Laboratory mice are standardly housed around 23ºC, setting them under chronic cold stress. Metabolic changes of liver in mice housed under thermoneutral, standard and cold temperatures remain unknown. Here we isolated lipid droplets and mitochondria from their livers for comparative proteomic study to investigate the changes. According to proteomic analysis, mitochondrial TCA cycle and retinol metabolism were enhanced while oxidative phosphorylation was no obviously affected under cold condition, suggesting liver mitochondria may increase TCA cycle capacity in biosynthetic pathways as well as retinol metabolism to help liver to adapt. Based on proteomic and immunoblotting results, Perilipin 5 and major urinary proteins were increased significantly while mitochondrial pyruvate carrier was decreased dramatically under cold condition, indicating a role they may play.