Establishment and Evaluation of a Diabetic Rat Model with Lower Limb Arteriosclerosis Obliterans

Summary

Lower extremity arteriosclerosis obliterans is a common complication of diabetes mellitus. Here, we present a novel animal modeling method that simulates the progression of atherosclerotic occlusive disease by injecting ethanol into blood vessels to disrupt vascular structure, thereby providing a new platform for the treatment of vascular diseases.

Abstract

This study aimed to establish and validate a novel rat model of lower extremity arteriosclerosis obliterans (ASO) in diabetic rats. Unlike traditional femoral artery ligation, we injected 95% ethanol (0.1 mL) into the external iliac artery. Postoperatively, arterial occlusion and hindlimb ischemia were assessed on days 1, 3, 7, and 14 using Doppler ultrasound, thermal imaging, and wet muscle weight measurement. Histological analysis of blood vessels and muscle tissues was performed to evaluate the model's success rate and stability. High-concentration ethanol (95%) induces vascular endothelial cell damage, leading to femoral artery occlusion. Compared to the control group, Doppler ultrasound revealed a progressive reduction in arterial diameter and increased flow velocity in the affected limb. Thermal imaging demonstrated decreased limb temperature, while muscle weight measurement showed a significant reduction in the ipsilateral muscle mass. Histopathological analysis confirmed arterial lumen occlusion, decreased microvascular density, and progressive lesion exacerbation over time. In conclusion, iliac artery injection of 95% ethanol effectively replicates the pathophysiological characteristics of lower limb arterial occlusion, providing a novel model for studying arteriosclerosis obliterans in diabetic rats.

Introduction

Diabetic lower extremity arterial occlusion (DLEAO) is a leading cause of disability and mortality in diabetic patients, severely impairing quality of life and imposing a substantial socioeconomic burden¹. The pathogenesis of diabetic arterial disease involves complex interactions among hyperglycemia, dyslipidemia, chronic inflammation, and vascular dysfunction². Animal models have been instrumental in elucidating these mechanisms, providing critical insights for therapeutic development. Animal models serve as valuable tools for simulating the pathophysiology of peripheral arterial disease (PAD) in human diabetic patient....

Protocol

This study was conducted under a protocol approved by the Animal Ethics Committee of Yantai Yuhuangding Hospital. This study utilized 40 male Sprague-Dawley rats (weight: 250-300 g, age: 6 weeks) housed under a 12 h light/dark cycle. Starting at 7 weeks of age, they were fed a high-fat, high-sucrose diet¹⁴^,¹⁵ ad libitum, with free access to water for 8 weeks. At 4 weeks into the diet, diabetes was induced via streptozotocin (STZ) injection¹⁶. After 8 weeks, successful establishment of the diabetic model was confirmed prior to subsequent experiments. Hindlimb arterial occlusion surgery was performed on 14-week-o....

Results

This study utilized intra-arterial ethanol injection to establish a model in diabetic SD rats (n = 40). The model was characterized by an 85% success rate (33/40), based on ultrasonic confirmation of reduced contralateral arterial diameter and blood flow velocity within 14 days post-injection. A model-related mortality rate of 8% (5/40) was recorded, predominantly during the initial period. While ethanol's cytotoxicity poses a risk of non-specific tissue damage such as gangrene, careful monitoring confirmed that the .......

Discussion

In summary, we have successfully established a rat model of progressive arterial occlusion that closely mimics the clinical manifestations observed in diabetic patients. Consistent evaluations of hindlimb hypoperfusion were achieved through ultrasonographic measurements and muscle wet weight assessments for periods exceeding two weeks. Ligation of the femoral artery induced superficial vascular occlusion, while allowing blood recirculation through the intact deep venous system¹⁷. When ligation is .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
GraphPad version 8.0.1			RRID:SCR_002798
Phone-based Infrared Thermometer	HIKMICRO		HM-TJP10B-3AMF
Sprague-Dawley rat	Jinan Pengyue Experimental Animal Breeding Co., Ltd.		250 g
Ultrasound machine	Vevo 2100