纳米氧化锌致细胞凋亡的机制研究  

Mechanism of nano-zinc oxide-induced apoptosis

作　　者：杨通旺 张博闻[1] 吴涛[1] 吴刚[1] 

机构地区：[1]包头医学院研究生学院,内蒙古包头014040

出　　处：《工业卫生与职业病》2016年第3期190-193,共4页Industrial Health and Occupational Diseases

基　　金：国家自然基金(81160341)

摘　　要：目的研究氧化应激在纳米氧化锌引起的人非小细胞肺癌细胞(H1299)细胞自噬和凋亡之间的作用。方法实验分为实验组和对照组,对照组给予正常的培养基而实验组在培养基中分别给予2.5、10和25μg/ml(低,中,高剂量组)浓度的纳米氧化锌。向细胞中转染绿色荧光蛋白-微管相关蛋白1轻链3(GFP-LC3)质粒,通过观察绿色荧光斑点确定自噬的发生。Western blotting检测LC3-Ⅱ来验证绿色荧光斑点确定的细胞自噬。钙黄绿素乙酰甲酯(calcein AM)和碘化丙啶(PI)染色来确定活细胞和晚期凋亡细胞。超氧化物阴离子荧光探针(DHE)染色检测细胞内活性氧(ROS)的聚集。应用动态光散射测定纳米氧化锌在培养基中的分散状态。结果对照组细胞有少量的DHE被氧化成红色,也出现了绿色荧光斑点的聚集,但没有细胞被PI染成红色。低剂量组被氧化的DHE开始增多,绿色荧光斑点开始增多,少量细胞被PI染成红色;中、高剂量组中大量DHE被氧化,绿色荧光斑点显著增多,大量细胞被PI染成红色。实验组纳米氧化锌在培养基中分散良好。结论纳米氧化锌在H1299细胞中引起了氧化应激,氧化应激诱发了细胞的自噬,细胞自噬功能的紊乱诱导了细胞的凋亡。Objective To explore the rule of oxidative stress which introduced by nano-zinc oxide between autophagy and apoptosis in non-small cell lung cancer（H1299）cells line.Methods The study were divided into experimental and control groups.The control group was given normal medium,and the experimental groups were given different dose of nano-zinc oxide（2.5,10 and 25μg/ml）.The GFP-LC3 plasmid were translated into the cells,green fluorescent spots were considered autophagosome.LC3- Ⅱwhich is the biomarker of the autohagosome was detected by Western blotting.The living and late apoptosis cells were detected by Calcein AM and PI stain,respectively.The accumulation of intracellular reactive oxygen species（ROS）was detected by DHE.The dispersed state of the nano-zinc oxide was detected by dynamic light scattering in the medium.Results There were little cells stained to red by the DHE and accumulated with green fluorescent spots,but the cells were not stained to red by PI.The cells which stained to red by the DHE increased,green fluorescent spots were accumulated in the low-dose group,and there were some cells stained to red by PI.The majority of the cells were stained to red by the DHE,green fluorescent spots were accumulated in both the medium and high dose groups,and the cells were stained to red by the PI at the same time.The nano-zinc oxide was dispersed well in the medium.Conclusions The disorder of the autophagy that caused by the oxidative stress which induced by the nano-zinc oxide lead apoptosis in the H1299 cells line.

关 键 词：靶向运输 纳米粒子 氧化应激 凋亡 自噬 

分 类 号：R99[医药卫生—毒理学]