【摘要】 目的探讨2,2’,4,4’-四溴联苯醚(2,2’,4,4’-tetrabromodiphenyl ethers,PBDE-47)对人神经母细胞瘤SH-SY5Y细胞自噬水平的影响。方法取对数生长期的SH-SY5Y细胞,分别以终浓度为0(溶剂对照)、1、5、10μmol/L的PBDE-47染毒处理24 h后,采用透射电子显微镜观察细胞内双层膜自噬体结构;以自噬囊泡示踪剂单丹黄酰尸胺(MDC)染色检测自噬囊泡生成情况;采用Western blotting技术检测自噬相关蛋白LC3、Beclin1、P62的表达水平。结果电镜超微结构观察结果显示,对照组细胞内未见异常变化,1μmol/L染毒组细胞内可见轻微的线粒体扩张但未见明显的双层膜自噬体结构,5μmol/L染毒组细胞内可见明显的线粒体空泡及双层膜自噬体结构,10μmol/L染毒组细胞内观察到的双层膜自噬体结构最多;与对照组相比,5、10μmol/L PBDE-47染毒组MDC荧光强度升高,MDC阳性细胞比例增加;此外,5、10μmol/L PBDE-47染毒组自噬相关蛋白LC3、Beclin1、P62的表达水平与对照组相比均明显升高,且差异具有统计学意义(P<0.05)。结论 PBDE-47可诱导SH-SY5Y细胞自噬水平的增加。更多还原

【Abstract】 Objective To explore the effects of 2, 2’, 4, 4’-tetrabromodiphenyl ethers(PBDE-47) on autophagy level in human neuroblastoma SH-SY5Y cells. Methods Exponentially growing cells were incubated with PBDE-47 at the doses of 0(DMSO control), 1, 5 and 10 μmol/L respectively for 24 h in vitro. Ultrastructure of autophagosomes, monodansylcadaverine(MDC)-staining autophagic vesicles and the expression levels of autophagy relevant proteins of LC3, Beclin1 and P62 were determined by transmission, electron microscope, fluorescence microscopy and Western blotting. Results The results suggested that ultrastructure of SH-SY5Y cells were obviously changed by forming autophagosomes with double membrane, along with dilation and degeneration of mitochondria in the 5 and 10 μmol/L PBDE-47-treated groups compared with the control group, and 10 μmol/L PBDE-47-treated group cells could be observed within the most; 5 and 10 μmol/L PBDE-47-treated groups showed an increased relative fluorescence intensity of MDC, namely MDC-positive cell ratio increased. Meanwhile the expression levels of LC3, Beclin1, and P62 protein in 5, 10 μmol/L PBDE-47-treated groups were remarkably increased compared with controls(P<0.05). Conclusion PBDE-47 may increase the level of autophagy in SH-SY5Y.更多还原