Liver gene expression profiles of Tribbles-homolog1 (Trib1)/Ldl-receptor (Ldlr) double-knockout mice against Ldlr-knockout mice

Genetic variants at the TRIB1 (Tribbles-homolog 1) gene locus are strongly associated with plasma lipid traits and the risk of coronary artery disease in humans. In this study, we analyzed the consequences of Trib1-deficiency (Trib1-/-) on hepatic gene expression in mice on the atherosclerosis-susceptible LDL-receptor deficient (Ldlr-/-) background. Therefore, Trib1-/- mice were crossed onto the Ldlr-/- background to generate double knockout mice (Trib1-/-Ldlr-/-) and fed a semisynthetic, modified AIN76 diet (0.02% cholesterol, 4.3% fat). At 20 weeks of age, Trib1-/-Ldlr-/- mice and Trib1+/+Ldlr-/- control mice were sacrificed and liver tissue was snap frozen in liquid nitrogen and stored at -80°C until further processing. Total RNA was isolated from liver tissue (n=8 mice per genotype) and subjected to microarray gene expression analysis.人类TRIB1（Tribbles同源蛋白1，Tribbles-homolog 1）基因位点的遗传变异与血浆脂质性状及冠状动脉疾病患病风险显著相关。本研究以动脉粥样硬化易感型低密度脂蛋白受体敲除（Ldlr-/-）背景小鼠为模型，分析Trib1基因敲除（Trib1-/-）对肝脏基因表达的影响。为此，我们将Trib1-/-小鼠与Ldlr-/-小鼠杂交，构建双基因敲除小鼠（Trib1-/-Ldlr-/-），并喂食含0.02%胆固醇、4.3%脂肪的半合成改良AIN76饲料。小鼠饲养至20周龄时，处死Trib1-/-Ldlr-/-小鼠及Trib1+/+Ldlr-/-对照小鼠，采集肝脏组织并经液氮快速冷冻后，于-80℃低温保存以待后续处理。从肝脏组织中提取总RNA（每种基因型设置8只小鼠作为生物学重复），随后开展基因芯片表达谱分析。