (P <0.05). With the increase of PBDE-47 concentration, the apoptotic rate of SH-SY5Y cells and the level of intracellular cytochrome C protein expression increased, and the expression level of cytochrome C protein in mitochondria decreased. Conclusion PBDE-47 can induce the apoptosis of SH-SY5Y cells by causing the opening of mitochondrial permeability transmembrane opening and the release of cytochrome C, resulting in the injury of SH-SY5Y cells.