Introduction: Atherosclerosis is most common cause of acute myocardial infarction (MI) and can lead to increased oxidative stress and damage to macromolecules. The aim of this study was to determine the amount of 8-Hydroxydeoxyguanosine (8- OHdG) as a marker of damage to the DNA and the levels of heat shock protein HSP27 in acute myocardial infarction (MI) patients. Materials and Methods: In this case-control study, 30 patients with acute MI were enrolled. Serum levels of HSP27, 8-OHdG, cardiac troponin I (C-TnI), creatine kinase (CK-MB) and levels of total antioxidant and malondialdehyde were assessed. Data were analyzed by SPSS 16 and T-test analysis at the significant level of P≤0.05. Results: Data analysis showed that serum levels of HSP27 and 8-OHdG in acute MI patients peaked (8-OHdG=9.8±2.1, HSP27=81±3.1) after 48 hours of the attack showing a significant increase (HSP27=9.7±1.8, 8-OHdG=4.4±1.2) (P≤0.05) in comparison to healthy controls. Levels of the enzyme CK-MB 24 hours and CTnI and malondialdehyde after 48 hours after acute MI showed the highest values. Conclusion: Increase in markers of DNA damage and heat shock protein levels in MI patients, verify high levels of oxidative stress in MI, indicating that evaluation of changes in 8-OHdG and heat shock proteins is a valuable way to assess the level of damage to macromolecules during acute MI.