Bilateral and Unilateral Renal Ischemia‐Reperfusion Injury Model in Rats

Renal ischemia-reperfusion injury (IRI), one of the most important causes of acute kidney injury, is a significant health problem that can result in acute kidney failure after surgical procedures in which blood flow to the kidney is temporarily interrupted. Although numerous studies have been conducted to prevent renal IRI damage, this problem remains unresolved due to its complex nature. During IRI, both ischemia and reperfusion contribute to damage, with oxidative stress, inflammation, and the activation of apoptotic mechanisms playing significant roles in exacerbating the damage. This article provides comprehensive protocols for inducing renal IRI in rats, including both bilateral and unilateral models. These protocols can be used in studies designed to understand the cellular and molecular mechanisms of renal IRI and exploring potential therapeutic strategies. When the described procedure is carefully applied in experimental IRI models, the mortality rate in animals will be lower. Furthermore, we outline opportunities for investigating oxidative stress, inflammatory and apoptotic pathways, and highlight how these analyses may contribute to preclinical therapeutic studies. We also provide recommendations on which analyses should be performed in such studies. Specifically, information is provided on which kidney damage biomarkers should be examined in samples obtained from animals. Additionally, we explain which investigations can be performed on apoptotic and inflammatory pathways involved in kidney damage. In summary, this study integrates bilateral and unilateral approaches and provides a detailed methodological framework for mechanistic and therapeutic research on renal ischemia-reperfusion injury. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Bilateral kidney ischemia-reperfusion in rats Alternate Protocol: Unilateral kidney ischemia-reperfusion in rats Basic Protocol 2: Analyses performed on kidney tissues and serum of rats after ischemia-reperfusion.