Aim To observe the anti-senescence effects of aspirin on activity of dimethylarginine dimethylaminohydrolase (DDAH)-asymmetric dimethylarginine (ADMA) system and caveolin-1 protein expression in human umbilical vein endothelial cells (HUVEC) exposed to high glucose condition and explore the mechanism of aspirin of anti-senescence. Methods The human umbilical vein endothelial cells (HUVEC) were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5.5 mmol/L glucose as normal level, 33 mmol/L glucose as high glucose, aspirin (0.01～1 mmol/L) with high glucose and 300 μmol/L L-NAME was added to the culture medium when needed for 48 hours. The activity of DDAH were reflected by ADMA concentration determined by high-performance liquid chromatography. The level of intracellular reactive oxygen species (ROS) was monitored by flow cytometry. Caveolin-1 protein expressions were analyzed by Western blot. Results After the endothelial cells were treated with high glucose concentration for 48 hours, the number of SA-β-gal positive cells, the level of ROS, ADMA and caveolin-1 protein were increased significantly. While, the activity of DDAH was decreased dramatically(P<0.05). All these changes were reversed by aspirin (0.01～1 mmol/L) remarkably in a dose-dependent manner (P<0.05). However, all the effects of aspirin on senescence were completely inhibited by L-NAME, the NOS inhibitor(P<0.05). Conclusion The anti-senescent effects of aspirin were fulfilled by inhibiting caveolin-1 protein expression and regulating the activity of DDAH-ADMA system.