Ferritinophagy Is Involved in the Advanced Glycation End Products-Induced Ferroptosis of Vascular Endothelial Cells in Diabetes Mellitus.

This study investigated the role of advanced glycation end products (AGEs) and ferritinophagy-dependent ferroptosis in the pathogenesis of diabetic atherosclerosis obliterans. We investigated the effects of AGEs on ferroptosis and ferritinophagy in HUVECs. The methods employed included the CCK-8 assay, reactive oxygen species (ROS) determination, Fe2+ measurement, Western blot, quantitative PCR (qPCR), transmission electron microscopy (TEM) and immunofluorescence. Additionally, we established type 2 diabetes rat models and atherosclerosis obliterans (ASO) models. Blood flow in the rats' lower limbs was measured using the RFLSI III Laser Speckle Imaging System. Furthermore, levels of glutathione (GSH) and malondialdehyde (MDA) were measured to evaluate oxidative stress and lipid peroxidation. Upon exposure of HUVECs to AGEs, a reduction in cell viability was observed, accompanied by a significant increase in iron ion and MDA levels, and a notable decrease in GSH levels, indicating the induction of ferroptosis. The addition of ferrostatin-1 to AGEs-treated HUVECs mitigated ferroptosis. TEM, immunofluorescence and Western blot analyses confirmed that AGEs can induce ferritinophagy in HUVECs. Furthermore, compared to the human serum albumin (HSA) group, the AGEs-treated group exhibited significantly elevated levels of reactive oxygen species (ROS) and MDA, along with a marked reduction in GSH levels. These alterations were reversed by the addition of MitoTEMPOL to the AGEs group. Animal experiments further confirmed that the clearance of intracellular ROS alleviated ferritinophagy-dependent ferroptosis. This study provides an insight into therapeutic strategies for alleviating ASO in DM by interfering with the ferritinophagy-dependent ferroptosis due to AGEs accumulation.