Hepatic gene expression profiles during fed–fasted–refed state in mice [mRNA-seq]

Regulation of nutrient status during fasting and refeeding plays an important role in maintaining metabolic homeostasis in the liver. Thus, we investigated the impact of the physiological fed–fasted–refed cycle on hepatic gene expression in nutrient-sensitive mice. We performed transcriptomic analysis of liver samples in fed, fasted and refed groups of mice. Through mRNA-sequencing (RNA-Seq) and miRNA-Seq, we compared fasted and fed states (fasted versus fed cohort) as well as refed and fasted states (refed versus fasted cohort) to detect dynamic alterations of hepatic mRNA– miRNA expression during the fed–fasted–refed cycle The animal procedures were approved by the Animal Experiment Ethics Committees of Shanghai Jiao Tong University School of Medicine. Wild-type male C57BL/6J mice aged 8 weeks were purchased from Shanghai Laboratory Animal Company (SLAC, Shanghai, China). All mice were housed at 21±1°C with a humidity of 55±10% and 12- h light/12-h dark cycle in a specific-pathogen-free facility. After 2 weeks of acclimatization, mice were divided into three groups. The fasted group was fed a regular diet with subsequent fasting for 24 h. The refed group underwent 24 h fasting and was refed a fixed-calorie meal for 2 h. Mice were anesthetized with sodium pentobarbital (Nembutal, 80 mg/kg, i.p.) and killed during the fasted and refed states. Liver tissues were harvested and snap-frozen in liquid nitrogen for further analysis. RNA-Seq