Effect of LPS and glutathione depletion on gene expression profiling in mouse macrophage-like cells

By microarray gene expression profiling, we investigated whether glutathione (GSH) has a signaling role in the response of RAW 264.7 cells to lipopolysaccharide (LPS). GSH was depleted by prior incubation with 120 µM buthionine sulfoximine (BSO) for 24 h, then cells were treated with 10 ng/ml LPS for 2 or 6 h. DNA microarray analysis was performed in triplicate samples in control and GSH-depleted cells, in the presence or absence of LPS. 4 conditions, 3 replicates, at 2 h and 6 h

本研究采用基因表达芯片谱分析（microarray gene expression profiling）技术，探究谷胱甘肽（glutathione, GSH）在RAW 264.7细胞对脂多糖（lipopolysaccharide, LPS）的应答过程中是否发挥信号调控作用。具体实验方案为：先将细胞与120 μM丁硫氨酸-亚砜亚胺（buthionine sulfoximine, BSO）共同孵育24小时以耗尽细胞内GSH，随后以10 ng/mL的LPS处理细胞2小时或6小时。本实验在对照组与GSH耗竭组细胞中，分别设置LPS处理与未处理的分组，每组均设置三份重复样本开展DNA芯片分析；实验共涵盖4种实验条件，覆盖2小时与6小时两个检测时间点。