Tramadol and alcohol are among commonly abused drugs. Although there are po-tential dangers reported upon their mixing, there are no previous reports describingthis mixture's effects on the cardiovascular system (CVS). The aim was to study theeffects of mixed alcohol and tramadol on the CVS of adult male rats. Fifty rats weredivided into four groups: control, tramadol‐treated group, alcohol‐treated, andcoadministration groups. Tramadol caused a significant increases in creatine kinase‐MB, troponin I, malondialdehyde, protein carbonyl, 8‐hydroxy‐2′‐deoxyguanosine,and a significant decrease in total antioxidant capacity with histological alterationsin sections of the heart and aorta and a significant increase in the area% of collagenfibers while there was a nonsignificant difference in body weight, heart weight,heart weight/body weight ratio, lipid profile, tissue tumor necrosis factor‐αandinterferon‐γ, intermediate microfilament proteins (IFPs) {desmin, vimentin, con-nexin43} gene expression, mean area% of elastic fibers in aortic tissue and osteo-pontin expression in cardiac and aortic tissue. Alcohol treatment caused a significantchange in all the measured parameters and more damage in histological sections.The changes were highest in the coadministration group. There was a strong posi-tive correlation between the area% of collagen fibers and vimentin gene expression,and the area% of osteopontin expression was positively correlated to connexin43 incardiac and vascular tissue. Tramadol causes CVS injury mainly through oxidativestresses, while the alcohol effect is multifactorial; mixing both aggravates CVSinjury. The study also highlights the role of IFPs and osteopontin‐expression ininducing injury