Mice fed 4% alcohol and 0.5% cholesterol LD causes greater hepatic lipid accumulation and increases liver tissue histological inflammation for feeding 5 weeks. (a and c) Liver damage directly reflected by H&E (x 40 and x 400). (b) Oil Red O staining shows the excessive cytoplasmic lipid accumulation (x 200). (d) Immunohistochemistry reflected the expression of TLR4 (x 400). (e) The data of TLR4 expression was semiquantitatively analysed as integrated option density (IOD) in positive area of the microphotograph. (f) Western blot reflected the expression of NF-κB and p-NF-κB. (g) Hepatic lipid accumulation directly reflected by the level of TC in liver. (a–f) Alcohol induced more severe liver injury and steatosis with cholesterol ingestion. Values were expressed as the mean ± SD (n=3), #P < 0.05; ##P < 0.01 versus NLG. (g) Values were expressed as the mean ± SD (n=12), #P < 0.05; ##P < 0.01 versus NLG; ∮P < 0.05; ∮∮P < 0.01 versus CLG.