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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。
Objective To investigate the effects of 2,2 ’, 4,4’-tetrabromodiphenyl ethers (PBDE-47) on autophagy in human neuroblastoma SH-SY5Y cells . Methods SH-SY5Y cells in logarithmic growth phase were treated with PBDE-47 at a concentration of 0 (solvent control) and exposed to 1,5,10 μmol / L of PBDE-47 for 24 h. Transmission electron microscopy was used to observe the intracellular bilayers Membrane autophagosomes; autophagic vesicle formation was detected by autosomal vesicle tracer mono-dansyl-cysteic acid (MDC) staining; Western blotting was used to detect the expression of autophagy-related proteins LC3, Beclin1 and P62. Results The ultrastructural observation of electron microscopy showed that there was no abnormal change in the control cells. In the 1 μmol / L group, slight mitochondrial dilation was found but no obvious bilayer membrane autophagosome was observed. In the 5 μmol / L group The obvious mitochondrial vacuoles and bilayer membrane autophagosomes were observed in the cells. The highest bifurcation membrane autophagosome was observed in the cells treated with 10μmol / L. Compared with the control group, 5,10μmol / L PBDE- The fluorescence intensity of MDC increased and the percentage of MDC positive cells increased in PBMC-47-treated group. In addition, the expressions of autophagy-related proteins LC3, Beclin1 and P62 in PBDE-47 treated group were significantly higher than those in control group , And the difference was statistically significant (P <0.05). Conclusion PBDE-47 can induce the increase of autophagy in SH-SY5Y cells.