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

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. Overall design: 4 conditions, 3 replicates, at 2 h and 6 h

本研究通过基因芯片（microarray）基因表达谱分析，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。实验先将细胞与120 μM 丁硫氨酸-亚砜亚胺（buthionine sulfoximine, BSO）孵育24小时以耗尽胞内GSH，随后使用10 ng/mL的LPS分别处理细胞2小时与6小时。针对对照组与GSH耗尽组细胞，分别设置有LPS处理与无LPS处理的分组，每组均设置3次生物学重复，开展DNA基因芯片（DNA microarray）分析。实验整体设计：共包含4种处理条件，每个条件设置3次重复，分别在2小时与6小时两个时间点完成采样与分析。