BDE47对Neuro-2a细胞毒性效应及作用机制

探讨2,2',4,4'-四溴联苯醚(2,2',4,4'-tetrabromodiphenylether,BDE 47)对神经细胞Neuro-2a的毒性影响及机制。将Neuro-2a细胞暴露于浓度为6.25、12.5、25、50、100μmol·L-1的BDE 47,采用MTT法检测细胞存活率、荧光探针DCFH-DA检测细胞活性氧生成量、吖啶橙检测溶酶体膜通透性、罗丹明123检测细胞线粒体膜电位、Annexin V-FITC检测细胞凋亡、Western blot检测组织蛋白酶B(Cathespin B)表达。结果显示,与对照组相比,12.5、25、50、100μmol·L~(-1)BDE 47显著降低Neuro-2a细胞存活率和细胞线粒体膜电位(P0.05);6.25、12.5、25、50、100μmol·L~(-1)BDE 47显著诱导活性氧含量升高(P0.05),增加Neuro-2a细胞溶酶体膜通透性(P0.05),诱导Neuro-2a细胞凋亡(P0.05),升高Cathespin B蛋白表达(P0.05)。结果表明,BDE 47可能通过介导溶酶体-活性氧-线粒体环路,诱导Neuro-2a细胞凋亡。

Toxic Effect and Mechanism of BDE 47 on Neuro-2a Cells

In order to explore the neurotoxicity of BDE 47, Neuro￣2a cells were exposed to 6.25, 12.5, 25, 50 and 100 μmol?L￣1 BDE 47 for 24 h. MTT was used to evaluate the cell viability, DCFH￣DA was used to detect the pro￣duction of reactive oxygen species (ROS), acridine orange (AO) was used to evaluate the lysosomal membrane per￣meability (LMP), rhodamine 123 was used to observe the mitochondrial membrane potential (MMP), Annexin V￣FITC was used to investigate the apoptosis, Western blot was used to detect the protein expression of cathepsin B. The results showed that compared with the control, 12.5, 25, 50, 100μmol?L￣1 BDE 47 decreased the viability and MMP of Neuro￣2a cells (P<0.05), respectively. And 6.25, 12.5, 25, 50, 100 μmol?L￣1 BDE 47 increased the pro￣duction of ROS, LMP, apoptosis and the expression of cathepsin B (P<0.05), respectively. These results indicated that BDE 47 might induce apoptosis of Neuro￣2a cells via lysosomal￣ROS￣mitochrondial cross￣talk pathway.