Abstract Background End‐stage renal disease (ESRD) is the final stage of chronic kidney disease (CKD). Aims We aimed to analyze the expression differences of serum thrombomodulin (TM), platelet‐activating factor (PAF), and P‐selectin (CD62P) in patients with autologous arteriovenous fistula (AVF) and the correlation with vascular access function. Methods The case data were retrospectively analyzed. Moreover, 160 patients with AVF maintenance hemodialysis were selected as the AVF group, and 150 healthy participants were selected as the healthy control group. According to the function of vascular access, patients in the AVF group were divided into Group A (n = 50, after the first establishment of AVF), Group B (n = 64, normal vascular access function after hemodialysis treatment), and Group C (n = 46, vascular access failure). Pearson analysis was conducted to explore the correlation between serum TM, PAF, CD62P content, and vascular pathological examination indicators, to evaluate the value of TM, PAF, and CD62P levels in predicting vascular access failure in patients with AVF. Results and Discussion The serum levels of TM, PAF, and CD62P were positively correlated with the expressions of CD68 and MCP‐1, respectively (p