巨噬细胞缺氧复氧模型的制作方法及其生物学意义  

Establishment of a Hypoxia/Reoxygenation Model of Macrophage and Its Biological Significance

作　　者：张艳美[1] 廖晗[1] 郑付春[2] 李伟秋[3] 郑燕珊[1] 徐涵[1] 刘幸平[1] 石刚刚[1] 

机构地区：[1]汕头大学医学院药理学教研室,广东汕头515041 [2]汕头大学医学院第一附属医院药剂科,广东汕头515041 [3]汕头大学医学院分析细胞学实验室,广东汕头515041

出　　处：《汕头大学医学院学报》2013年第1期13-15,F0003,共4页Journal of Shantou University Medical College

基　　金：国家自然科学基金-广东省自然科学基金联合资助项目(U0932005);国家自然科学基金资助项目(81072633)

摘　　要：目的:建立一种简单可行的巨噬细胞缺氧复氧(H/R)模型。方法:采用制备缺氧液与氮气饱和相结合的方法制造细胞缺氧环境。巨噬细胞系RAW264.7分为对照组、缺氧(30、60和120min)复氧(15或30min)组及H/R+不同剂量的依达拉奉(EDA)组,借助于倒置荧光显微镜及流式细胞仪观测巨噬细胞内活性氧(ROS)浓度的变化,作为评判H/R模型成功与否的标准。结果:与各自对照组比,缺氧60min复氧15 min组即可引起ROS浓度明显升高(P<0.05),随着H/R时间延长,ROS生成呈进行性增加趋势。ROS清除剂EDA能剂量-依赖地抑制H/R所致ROS的大量生成。结论:应用制备缺氧液与氮气饱和相结合的方法可成功建立巨噬细胞H/R模型,该模型适于不同脏器缺血再灌注损伤共同分子机制的探讨及寻找共同治疗策略的相关研究。Objective： To establish a simple and feasible method of macrophage hypoxia/reoxygenation（H/R）.Metho-ds： The hypoxic condition was set up by a method of preparing hypoxia solution combining saturating with nitrogen gas.RAW264.7 macrophage cells were divided into control,H/R（30～120 minutes hypoxia followed by 15～30 minutes reoxygenation）and H/R＋edaravones（EDA）in different dosages groups.The levels of reactive oxygenspecies（ROS）,an index to determine of success for macrophage H/R model,were detected by inversion fluorescence microscope and flow cytometry.Results： Compared with the respective control groups,the levels of ROS in H∶60 minutes/R∶15 minutes group were significantly increased（P0.05）.During the study period,it significantly progressively increased with time.EDA,a scavenger of ROS,ameliorated H/R-induced increase of ROS level in a dose-dependent manner.Conclusion： The H/R model of macrophage can be made by preparing hypoxia solution combining saturating with nitrogen gas,which is applicable to studies for searching common molecular mechanism and treatment strategy of different organs I/R injury.

关 键 词：巨噬细胞 缺氧复氧 模型 氧化应激 

分 类 号：R363[医药卫生—病理学]