Gene expression in livers of Lep ob/ob (ob/ob) mice v.s. wild type C57BL/6J mice

To identify novel transcriptional factors involved in dysfunctional hepatic lipids homeostasis in obesity, mRNA microarray analysis were performed to of livers of ob/ob mice, a widely used obese model, and C57BL/6J control mice. Chow-fed ob/ob and C57BL/6J mice were housed in a 12 h of light and 12 h of dark cycle and fed ad libitum a regular chow diet. Mice were sacrificed at 16:00 (Zeitgeber Time 8 during light phase). Total RNA was prepared from each liver using TRIzol (Invitrogen). Equal aliquots of total RNA from each of four mouse livers in each group were pooled and used for biotin labeling as described in the Affymetrix technical bulletin. Then the transcriptional profiles of samples were probed using the Gene-Chip Mouse Genome 430 2.0 arrays.为鉴定参与肥胖相关肝脏脂质稳态失调的新型转录因子，本研究以广泛应用的肥胖模型ob/ob小鼠及C57BL/6J野生型对照小鼠为对象，对其肝脏组织开展mRNA微阵列分析。正常饲料喂养的ob/ob小鼠与C57BL/6J小鼠均饲养于12 h光照/12 h黑暗的光周期环境中，并自由采食常规维持饲料。于16:00（光照期授时时间8，Zeitgeber Time 8）处死实验小鼠。采用TRIzol试剂（Invitrogen公司）从每份肝脏样本中提取总RNA。将每组4只小鼠肝脏的总RNA等体积混合后，按照Affymetrix官方技术手册中的方法进行生物素标记。随后使用Gene-Chip小鼠基因组430 2.0芯片对样本的转录组表达谱进行杂交检测。