These changes were followed by death or distress, necessitating euthanasia within 48 hours. Fifty percent of KO mice died within the first 6 days of initiating the 5% ethanol diet, whereas none died in the WT/ethanol group (Fig. 1A). Food intake was similar in the two EtOH groups, except for just before death

in the KO group (Fig. 1B). To avoid confounding results from animals in http://www.selleckchem.com/products/lee011.html extremis, we sacrificed the remaining mice after day 6 on 5% ethanol, and the experiments described below were performed on these mice. PF KO and WT mice appeared healthy and gained weight (data not shown). EtOH KO mice were hypoglycemic with 2-fold lower blood-glucose levels than WT mice (Fig. 1C) and had 10% lower body weight (Fig. 1D). EtOH KO mice had cachexia and severely depleted intra-abdominal fat, compared with the WT/ethanol group, likely representing a baseline defect in energy homeostasis and ethanol-induced acute illness and decreased food intake click here in KO mice (Fig. 1E; Supporting Fig. 1 24). There was no difference

in body temperature between the groups. We conclude from these results that KO mice are highly susceptible to systemic toxicity and death after short exposure to ethanol ingestion. Both groups of KO mice had lower liver weight (Supporting Fig. 2). However, only PF KO mice had a lower liver:body weight ratio, compared with the corresponding WT group (Supporting Fig. 3). On microscopic examination of the liver, EtOH KO mice exhibited severe micro- and

macrovesicular steatosis in all three zones of the liver lobule. In contrast, WT mice developed only mild (predominantly zone 2) microvesicular steatosis (Fig. 2A, upper panel). Similarly, Oil red O staining for neutral lipids confirmed the presence of increased hepatic steatosis in the KO/ethanol group (Fig. 2A, bottom panel). KO mice had approximately 5-fold higher alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels than WT mice on the ethanol diet (Fig. 2B,C). Biochemical assays revealed higher liver triglyceride and cholesterol levels in the KO/ethanol group, compared with WT mice (Fig. 2D,E). Serum triglyceride and total cholesterol levels were similar in WT and KO mice (data not shown). Thus, these results show that KO mice develop severe selleck chemical liver steatosis and moderate transaminase elevation on ethanol ingestion in a time period that causes only mild lipid accumulation and no change in liver injury tests in WT mice. Increased hepatic oxidative stress is an important mechanism of ethanol-mediated liver injury, and lipid peroxidation (LPO) is used as an indicator of oxidative stress in tissues. Therefore, we performed an assay for malondialdehyde (MDA) levels as an indicator of LPO in the liver. KO mice had higher hepatic MDA levels than WT mice on the ethanol diet (Fig. 3A).