Cerebral Ischemic Coma Model Induced by Modified Four-Vessel Occlusion

Summary

This protocol describes the process for inducing a cerebral ischemic coma model using a modified four-vessel occlusion method.

Abstract

Coma caused by cerebral ischemia is the most serious complication of cerebral ischemia. Four-vessel occlusion can establish a cerebral ischemic coma model for disease research and drug development. However, the commonly used four-vessel occlusion method mainly involves inserting an electrocoagulation pen into the bilateral pterygoid foramen of the first cervical vertebra behind the neck to electrocoagulate the vertebral arteries. This process carries the risk of incomplete electrocoagulation, bleeding, and damage to the brainstem and spinal cord. Twenty-four hours after surgery, re-anesthetized rats undergo carotid artery ligation in front of the neck. Two surgeries expose the rats to a higher risk of infection and increase the experimental period. In this study, during a single surgical procedure, an anterior cervical incision was used to locate the key site where the vertebral artery penetrates the first cervical vertebra. The bilateral vertebral arteries were electrocauterized under visual conditions, while the bilateral common carotid arteries were separated to place loose knots. When the limbs of the rats began twitching, the bilateral common carotid arteries were quickly ligated to induce ischemic coma. This method can avoid the risk of infection caused by two surgical operations and is easy to perform with a high success rate, providing a useful reference for relevant practitioners.

Introduction

Ischemic brain injury is the most common brain injury in clinical practice, accounting for approximately 75% of cerebrovascular disease cases. Ischemia can lead to severe secondary brain injuries and diseases^1,2, and coma is the most severe symptom caused by ischemic hypoxic brain injury. It is also the final pathway for many critical conditions³. Coma is a critical and severe illness in clinical practice that is difficult to manage⁴. The longer the coma lasts, the greater the potential danger. Prompt awakening is the primary goal in preventing the deterioration ....

Protocol

The experimental protocol was conducted in accordance with the requirements of the Use of Laboratory Animals and Institutional Animal Care and Use Committee at Foshan University (Record number: 2023-643656). Male Sprague Dawley (SD) rats (200 g ± 20 g, 6-8 weeks old) were used for this study. All animal research data have been written up in accordance with the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines. The details of the reagents and equipment used in the study are listed in the Tabl.......

Discussion

Four-vessel occlusion induces global ischemic and hypoxic brain injury, which can simulate acute coma, cardiac arrest, asphyxia, shock, severe arrhythmia, and other critical clinical conditions caused by cerebral ischemia in clinical practice. Meanwhile, four-vessel occlusion can lead to damage mainly in the hippocampus^17,18, which is the primary functional brain area responsible for cognitive memory^19,20.......

Materials

Name	Company	Catalog Number	Comments
16 channel microfiber photoelectrode array	Jiangsu Yige Biotechnology Co., Ltd	2605
4-0 Surgical suture	Nantong Holycon Medical Devices Co.,Ltd.	B-104
6-0 Surgical suture	Ningbo MEDICAL Needle Co., Ltd.	JM1216-742417
EEG electrode	Kedou Brain machine Technology Co., LTD	KD-EEGEMG
Electrocoagulation pen	CONPUVON Company	465
Lunion Stage Automatic Sleep Staging System	Shanghai Lulian Intelligent Technology Co., Ltd.	1336
Miniature hand-held skull drill	Rayward Life Technology Co., Ltd	87001
Penicillin sodium	Chengdu Kelong Chemical Co., Ltd.	17121709-2
SD rats	SPF ( Beijing ) Biotechnology Co.,Ltd.	180-220g
Skull nail	GLOBALEBIO,LTD	/
Stereotaxic instrument	Rayward Life Technology Co., Ltd	68801
Zoletil 50	Vic Trading (Shanghai) Co., LTD	BN 88SHA