Purpose: This study aimed to investigate changes in muscle damage during the course of a 217-km mountain ultramarathon (MUM). In an integrative perspective, inflammatory response and renal function were also studied. Methods: Six male ultra-runners were tested four times: pre-race, at 84 km, at 177 km, and immediately after the race. Blood samples were analyzed for serum muscle enzymes, acute-phase protein, cortisol, and renal function biomarkers. Results: Serum creatine kinase (CK), lactate dehydrogenase (LDH), and aspartate aminotransferase (AST) increased significantly throughout the race (P < 0.001, P < 0.001; P = 0.002, respectively), and effect size (ES) denoted a large magnitude of muscle damage. These enzymes increased from pre-race (132 ± 18, 371 ± 66, and 28 ± 3 U/L, respectively) to 84 km (30, 1.8, and 3.9-fold, respectively); further increased from 84 to 177 km (4.6, 2.9, and 6.1-fold, respectively), followed by a stable phase until the finish line. Regarding the inflammatory response, significant differences were found for C-reactive protein (CRP) (P < 0.001) and cortisol (P < 0.001). CRP increased from pre-race (0.9 ± 0.3 mg/L) to 177 km (243-fold), cortisol increased from pre-race (257 ± 30 mmol/L) to the 84 km (2.9-fold), and both remained augmented until the finish line. Significant changes were observed for creatinine (P = 0.03), urea (P = 0.001), and glomerular filtration rate (GFR) (P < 0.001), and ES confirmed a moderate magnitude of changes in renal function biomarkers. Creatinine and urea increased, and GFR decreased from pre-race (1.00 ± 0.03 mg/dL, 33 ± 6 mg/dL, and 89 ± 5 ml/min/1.73 m2, respectively) to 84 km (1.3, 3.5, and 0.7-fold, respectively), followed by a plateau phase until the finish line. Conclusion: This study shows evidence that muscle damage biomarkers presented early peak levels and they were followed by a plateau phase during the last segment of a 217-km MUM. The acute-phase response had a similar change of muscle damage. In addition, our data showed that our volunteers meet the risk criteria for acute kidney injury from 84 km until they finished the race, without demonstrating any clinical symptomatology.