Abstract: Objective  To establish a mouse  model of internal carotid arteriovenous fistula (AVF) and screen for differentially expressed proteins during AVF intimal hyperplasia and stenosis, and analyze the aberrant signaling pathways associated with this process.  Methods  A common carotid artery-external jugular vein AVF model was established in C57BL/6 mice using microsurgical techniques. The mice were divided into a sham operation group (n=10) and an AVF operation group (n=10). Vascular pathological changes in the AVF were evaluated. Tandem mass tag (TMT) quantitative proteomics analysis was used to identify differentially expressed genes and proteins between the two groups, and bioinformatics methods were applied to perform enrichment analysis of the differentially expressed proteins. For the cell experiment, human umbilical vein endothelial cells (HUVECs) were divided into a control group and a shear stress group, to detect reactive oxygen species (ROS) and mitochondrial morphology.  Results  In the animal experiment, compared with the sham operation group, the AVF operation group exhibited significant venous intimal hyperplasia, characterized by a mixture of collagen fiber proliferation and muscular layer components. Proteomic analysis identified a total of 595 differentially expressed proteins. Enrichment analysis revealed that these genes were primarily involved in biological processes such as mitochondrial homeostasis and energy metabolism, and were enriched in glycolipid metabolism pathways and the peroxisome proliferators-activated receptors (PPAR) signaling pathway.  In the cell experiment, ROS production in HUVECs in the shear stress group was higher than in the control group (t=8.840, P＜0.001). Mitochondrial area and mitochondrial length were lower in the shear stress group compared to the control group (t=5.926, 4.579, both P＜0.001), while mitochondrial superoxide levels were higher  (t=28.170, P＜0.001).  Conclusion  Mitochondrial oxidative stress injury plays a role in the process of intimal hyperplasia and stenosis in AVF.

Key words: Arteriovenous fistula, Intimal hyperplasia, Proteome sequencing, Oxidative stress