论文部分内容阅读
目的探讨氟对人神经母细胞瘤SH-SY5Y细胞自噬水平的影响。方法用不同浓度(20、40、60 mg/L)的氟化钠(Na F)染毒SH-SY5Y细胞24 h,采用透射电子显微镜观察细胞内自噬体超微结构;吖啶橙染色观察细胞内自噬囊泡的生成;Western blotting技术检测自噬延伸相关蛋白Atg5和LC3-II与自噬降解相关蛋白P62的表达水平。结果透射电子显微镜下可观察到细胞内自噬体结构,且随NaF剂量的增加自噬体数量明显减少;吖啶橙染色可见,与对照组相比,40和60 mg/L NaF染毒组自噬囊泡数量逐渐减少,尤以60 mg/L NaF染毒组减少最为明显;Western blotting检测发现,与对照组相比,Atg5和LC3-II的表达水平呈剂量依赖性降低,而P62的表达水平呈剂量依赖性升高,且差异具有统计学意义(P<0.05)。结论氟可抑制SH-SY5Y细胞的自噬水平。
Objective To investigate the effect of fluoride on autophagy in human neuroblastoma SH-SY5Y cells. Methods SH-SY5Y cells were exposed to NaF at different concentrations (20, 40 and 60 mg / L) for 24 h. The ultrastructures of the autophagosomes were observed by transmission electron microscopy. Acridine orange staining The formation of autophagic vesicles was detected by Western blotting. The expressions of autophagy-related proteins Atg5 and LC3-II and autophagy-related protein P62 were detected by Western blotting. Results The structure of autophagosomes was observed under transmission electron microscope. The number of autophagosomes decreased with the increase of NaF dose. Acridine orange staining showed that compared with the control group, 40 and 60 mg / L NaF exposure groups The number of autophagic vesicles decreased gradually, especially in the 60 mg / L NaF group. Western blotting showed that the expression levels of Atg5 and LC3-II were decreased in a dose-dependent manner compared with the control group, while the expression of P62 The expression level increased in a dose-dependent manner, and the difference was statistically significant (P <0.05). Conclusion Fluoride can inhibit autophagy in SH-SY5Y cells.