In this study, we found that spermine (Spm) and spermidine (Spd) injection for 6 weeks could prevent 24-month old rats heart dysfunction，improve mitochondrial function and down-regulate apoptosis. Using iTRAQ tools, we identify 75 mitochondrial proteins of statistically significant alteration in aging hearts, which mainly participate in important mitochondrial physiological activity, such as metabolism, translation, transport, apoptosis and oxidative phosphorylation.