磁性三氧化二铁纳米粒子对小鼠腹腔巨噬细胞的氧化损伤作用(英文)  

Oxidative injury of magnetic ferric oxide nanoparticles to peritoneal macrophage in mice

作　　者：王晓娜[1] 唐萌[1] 张婷[1] 杨磊[1] 夏婷[1] 曾垂焕[1] 熊丽林[1] 张宇[2] 顾宁[2] 

机构地区：[1]东南大学公共卫生学院 [2]东南大学生物科学与医学工程系

出　　处：《中国组织工程研究与临床康复》2007年第13期2575-2577,2585,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家重点基础研究发展计划973项目资助(2006CB705602);国家自然科学基金项目资助(30671782)~~

摘　　要：背景:已有报道表明纳米级的Fe2O3产生的细胞毒性与细胞的脂质过氧化存在一定的联系。氧化铁纳米粒子是否对巨噬细胞产生毒性,其毒性机制与氧化作用有何关联?目的:观察不同浓度Fe2O3纳米粒子对巨噬细胞的氧化损伤作用。设计:观察对比实验。单位:东南大学公共卫生学院。材料:RAW264.7细胞为小鼠腹腔巨噬细胞,购自中国科学院上海细胞所。Fe2O3纳米粒子(30nm)悬浮液由东南大学生物医学工程系制备提供。实验前先进行预处理:将Fe2O3纳米粒子悬浮液置60℃水浴中10h,然后37℃水浴过夜。如此反复3次,灭菌处理。小瓶分装,4℃保存。DMEM高糖培养液(美国Gibco公司);胰蛋白酶(美国Difco公司,进口分装);新生牛血清(杭州四季青公司);过氧化氢、羟自由基、超氧阴离子自由基、乳酸脱氢酶、超微量ATP酶和考马斯亮蓝蛋白含量测定试剂盒(南京建成生物技术公司)。方法:实验于2006-03/07在东南大学公共卫生学院劳动卫生与环境卫生学系实验室完成。RAW264.7细胞用DMEM培养基于37℃、体积分数为0.05的CO2培养箱中进行培养。每日用倒置显微镜观察细胞生长情况,取生长状态良好的对数生长期细胞进行试验。①细胞中氧自由基的检测:将密度为1.5×105L-1的巨噬细胞接种于24孔培养板,每孔1mL,37℃、体积分数为0.05的CO2培养箱中培养24h后,以质量浓度为1.0700、0.5350和0.2675g/L的Fe2O3纳米粒子(30nm)悬浮液染毒细胞为纳米粒子干预组,并设生理盐水为溶剂对照组,24h后终止培养,染毒结束后,低温条件下用玻璃匀浆器破碎细胞,按试剂盒说明,分别测定细胞中过氧化氢、羟自由基、超氧阴离子自由基。②培养液中乳酸脱氢酶活性及膜ATP酶活性的测定:纳米粒子干预组及对照组干预方法同上,染毒结束后,检测培养液中乳酸脱氢酶活性及膜ATP酶活性,乳酸脱氢酶活性的检测采用南京建成生物工程研BACKGROUND ： Reports have demonstrated that cytotoxicity produced by ferric oxide （Fe2O3） nanoparticles is associated with cellular lipid peroxidation. Whether Fe2O3 nanoparticles have toxicity to macrophages, and what is the association of toxic mechanism and oxidization？ OB3ECTIVE： To observe the effects of different concentrations of Fe2O3 nanoparticles on the oxidative damage of macrophages. DESIGN ： A controlled observation experiment SEI-rING： School of Public Health, Southeast University MATERIALS： RAW264.7 cells were peritoneal macrophages of mouse and purchased from Shanghai Institute of calls, Chinese Academy of Sciences. Fe2O3 nanoparticles （30 nm） suspension was provided by Department of Biomedical Engineering, Southeast University）. Fe2O3 nanoparticle suspension was placed in 60 ℃ water for 10 hours, then in 37 ℃ water overnight. This procedure was repeated 3 times for germicidal treatment. Then, the suspension was packed into small bottles and stored at 4 ℃ for later use. DMEM high glucose culture fluid （Gibco Company, USA）; trypsinase （Difco Company, USA, imported）; new-born calf serum（Sijiqing Company, Hangzhou）; hydrogen dioxide （H2O2, Gibco Company）; Kits for measuring hydrogen dioxide（H2O2）, hydroxy radical （-OH）, superoxide anion radical （O2·^-）, lactic acid dehydrogenase, ultramicro ATP enzyme and Coomassie brilliant blue protein levels （Jiancheng Biotechnique Co., Ltd., Nanjing）, METHODS ： This experiment was carried out in the laboratory of Department of Labor and Environmental Health, School of Public Health, Dongnan University between March 2006 and July 2006. RAW264.7 cells （Abelson murine leukemia virus-induced tumor） were cultured in DMEM （Gibco Company） containing 100 g/L fetal bovine serum, 100 000 U/L penicillin and 100 mg/L streptomycin in the environment of 5% CO2. Cell growth was observed under an inverted microscope. Cells, which were at good exponential phase of growth, were chosen for test. ①Detection of

关 键 词：FE2O3 纳米粒子 巨噬细胞 氧化应激 毒性 

分 类 号：R349.5[医药卫生—基础医学]