Background: This study aimed to evaluate the relationship between serum irisin and renal tubular biomarkers in patients with chronic kidney disease (CKD) undergoing maintenance hemodialysis. Methods: A prospective and observational study was conducted involving 50 patients with end-stage renal disease on regular hemodialysis. Blood samples were collected before and after a single dialysis session to assess the serum levels of irisin, urea, creatinine, neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), and liver-type fatty acid-binding protein (L-FABP) using the enzyme-linked immunosorbent assay and standard biochemical techniques. Results: Urea and creatinine levels significantly decreased post-dialysis, confirming effective clearance. Tubular injury markers (NGAL, KIM-1, and L-FABP) also showed notable reductions. Interestingly, serum irisin demonstrated a mild increase after dialysis, deviating from the trend seen with other biomarkers. A moderate inverse correlation was observed between irisin and tubular biomarkers, suggesting a potential protective or regulatory role. Conclusion: Serum irisin may provide unique insights into renal tubular health, distinct from traditional clearance markers. Its relative stability and inverse association with markers of injury position it as a candidate biomarker for assessing renal stress and recovery dynamics during hemodialysis. © 2025 Elsevier B.V., All rights reserved.