Aspirin Eugenol Ester Reduces H2O2-Induced Oxidative Stress of HUVECs via Mitochondria-Lysosome Axis

The oxidative stress of vessel endothelium is a major risk factor of cardiovascular disorders. Antioxidative stress drugs are widely used in cardiovascular therapy. Aspirin eugenol ester (AEE) is a new pharmaceutical compound synthesized by esterification reaction of aspirin with eugenols and possesses antioxidative activity. The present study was designed to investigate the mechanism how AEE protects human umbilical vein endothelial cells (HUVECs) from H₂O₂-induced oxidative stress. H₂O₂ was given to the HUVECs with or without AEE pretreatment. Changes in the oxidative stress-related factors, including those related to the mitochondria-lysosome axis, were determined with Western blotting, cellular immunofluorescence, and Enzyme activity test. The results showed that, in the HUVECs, 300 μM H₂O₂ treatment significantly increased the Apoptosis rate, MDA concentration, Reactive Oxygen Species (ROS) production, mitochondrial membrane potential, expression of Bax and mature Cathepsin D (CTSD), and activity of CTSD and Caspase3 (Cas3) but decreased the expression of Bcl2 and lysosomal membrane stability, while in the HUVECs pretreated with AEE, the above changes caused by either the stimulatory or the inhibitory effect of H₂O₂ on the relevant factors were significantly reduced. AEE pretreatment significantly enhanced the activity of cellular superoxide dismutase and Glutathione Peroxidase in the HUVECs. Our findings suggest that AEE effectively reduced H₂O₂-induced oxidative stress in the HUVECs via mitochondria-lysosome axis.