To explore the predictive utility of serum myoglobin (Mb) and lactate dehydrogenase (LDH) for acute kidney injury (AKI) secondary to rhabdomyolysis (RM) in severe heatstroke patients. A retrospective analysis of 58 RM patients with severe heatstroke at Shanghai Ninth People's Hospital from June 2019 to May 2022 was conducted. Patients were categorized into AKI and non-AKI groups. Laboratory indices were compared, and receiver operating characteristic (ROC) curves were used to assess the predictive value of serum biomarkers for AKI. Creatine kinase, Mb, LDH, creatinine, and blood urea nitrogen levels were significantly higher in the AKI group (P<0.05). Serum Mb and LDH were positively correlated with serum creatinine (r=0.6772 and r=0.6816, respectively; P<0.05). The area under the ROC curve (AUC) for serum Mb was 0.6692 (95% CI: 0.5253-0.8131) with a cut-off of 1024 ng/ml, while for LDH it was 0.8277 (95% CI: 0.7182-0.9371) with a cut-off of 1342 U/L. Combining serum Mb and LDH improved the AUC to 0.9116 (95% CI: 0.8219-1.001). Serum Mb and LDH levels are elevated in RM-induced AKI following severe heatstroke, and their combination offers substantial predictive value for AKI in these patients.