甲酸钠诱导视网膜光感受器细胞自噬

Sodium formate induces retinal photoreceptor cells autophagy

目的近些年来,发生过很多因饮用假酒致甲醇中毒的事件。本研究旨在探讨甲酸钠诱导视网膜光感受器细胞(661W细胞)损伤过程中对细胞自噬水平的影响。方法将15和30mM浓度的甲酸钠分别加入661W细胞培养基作用24h后,用MDC荧光染色来检测细胞自噬水平改变,透射电镜观察661W细胞自噬体的形成,Western blotting法检测LC3、p-mTOR、Beclin 1及JNK、p-JNK蛋白的表达,流式细胞仪分析检测细胞内活性氧的变化。结果与对照组比较,15或30mM甲酸钠处理661W细胞后,荧光显微镜下观察到细胞自噬水平增强,透射电镜下观察到自噬体形成,Western blotting法显示LC3-II,Beclin 1以及pJNK的蛋白表达水平上调,而p-mTOR的蛋白表达水平减少,流式细胞仪检测法显示甲酸钠处理的661W细胞内活性氧水平增加。结论甲酸钠可诱导661W细胞自噬,而自噬可能参与假酒中毒所致的视觉损伤过程。

Objective In recent years,alcoholic poisoning has occurred due to drinking fake wine. The aim of this study was to investigate the variation of cell autophagy level,in the process of sodium formate inducing retinal photoreceptor cells （661 w） damaged. Methods 661W cells were exposed to 15 and 30 mM concentration of sodium formate for 24h respectively, then, the autophagy level was detected by monodansylcadaverine staining. The autophagy formation of 661W cells was observed by transmission electron microscopy. The expression of rnierotubule associated protein LC3,p-mTOR, Beclin 1, JNK and p-JNK were evaluated by western blotting. Intracellular reactive oxygen species （ROS） were detected using 2＇,7＇. dichlorofluorescein diacetate （DCFH. DA） staining, followed by flow eytometric analysis. Results Compared with the control group,sodium formate at concentrations of 15 or 30 mM markedly increased the level of autophagy level was observed under fluorescence microscope. Autophagosomes were observed under transmission electron microscope. Western blotting showed that the levels of LC3-II. Beclin 1, p JNK exhibited increased. while the expression of p-mTOR protein was decreased. And flow cytometry showed that the level of reactive oxygen species increased. Conclusion Sodium formate can induce autophagy of 661W cells. Autophagy may be involved in the process of visual impairment caused by fakes poisoning.