Effect of Mesenchymal Stem Cell-Derived Extracellular Vesicles Induced by Advanced Glycation End Products on Energy Metabolism in Vascular Endothelial Cells

Introduction: Advanced glycation end products (AGEs) play a critical role in the development of vascular diseases in diabetes. Although stem cell therapies often involve exposure to AGEs, the impact of this environment on extracellular vesicles (EVs) and endothelial cell metabolism remains unclear.

Methods: Human umbilical cord mesenchymal stem cells (MSCs) were treated with either 0 ng/ml or 100 ng/ml AGEs in a serum-free medium for 48 hours, after which MSC-EVs were isolated. The EVs were characterized by morphology, particle size, and protein markers of MSC-EVs, and microRNA (miRNA) sequencing was performed to identify differentially expressed miRNAs. MSC-EVs were cocultured with human umbilical vein endothelial cells (HUVECs) to assess effects on cell viability, metabolic activity, oxidative stress, and antioxidant capacity. Tube formation and glucose transporter protein analyses were conducted to evaluate the angiogenic ability and glucose metabolism capacity.

Results: MSC-EVs ranged from 30 to 150 nm, which is consistent with exosomal properties. AGEs treatment reduced MSC viability but had minimal effect on EV morphology and protein markers. miRNA sequencing showed downregulation of hsa-miR-223-3p and hsa-miR-126-3p_R-1, with upregulation of hsa-miR-574-5p, implicating changes in glycolytic and oxidative phosphorylation pathways. MSC-EVs treated with AGEs decreased HUVEC viability (P < 0.05), pH (P < 0.05), adenosine triphosphate (ATP) metabolism (P < 0.05), glucose metabolism (P < 0.05), while enhancing glycolysis processes, including glycolytic activity, capacity, and reserve (P < 0.05). This likely resulted from impaired mitochondrial function, including reduced ATP production, maximal respiration, basal respiration, and spare respiratory capacity (P < 0.05), or increased reactive oxygen species (ROS) (P < 0.05) and glucose-6-phosphate dehydrogenase (G6PD) activity (P < 0.05). In addition, AGEs reduced glucose transporter types 1, 3, and 4 (GLUT1, GLUT3, GLUT4), and synthesis of cytochrome c oxidase 2 expression (P < 0.05), along with angiogenic capacity (P < 0.05) in HUVECs.

Conclusion: Exposure to AGEs diminishes the therapeutic potential of MSC-derived EVs by disrupting energy metabolism and promoting metabolic reprogramming in endothelial cells. These findings suggest that adjusting the dosage or frequency of MSC-EVs may enhance their efficacy for treating diabetes-related vascular conditions. Further research is warranted to evaluate AGEs' broader impact on various cell types and metabolic pathways for improved exosome-based therapies.