Genome-wide anyalysis of gene expression during prolonged endotoxin tolerance in Aoah -/- mice. Mus musculus

Analysis of gene expression in explanted peritoneal macrophages from Aoah -/- and Aoah +/+ mice treated with LPS 21 days prior to harvest. Explanted peritoneal machrophages were challenged with LPS or control (PBS). The study seeks to characterize global gene expression in the state of prolonged LPS tolerance induced in mice lacking the LPS-inactivating enzyme Aoah. Overall design: Groups of 9 C57BL/6 Aoah+/+ or Aoah-/- mice were given i.p. injections of 10 µg E. coli LPS/mouse. Twenty-one days later (when Aoah-/- mice remain tolerant and Aoah+/+ mice have recovered), peritoneal macrophages were harvested and the yields from three mice were pooled to form 3 samples per group (i.e., three samples of Aoah+/+ and Aoah-/- mice, with each sample comprised of peritoneal macrophages from 3 mice). Next day, cells were challenged with LPS or PBS and whole RNA was isolated 2 hours later and used for microarray experiments.

本研究针对取材前21天接受脂多糖（LPS）处理的Aoah+/+与Aoah-/-小鼠的离体腹腔巨噬细胞开展基因表达分析。将分离得到的腹腔巨噬细胞分别用LPS或对照试剂磷酸盐缓冲液（PBS）进行刺激。本研究旨在表征缺乏脂多糖灭活酶Aoah的小鼠体内诱导的长期LPS耐受状态下的全基因表达特征。实验设计概述：将9只一组的C57BL/6背景Aoah+/+与Aoah-/-小鼠，每只腹腔注射10μg大肠杆菌LPS。21天后（此时Aoah-/-小鼠仍处于耐受状态，而Aoah+/+小鼠已恢复），收集腹腔巨噬细胞，将每3只小鼠的细胞混合为1份样本，每组共制备3份样本（即Aoah+/+与Aoah-/-组各3份样本，每份样本均来自3只小鼠的腹腔巨噬细胞）。次日，将细胞分别用LPS或PBS刺激，2小时后提取总RNA，用于微阵列（microarray）实验。