Mouse abdominal aortic aneurysm model construction method

Abstract

The invention relates to the technical field of animal model construction, and discloses a mouse abdominal aortic aneurysm model construction method. The construction method comprises the step that in a 18-month-old mouse, a sterile cotton piece soaked with elastinase is used for wrapping the kidney subabdominal aorta to induce formation of the abdominal aortic aneurysm. The mouse model can better reflect the degenerative change of the aortic wall of an elderly patient, better fits the clinical manifestation in the development speed and severity of aortic aneurysm, can more stably simulate the development process of the abdominal aortic aneurysm of the elderly patient, can be used as an effective tool for studying senescence-related aneurysm diseases, and has a wide application prospect. The method is helpful for revealing acceleration and aggravation processes of the elderly individuals in pathological development, and provides a more targeted theoretical basis and experimental support for designing and evaluating a new treatment method.

Description

Technical Field

[0001] The present application relates to the technical field of animal model construction, and more specifically, to a method for constructing a mouse abdominal aortic aneurysm model. Background Art

[0002] Abdominal aortic aneurysm (AAA) is a common vascular disease that primarily occurs in the elderly. It is characterized by abnormal dilation of the aortic wall, which can lead to rupture and be life-threatening. Establishing a stable and reliable animal model is crucial for in-depth understanding of the pathogenesis of this disease and exploring potential treatments.

[0003] Currently, the commonly used method for establishing a mouse abdominal aortic aneurysm model is to induce aneurysm formation in young mice (e.g., 6-8 weeks old) using porcine pancreatic elastase (PPE). However, the existing PPE-induced model has the following drawbacks:

[0004] (1) Large pathological differences from elderly patients: The physiological state of young mice is significantly different from that of elderly patients. The structure and function of their aortic walls are relatively young, and they cannot accurately simulate the pathological characteristics of elderly abdominal aortic aneurysm patients;

[0005] (2) Poor model stability: The abdominal aortic aneurysm model in young mice after PPE induction is relatively unstable and prone to complications such as aneurysm rupture, which is inconsistent with the clinical characteristics of abdominal aortic aneurysms in elderly patients, which are slow expansion and gradual aggravation.

[0006] (3) Limited research value: The model constructed using young mice has limitations in studying elderly-related abdominal aortic aneurysm diseases and cannot deeply explore the role of aging-related factors in the occurrence and development of abdominal aortic aneurysms.

[0007] In summary, the existing young mouse PPE-induced model has obvious deficiencies in simulating the characteristics of elderly human AAA disease. There is an urgent need to develop a new animal model that is closer to clinical pathological characteristics to improve the reliability of research and its translational medical value. Summary of the Invention

[0008] The invention purpose of this application is to better simulate the pathogenesis of abdominal aortic aneurysm in elderly humans and provide a more accurate basis for clinical diagnosis and treatment.

[0009] To achieve the above-mentioned purpose of the invention, the present application provides a method for constructing a mouse abdominal aortic aneurysm model, which adopts the following technical solutions:

[0010] In a first aspect, the present application provides a method for constructing a mouse abdominal aortic aneurysm model, comprising the steps of inducing abdominal aortic aneurysm formation in 18-month-old mice by wrapping the infrarenal abdominal aorta with a sterile cotton sheet soaked in elastase.

[0011] Furthermore, the 18-month-old mice are 18-month-old male C57BL / 6J mice with an average body weight of 30 to 40 g.

[0012] Furthermore, the sterile cotton piece is soaked with 35 to 45 uL of 10 mg / mL elastase.

[0013] Furthermore, the infrarenal abdominal aorta is wrapped with a sterile cotton sheet soaked with elastase for 30 to 50 minutes.

[0014] Furthermore, the construction method comprises the following steps: using 18-month-old male C57BL / 6J mice with an average weight of 30 to 40 g, anesthetizing them with isoflurane, fixing the mice in a supine position, placing the nose and mouth of the mice in an anesthesia mask, and adjusting the amount of isoflurane to 1.5% to 2.0%; after anesthesia, removing the abdominal hair, disinfecting and laying a drape; cutting the abdominal skin and peritoneum, peeling off the intestine, exposing and separating the infrarenal abdominal aorta, using sterile gloves of appropriate size to separate the infrarenal abdominal aorta from the surrounding tissues, and then wrapping the infrarenal abdominal aorta with a sterile cotton pad soaked in 40 uL, 10 mg / mL elastase to temporarily close the abdominal cavity; removing the cotton pad and gloves after 40 minutes, gently wiping away the residual fluid in the abdominal cavity with a sterile cotton swab dipped in normal saline, suturing the peritoneum and skin separately with 5-0 surgical sutures, and transferring the mice to a cage after they wake up and continue feeding.

[0015] In a second aspect, the present application provides a mouse abdominal aortic aneurysm model obtained by the above-mentioned construction method.

[0016] In a third aspect, the present application provides the use of the above-mentioned mouse abdominal aortic aneurysm model in screening drugs for treating and / or preventing aging-related aneurysm diseases.

[0017] Furthermore, the aging-related aneurysm disease includes abdominal aortic aneurysm in elderly patients.

[0018] In summary, this application has the following beneficial effects:

[0019] This application uses aged mice, which can better reflect the degenerative changes in the aortic wall of elderly patients, such as the reduction of smooth muscle cells, rearrangement of collagen fibers, and destruction of elastic fibers.

[0020] The elderly mouse model of the present application is more consistent with clinical manifestations in terms of the development speed and severity of aortic aneurysms, and can more stably simulate the development process of elderly abdominal aortic aneurysms.

[0021] The PPE model of elderly mice in this application can be used as an effective tool to study aging-related aneurysmal diseases, helping to reveal the accelerated and aggravated process of pathological development in elderly individuals, and providing a more targeted theoretical basis and experimental support for the design and evaluation of new treatment methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 : Pathological differences between young mice (6-8 weeks) and old mice (18 months) in the PPE-induced abdominal aortic aneurysm model in mice. DETAILED DESCRIPTION

[0023] The technical solutions and effects of the present application are further described in detail below with reference to the embodiments and accompanying drawings. It should be understood that the specific embodiments described herein are merely for explaining the present invention, and are not intended to limit the present invention. It should also be noted that, for ease of description, the accompanying drawings only illustrate portions of the present invention, not all of its components.

[0024] Example 1: Model construction

[0025] Eighteen-month-old male C57BL / 6J mice, weighing an average of approximately 30-40 g, were used. Mice were anesthetized with isoflurane and placed in the supine position on a dedicated mouse operating table. The muzzle and mouth of the mouse were placed under an anesthesia mask with an isoflurane volume adjusted to 1.5-2.0%. After anesthesia, the abdomen was shaved, disinfected, and draped. The abdominal skin and peritoneum were incised, the intestines dissected, and the infrarenal abdominal aorta was exposed and isolated. Appropriately sized sterile gloves were used to isolate the infrarenal abdominal aorta from surrounding tissue. A sterile cotton pad soaked in 40 μL of 10 mg / mL elastase was then wrapped around the infrarenal abdominal aorta to temporarily close the abdominal cavity. After 40 minutes, the cotton pad and gloves were removed, and any residual fluid in the abdominal cavity was gently wiped with a sterile cotton swab moistened with normal saline. The peritoneum and skin were sutured separately with 5-0 surgical sutures. After the mice regained consciousness, they were transferred to their cages and continued feeding. After 14 days, the mice were sacrificed, and samples were collected for subsequent testing.

[0026] Example 2: Morphological evaluation of thoracic aortic aneurysms in mice

[0027] The cross-sectional diameter of the ascending aorta in mice was measured using a high-resolution small animal ultrasound imaging system. Mice were anesthetized with isoflurane and fixed in the supine position on a dedicated mouse operating table. The snout and mouth of the mouse were placed in an anesthesia mask, and the isoflurane dosage was adjusted to 1.5% to 2.0%. Transthoracic echocardiography was performed while the mice were fixed in the supine position. Ultrasound was used to examine the elasticity and cross-sectional dimensions of the mouse aorta, and clear, stable ultrasound images and videos were obtained. After the mice were sacrificed, the heart and aorta were isolated, and the maximum external diameter of the ascending aorta was measured and photographed. Partial tissue sections were then sectioned and stained with hematoxylin and eosin, Masson's trichrome staining, and the internal diameter of the aortic aneurysm at its maximum external diameter was measured under a microscope.

[0028] Figure 1 The pathological differences between young mice (6-8 weeks) and old mice (18 months) in the PPE-induced mouse abdominal aortic aneurysm model were demonstrated. Through HE and EVG staining, this study observed the histological changes of the aortic wall in detail (Figures A and C). HE staining results showed that after PPE treatment, the aortic media of the mice in the Old-PPE group was significantly thickened (62.27±5.0μm), and the structural changes of the media were more significant compared with the Young-PPE group (43.31±1.3μm) (Figure B). Specifically, there was a reduction in smooth muscle cells, rearrangement of collagen fibers, and more severe damage to elastic fibers. These changes suggest that the structural integrity and mechanical properties of the aortic wall of aged mice are more significantly damaged.

[0029] EVG staining further confirmed that the Old-PPE group showed more severe elastic layer disruption and elastic fiber loss, indicating a significant decrease in the elasticity and tensile strength of the aorta in aged mice (Figure C). This result suggests that the stress response and repair capacity of the aortic wall in aged mice are significantly weakened compared to young mice.

[0030] Ultrasound evaluation results showed that at one and two weeks after PPE treatment, the maximum diameter of the dilated abdominal aorta in aged mice [(1.32±0.19mm)1W and (1.26±0.10mm)2W] was significantly higher than that in young mice [(0.95±0.01mm)1W and (1.14±0.14mm)2W]. These results indicate that the differences in the rate and severity of aortic aneurysm development in the aged mouse model are more consistent with our clinical findings that abdominal aortic aneurysms are more common in the elderly.

[0031] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed, but as long as they are within the scope of the claims of the present application, they are protected by the patent law.

Claims

1. A method for constructing a mouse abdominal aortic aneurysm model, characterized in that: The method involves inducing abdominal aortic aneurysm formation in 18-month-old mice by wrapping the infrarenal abdominal aorta with a sterile cotton pad soaked with elastase.

2. The method for constructing a mouse abdominal aortic aneurysm model according to claim 1, wherein: The 18-month-old mice are 18-month-old male C57BL / 6J mice with an average body weight of 30-40 g.

3. The method for constructing a mouse abdominal aortic aneurysm model according to claim 1, wherein: The sterile cotton piece is soaked with 35-45 uL of 10 mg / mL elastase.

4. The method for constructing a mouse abdominal aortic aneurysm model according to claim 1, wherein: The infrarenal abdominal aorta is wrapped with a sterile cotton pad soaked in elastase for 30 to 50 minutes.

5. The method for constructing a mouse abdominal aortic aneurysm model according to claim 1, wherein: The construction method comprises the following steps: using 18-month-old male C57BL / 6J mice with an average weight of 30-40 g, anesthetizing them with isoflurane, fixing the mice in a supine position, placing the nose and mouth of the mice in an anesthesia mask, and adjusting the amount of isoflurane to 1.5%-2.0%; after anesthesia, removing the abdominal hair, disinfecting, and laying a drape; cutting the abdominal skin and peritoneum, stripping the intestines, exposing and separating the infrarenal abdominal aorta, using sterile gloves of appropriate size to isolate the infrarenal abdominal aorta from surrounding tissues, and then wrapping the infrarenal abdominal aorta with a sterile cotton pad soaked in 40 μL, 10 mg / mL elastase to temporarily close the abdominal cavity; removing the cotton pad and gloves after 40 minutes, gently wiping away the residual fluid in the abdominal cavity with a sterile cotton swab dipped in physiological saline, suturing the peritoneum and skin with 5-0 surgical sutures, and transferring the mice to a cage after they regain consciousness and continuing to be fed.

6. A mouse abdominal aortic aneurysm model obtained by the construction method according to any one of claims 1 to 5.

7. Use of the mouse model constructed by the method for constructing a mouse abdominal aortic aneurysm model according to any one of claims 1 to 5 in screening drugs for treating and / or preventing aging-related aneurysm diseases.

8. The use according to claim 7, characterized in that The aging-related aneurysmal diseases include abdominal aortic aneurysms in elderly patients.

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