Background Cholinesterase inhibitors are used for symptomatic treatment of mild to moderate Alzheimer's disease. These drugs have vagomimetic and anti-inflammatory properties that could be of interest also with respect to cardiovascular diseases. Aim The aim of the present work is to assess effects of pretreatment with donepezil hydrochloride, one of cholinesterase inhibitors, on acute myocardial infarction induced by coronary artery ligation in rats. Methods Rats were randomly divided into control, sham-operated, untreated-myocardial infarction, and donepezil hydrochloride-pretreated myocardial infarction groups. Donepezil hydrochloride was administered orally, 1mg/kg/day, for six days then acute myocardial infarction was induced in 6^th day by ligation of the left anterior descending coronary artery. ECG and blood pressure were recorded. Triphenyltetrazolium chloride (TTC) stained ventricular slices were examined. The plasma levels of tumor necrosis factor-α (TNF-α) and cardiac troponin I (cTnI) were assessed. Catalase activity and glutathione levels in the myocardium were determined. Results Coronary artery ligation increased T-wave voltage with appearance of pathological Q-wave, tachycardia, decreased mean arterial pressure and pale areas (infarcts) in TTC-stained ventricular slices. The plasma levels of TNF-α and cTnI were increased while the myocardial catalase activity and glutathione levels were decreased. In donepezil hydrochloride-pretreated group T-wave voltage and heart rate were decreased and pathological Q-wave disappeared with decrements in infarct size and plasma levels of TNF-α as well as cTnI, while catalase activity and glutathione levels were increased in relation to untreated myocardial infarction group. Conclusion Pretreatment with donepezil hydrochloride decreased myocardial infarct size. This effect could be related to its negative chronotropic, anti-inflammatory, and antioxidant actions.