不同表面修饰的氧化铁纳米颗粒对A549细胞的毒性及DNA损伤  

Induction of cytotoxicity and DNA damage by iron oxide nanoparticles with different surface modifications in A549 cells

作　　者：文海若 郭雅娟 黄芝瑛[2] 王雪[1] 淡墨[1] WEN Hairuo;GUO Yajuan;HUANG Zhiying;WANG Xue;DAN Mo(National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Key Laboratory of Beijing for Nonclinical Safety Evaluation Research of Drugs,Beijing 100176;School of Phamaceutical Sciences,Sun Yat-sen University,Guangzhou 510006,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]中山大学药学院,广东广州510006

出　　处：《癌变．畸变．突变》2018年第6期413-421,共9页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(81401517)

摘　　要：目的:比较不同表面修饰的氧化铁纳米颗粒(IONP_s)对A549细胞的细胞毒性、染色体和DNA损伤及其作用机制的差异。方法:比较相同粒径范围(约5nm)的胺基表面修饰的纳米氧化铁颗粒(Amine-IONP)和聚乙二醇表面修饰的纳米氧化铁颗粒(PEG-IONP)对A549细胞存活率和细胞周期的影响;使用高内涵法检测细胞经IONP_s处理后的胞内活性氧簇(ROS)含量、线粒体膜电位(MMP)和内质网(ER)状态的变化;采用体外胞质分裂法微核试验和彗星电泳评价IONP_s对染色体和DNA完整性的影响。结果:两种纳米氧化铁颗粒处理48h对A549细胞生长抑制率均小于20%。相同浓度条件下,PEG-IONP主要表现为对A549细胞G0/G1期阻滞,自20μg/mL起即明显减少S期细胞比率(P<0.01),320μg/mLPEG-IONP处理24h后可诱导p21与p53表达水平显著升高(P<0.05)。给药48h时,Amine-IONP作用后细胞ROS、MMP及ER水平显著性改变的起始浓度分别为20、20和80μg/mL,而PEG-IONP作用后产生显著性改变的起始浓度分别为40、40和160μg/mL。此外,与PEG-IONP比较,Amine-IONP可在较低浓度条件下诱导微核和彗星拖尾形成(Amine-IONP的起始浓度为20和80μg/mL;而PEG-IONP则为40和160μg/mL)。经氧自由基清除剂乙酰半胱氨酸和叔丁基对羟基茴香醚预处理后,两者导致的胞内ROS含量和尾部DNA百分率均明显降低(P<0.05)。结论:带正电荷的Amine-IONP更易于诱导A549细胞氧化应激及与之有关的DNA损伤;相比之下,PEG-IONP的细胞毒性和遗传毒性较弱,但除氧化损伤外也可通过抑制细胞周期干扰细胞增殖,作为肿瘤诊断试剂具有一定优势。OBJECTIVE:To compare the induction of cytotoxicity,chromosome aberrations and DNA damage among iron oxide nanoparticles(IONPs)with different surface-modifications in A549 cells.METHODS:The effects of amine-modified(Amine-IONP)and polyethylene glycol-modified iron oxide nanoparticles(PEG-IONP)of the same particle size range(5 nm)on cell viability and cell cycle were compared in A549 cells;the content of intracellular reactive oxygen species(ROS),mitochondrial membrane potential(MMP)and endoplasmi reticulum stress(ERS)in cells treated with IONPs were analyzed using high content screening method;the in vitro cytokinesis micronucleus test and Comet assay were performed to evaluate their effects on chromosome and DNA integrity.RESULTS:The growth inhibitory rates of both IONPs to A549 cells were less than 20%after 48 h.Under the same concentration,PEG-IONP exhibited a more significant effect on the G0/G1 cell cycle arrest(P<0.05):the initial concentration to significantly reduce the cell rate of S phase was 20μg/mL,and the expression of p21 and p53 was up-regulated by a treatment of 320μg/mL PEG-IONP for 24 h(P<0.05).After a 48 h treatment,the initial concentrations to show significant effects of Amine-IONP on ROS,MMP and ER were 20,20 and 80μg/mL,while for PEG-IONP they were 40,40 and 160μg/mL respectively.In addition,compared to PEG-IONP,Amine-IONP was able to induce the formation of micronuclei and Comet tail(the initial concentrations for Amine-IONP were 20 and 80μg/mL;while PEG-IONP were 40 and 160μg/mL respectively).After the pre-treatment with oxygen free radical scavengers N-acetylcysteine and butylated hydroxyanisole,both IONPs induced ROS and Comet DNA Tail were significantly inhibited(P<0.05).CONCLUSION:Positively charged Amine-IONP was more effective in inducing oxidative stress and DNA damage in A549 cells;whereas,PEG-IONP with relatively weaker cytotoxicity and genotoxicity interfered with cell proliferation,and showed advantages in the development of tumor diagnostic reagents.

关 键 词：氧化铁纳米颗粒 A549细胞 微核试验 彗星试验 氧化应激 细胞增殖 

分 类 号：R917[医药卫生—药物分析学]