Standardization of the Simple Methodology for Experimentally Induced Ischemic Stroke in Rat Models

Abstract

Stroke is a globally significant and devastating disease that requires prompt treatment. Animal models are commonly used to investigate stroke therapy, often through experimentally induced ischemic stroke (EIIS). However, challenges arise in implementing EIIS in animal models. The current study aimed to present a simple EIIS methodology for animal models. A total of 60 male Sprague-Dawley rats were randomly divided into five groups, namely Group 1 (sham-operated), Groups 2 to 5 (EIIS groups) with different duration of common carotid artery (CCA) ligation, including 1, 2, 4, and 8 hours, respectively. The ligation was performed on the CCA and its branches. Before the experiment, the rats were anesthetized, and the incision area was shaved and disinfected. The sagittal ventral midline was incised, with neck muscles retracted to expose the right CCA. The occlusion was performed on three sides of a carotid artery (common, external, and internal) using a simple interrupted suture. The occlusion of blood flow using ligation was performed at different times depending on the groups. After that, the CCA ligations were re-perfused by cutting the suture knot. The brain and blood were collected on days 1 and 7 after reperfusion. The results indicated that 4 and 8 hours of CCA ligation significantly impacted the general condition and neuro-deficit score. Moreover, 4 and 8 hours of CCA ligation could induce ischemic stroke by its capacity to cause infarction within the brain parenchyma and increase the platelet-to-white blood cell ratio, C-reactive protein, and De Ritis ratio. In contrast, 1 and 2 hours of CCA ligation did not significantly affect the observed parameters. It can be concluded that the EIIS using 4 and 8 hours of CCA ligation can be applied to induce ischemic stroke in rat models with consistent impacts on general conditions, neuro-deficit, hematology, and serology.