甲酸钠诱导视网膜光感受器细胞凋亡

Sodium formate induces photoreceptor cells apoptosis

目的甲醇具有选择性神经毒性作用,世界各地因饮用假酒所致甲醇中毒事件时有发生,甚至有人因此而失明。我们将研究甲酸钠对视网膜光感受器细胞(661W细胞)的作用。方法将不同浓度的甲酸钠加入培养基的661W细胞中作用6-24h,通过MTT法检测各组细胞相对存活率,然后,将661W细胞暴露于15mM和30mM浓度的甲酸钠中24h,通过Hoechst 33342/PI染色法在荧光显微镜下观察细胞凋亡,用流式细胞仪检测细胞凋亡,用Western blotting法检测Bax,bcl-2,cleaved-caspase-3,cleaved-caspase-9的蛋白活性。结果 MTT结果显示:甲酸钠可以诱导661w细胞浓度和时间依赖性生存率下降;细胞染色、流式细胞仪检测结果显示:甲酸钠可以诱导661W细胞发生凋亡;Western blotting结果显示:15 mM和30 mM的甲酸钠作用661W细胞24h后,细胞Bax,cleavedcaspase-3,cleaved-caspase-9蛋白表达水平增加,而bcl-2蛋白表达水平降低。结论甲酸钠可以诱导光感受器细胞发生浓度和时间依赖性生存率下降,并且诱导661w细胞发生caspase依赖性细胞凋亡。

Objective Methanol intoxicated accidents caused by drinking fake wine take place now and then in the world. Some people become blindness due to methanol selective neurotoxicity. This study was to investigate the effects of sodium formate exposure on photoreceptor cells （661W cells）. Methods 661W cells were exposed to dfferent con- centrations of sodium formate for 6-24 h. The cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphen- yl-2H-tetrazolium bromide （MTT） assays. Then,661W cells were exposed to 15mM and 30 mM sodium formate for 24 h. Apoptosis was determined by Hoechst 33342/propidium iodide （PI） staining,visualized under a fluorescence micro- scope,and annexin V-FITC staining using flow cytometry. The activation of Bax,Bcl-2, cleaved-caspase-3 and cleaved- caspase-9,was evaluated by western blotting. Results MTT assay showed that, sodium formate treatment induced a decrease in 661W cell viability in a time- and dose-dependent manner;DCFH-DA staining and flow cytometry showed that , sdium formate induces 661W cells apoptosis. Western blotting showed that, 66 l W cells exposed to sodium formate for 24 h increased levels of Bax, eleaved-caspase-3 and cleaved-caspase-9, but decreased levels of Bcl-2. Conclusion These results suggest that sodium formate treatment reduced the cell viability in a time- and dose-dependent manner and triggers caspase-dependent apoptosis of 661W cells.