We reported that application of ethanol with lipopolysaccharide (LPS) and carbon tetrachloride (CCl4) enhanced alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level. In the current experi-ment, the protective effect of treadmill running on liver injury caused by ethanol with LPS and CCl4 in mice was studied. Liver injury severity was determined by measuring ALT and AST level in the blood. Terminal de-oxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining, immunohistochemistry for caspase-3, and Western blotting for Bcl-2-associated X protein (Bax) and B-cell lymphoma 2 (Bcl-2) were performed to indicate hepatocyte apoptosis. In addition, to understand the mechanism, 5’-adenosine monophosphate-activated protein kinase (AMPK) phosphorylation was studied by Western blotting. Treadmill ex-ercise ameliorated ethanol with LPS and CCl4-mediated elevation of ALT and AST level. Treadmill exercise suppressed ethanol with LPS and CCl4-mediated elevation of the TUNEL-positive cell number and cleaved caspase-3 expression. Treadmill exercise suppressed ethanol with LPS and CCl4-mediated elevation of Bax expression and increased Bcl-2 ex-pression suppressed by application of ethanol with LPS and CCl4. Tread-mill exercise enhanced AMPK phosphorylation which was suppressed by application of ethanol with LPS and CCl4. Treadmill exercise has the effect of reducing liver damage caused by alcohol and or drug addiction.