小鼠腹腔巨噬细胞炎症模型的建立  

Inflammatory model of mouse peritoneal macrophages

作　　者：余功旺 黄文浩[1] 刘爱梅[1] 孙艳[1] 

机构地区：[1]广东药学院生命科学与生物制药学院,广东广州510006

出　　处：《广东药学院学报》2014年第6期766-770,共5页Academic Journal of Guangdong College of Pharmacy

基　　金：国家自然科学基金项目(81000923);广东省医学科学技术研究基金项目(B2009117);广东高校优秀青年创新人才培育项目(2009400)

摘　　要：目的体外建立小鼠腹腔巨噬细胞炎症模型,为体外研究中药抗炎机制提供实验模型。方法分离、培养小鼠腹腔巨噬细胞;形态学观察巨噬细胞的生长;流式细胞术鉴定所分离细胞的纯度;以不同质量浓度(0、0.1、1、10μg/m L)的脂多糖(LPS)作用细胞24 h及1μg/m L的LPS作用细胞不同时间(0、4、8、16、24、48 h),荧光定量PCR法检测细胞中IL-1β、IL-6和TNF-αmRNA的表达。结果获得的贴壁细胞具备巨噬细胞的形态特征,纯度为84.55%;在一定范围内,LPS以浓度依赖和时间依赖的方式上调IL-1β、IL-6和TNF-αmRNA的表达。结论 1μg/m L的LPS作用细胞24 h能建立较为合理的炎症模型。Objective To establish lipopolysaccharide（ LPS）-induced inflammatory model of mouse peritoneal macrophages in vitro for evaluation of the anti-inflammatory mechanism of Chinese medicine. Methods The peritoneal macrophages from mice were isolated and purified. The purity of macrophages was identified by flow cytometry. Then macrophages were stimulated with 0.1,1 and 10 μg / m L LPS for 24 h or treated with 1 μg / m L LPS for different times（ 0,4,8,16,24 and 48 hours）. The expression of IL-1β,IL-6 and TNF-α mRNA was detected by real-time quantitative-PCR. Results The harvested adherent cells showed the morphological characteristics of macrophages and the purity was 84.55%. In a certain range,LPS enhanced the expression of IL-1β,IL-6 and TNF-α mRNA in a time-dependent and concentration-dependent manner. Conclusion The inflammatory model will be established after macrophages stimulation with 1 μg / m L LPS for 24 h.

关 键 词：巨噬细胞 脂多糖 炎症模型 

分 类 号：R-332[医药卫生]