Alcohol and HFD induce hepatic steatosis. (a) HFD and Alc+HFD increased liver triglyceride. Analysis of liver homogenates showed that HFD- and Alc+HFD-treated mice had significantly higher triglyceride per gram of liver compared with livers from Chow- and Alcohol-treated mice (n=5). (b) Alcohol and HFD increased serum triglyceride, cholesterol and HDL. Serum triglyceride concentration was significantly higher in mice treated with Alcohol compared with control. It was significantly reduced with HFD and Alc+HFD when compared with Alcohol alone. Serum from mice treated with HFD and Alc+HFD showed significantly higher cholesterol and HDL than those on Chow diet (n=7–8). (c) Alcohol and HFD altered expressions of lipid processing mRNAs favouring lipid accumulation in the liver (Q-PCR). Alc+HFD induced significant expression of SREBP-1 mRNA compared with all other treatment groups. Alcohol alone and Alc+HFD induced significant expression of SCD-1 mRNA compared with Chow. SCD-1 expression was significantly reduced in HFD compared with Alcohol alone and Alc+HFD. mRNA expression of PPARα was significantly induced in HFD and Alc+HFD from both Chow and Alcohol treatment groups. HFD significantly reduced ACOX1 mRNA expression compared with Chow and Alcohol, while Alc+HFD significantly reduced ACOX1 expression compared with Alcohol. mRNA expression of target genes was normalised using housekeeper Hsp90ab1 (n=5–6). Data are mean ±s.d. *P<0.05 from Chow, **P<0.01 from Chow, ****P<0.0001 from Chow; #P<0.05 from Alcohol, ##P<0.01 from Alcohol, ###P<0.001 from Alcohol, ####P<0.0001 from Alcohol; δP<0.05 from HFD, δδδP<0.001 from HFD. HDL, high density lipoprotein; SREBP-1, sterol response element binding protein-1; SCD-1, stearoyl-CoA desaturase-1; Proliferation peroxisome activated receptor α, PPARα; ACOX1, acyl-CoA oxidase 1.