A clinically-relevant mouse model that displays hemorrhage exacerbates tourniquet-induced acute kidney injury

Balamurugan Packialakshmi; David M Burmeister; Joseph A Anderson; Judah Morgan; Georgetta Cannon; Juliann G Kiang; Yuanyi Feng; Sang Lee; Ian J Stewart; Xiaoming Zhou

doi:10.3389/fphys.2023.1240352

A clinically-relevant mouse model that displays hemorrhage exacerbates tourniquet-induced acute kidney injury

Front Physiol. 2023 Nov 8:14:1240352. doi: 10.3389/fphys.2023.1240352. eCollection 2023.

Authors

Affiliations

¹ Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, United States.
² Department of Laboratory Animal Resources, Uniformed Services University of the Health Sciences, Bethesda, MD, United States.
³ Internal Medicine Residency Program at Madigan Army Medical Center, Joint Base Lewis-McChord, Tacoma, WA, United States.
⁴ Armed Forces Radiobiology Research Institute, Uniformed Services University of the Health Sciences, Bethesda, MD, United States.
⁵ Department of Pharmacology and Molecular Therapeutics, Uniformed Services University of the Health Sciences, Bethesda, MD, United States.
⁶ Department of Biochemistry and Molecular Biology, Uniformed Services University of the Health Sciences, Bethesda, MD, United States.

Abstract

Hemorrhage is a leading cause of death in trauma. Tourniquets are effective at controlling extremity hemorrhage and have saved lives. However, tourniquets can cause ischemia reperfusion injury of limbs, leading to systemic inflammation and other adverse effects, which results in secondary damage to the kidney, lung, and liver. A clinically relevant animal model is critical to understanding the pathophysiology of this process and developing therapeutic interventions. Despite the importance of animal models, tourniquet-induced lower limb ischemia/reperfusion (TILLIR) models to date lack a hemorrhage component. We sought to develop a new TILLIR model that included hemorrhage and analyze the subsequent impact on kidney, lung and liver injuries. Four groups of mice were examined: group 1) control, group 2) hemorrhage, group 3) tourniquet application, and group 4) hemorrhage and tourniquet application. The hemorrhagic injury consisted of the removal of 15% of blood volume through the submandibular vein. The tourniquet injury consisted of orthodontic rubber bands applied to the inguinal area bilaterally for 80 min. Mice were then placed in metabolic cages individually for 22 h to collect urine. Hemorrhage alone did not significantly affect transcutaneous glomerular filtration rate (tGFR), blood urea nitrogen (BUN) or urinary kidney injury molecule-1 (KIM-1) levels. Without hemorrhage, TILLIR decreased tGFR by 46%, increased BUN by 162%, and increased KIM-1 by 27% (p < 0.05 for all). With hemorrhage, TILLIR decreased the tGFR by 72%, increased BUN by 395%, and increased urinary KIM-1 by 37% (p < 0.05 for all). These differences were statistically significant (p < 0.05). While hemorrhage had no significant effect on TILLIR-induced renal tubular degeneration and necrosis, it significantly increased TILLIR-induced lung total injury scores and congestion, and fatty liver. In conclusion, hemorrhage exacerbates TILLIR-induced acute kidney injury and structural damage in the lung and liver.

Keywords: ischemia/reperfusion; liver injury; lower limb; lung injury; rhabdomyolysis; systemic inflammation.

Grants and funding

This work was supported in part by the Departments of Medicine and Surgery at the Uniformed Services University, the Uniformed Services University Sanford Endowment Fund, the United States of America Department of Defense grants W81XWH-22-2-0068 and MED-83-12715.