ACSL3/GABARAPL2 Ameliorates Vascular Endothelial Cell Aging and Injury Through Protective Autophagy to Alleviate Ferroptosis.

Vascular endothelial cell (VEC) dysfunction is a key contributor to vascular diseases. Oxidative stress-induced ferroptosis accelerates VEC aging and injury, while protective autophagy mitigates this damage by clearing damaged organelles and reducing oxidative stress. GABARAPL2, a critical autophagy-related protein, and ACSL3, a regulator of lipid metabolism and ferroptosis, are emerging as interconnected mediators of cellular protection. However, their precise roles in oxidative stress-induced VEC aging and injury remain unclear. This study aimed to investigate how GABARAPL2 and ACSL3 regulate autophagy and ferroptosis to protect VECs. Human umbilical vein endothelial cells (HUVECs) were treated with hydrogen peroxide to establish a cellular oxidative stress model. Gene expression was analyzed through Western blot and immunofluorescence, while cell viability, apoptosis, reactive oxygen species (ROS), Fe levels, and mitochondrial function were assessed. GABARAPL2 and ACSL3 functions were investigated through knockdown and overexpression experiments, and autophagy and ferroptosis markers were evaluated. Knockdown of GABARAPL2 and ACSL3 significantly increased aging markers (P21, P53, PAI-1) and reduced HUVEC viability. Overexpression of GABARAPL2 enhanced protective autophagy, reduced ROS and Fe levels, and alleviated ferroptosis under oxidative stress. GABARAPL2 upregulated ACSL3 expression, which further promoted autophagy and inhibited ferroptosis. Autophagy inhibition reversed these protective effects, indicating that GABARAPL2/ACSL3-mediated autophagy plays a key role in mitigating ferroptosis and oxidative stress. GABARAPL2/ACSL3 improves VEC aging and injury by promoting protective autophagy and reducing ferroptosis, thereby improving cellular viability under oxidative stress. These findings offer a potential therapeutic target for diseases associated with endothelial dysfunction.