A rat model of acute pulmonary thromboembolism and a method for constructing the same

Abstract

The application relates to the technical field of medicines, and particularly discloses an acute pulmonary thromboembolism rat model and a construction method thereof. The application successfully induces pathological changes such as pulmonary artery thrombosis by a method of inducing deep vein thrombosis, falling off and blocking the pulmonary artery, and the acute pulmonary thromboembolism animal model has good stability and high reproducibility, can be used for the research on pathophysiological mechanisms of acute pulmonary thromboembolism, the prediction of disease occurrence, the research and evaluation of treatment strategies, and thus can be more beneficial to the treatment of acute pulmonary thromboembolism patients, promote good prognosis, explore new targets and develop new drugs, so as to reduce the mortality of acute pulmonary thromboembolism patients.

Description

Technical Field

[0001] This invention relates to the field of medical technology, and in particular to an animal model of acute pulmonary thromboembolism. Background Technology

[0002] Venous thromboembolism includes deep vein thrombosis (DVT) and acute pulmonary thromboembolism (APVT). AVT is clinically common and can be fatal. The mortality and disability rates of AVT are both high. DVT has a high incidence rate globally, and AVT primarily results from the detachment of DVT thrombi. High-risk AVT often leads to sudden death. The 7-day all-cause mortality rate for AVT is 1.9%-2.9%, and the 30-day all-cause mortality rate is 4.9%-6.6%. A serious complication following AVT, chronic thromboembolic pulmonary hypertension, occurs in 0.1%-9% of cases. Therefore, AVT has a poor prognosis and severely impacts patients' quality of life. Acute pulmonary thromboembolism (APTE) is currently difficult to treat, has a high mortality rate, and lacks effective clinical methods for prediction and intervention. The main treatment includes anticoagulation, but its efficacy and prognosis are poor. Other treatments such as thrombolysis, interventional procedures, and surgical thrombectomy carry extremely high risks and have poor outcomes. This is related to the fact that the pathophysiological mechanisms of APT remain unclear. However, animal models of APT are of great value in exploring its pathogenesis, pathophysiological processes, predicting disease occurrence, and determining treatment strategies.

[0003] Animal models of acute pulmonary thromboembolism can be used to simulate acute pulmonary thromboembolism in humans. By studying animal disease models, we can conduct more in-depth research and exploration into the etiology and pathophysiological process of acute pulmonary thromboembolism, which is conducive to the development of new disease prediction methods and therapeutic targets, and promotes a good disease outcome. Therefore, the scientific research value of animal models of acute pulmonary thromboembolism is enormous.

[0004] Current animal models of acute pulmonary thromboembolism (APTE) in pigs, dogs, rabbits, and rats cannot simulate the pathophysiological process in humans, from deep vein thrombosis to pulmonary artery blockage leading to APT. The APT rat model and its construction technology described in this patent application are not reported domestically or internationally. However, clinical practice has shown that APT has a high morbidity and mortality rate; therefore, there is an urgent need for an animal model that can simulate APT. Summary of the Invention

[0005] To address the lack of existing animal models and their construction techniques, the present invention aims to provide a rat model of acute pulmonary thromboembolism and a method for constructing it. To achieve this objective, the technical solution of the present invention is as follows: In a first aspect, the present invention provides a method for preparing a rat model of acute pulmonary thromboembolism, comprising the following steps: (1) After fasting for 12 hours before surgery, the rats were anesthetized by intraperitoneal injection of ready-to-use tribromoethanol, fixed on a rat board, and the hair on the midline of the rat's abdomen was trimmed. After disinfection with iodine, an incision of about 2 cm was made in the midline of the abdomen under aseptic conditions to expose and separate the inferior vena cava branches below the left renal vein to the level of the iliac vein. (2) The inferior vena cava branches on both sides were ligated with 5-0 sutures. The abdominal aorta and inferior vena cava were separated about 0.5 cm below the junction of the left renal vein and the inferior vena cava. Another 4-0 suture was taken and placed parallel to the main trunk of the inferior vena cava. The 5-0 suture was passed through the inferior vena cava and ligated. The parallel 4-0 sutures were removed. The peritoneum and abdominal muscles were sutured using the continuous method, and the outer skin was sutured using the breakpoint method. The incision was wiped with iodine. After the rats recovered, they were able to eat and drink freely. (3) One day later, the abdomen was opened along the original incision, and the inferior vena cava that was subtotal ligated was found. The formation of thrombus in the inferior vena cava was observed. The thrombus was squeezed with a hemostat. The thrombus in the blood vessel was broken and flowed along the blood circulation of the inferior vena cava to embolize the pulmonary artery. After observing that there was no thrombus in the inferior vena cava, the squeezing was stopped. The peritoneum and abdominal muscles were sutured using the continuous method, and the outer skin was sutured using the breakpoint method. The incision was wiped with iodine. After the rat recovered, it was able to drink and drink freely.

[0006] Secondly, the present invention provides an animal model of acute pulmonary thromboembolism, which is constructed by the aforementioned preparation method.

[0007] The animal model described herein exhibits the progression from deep vein thrombosis to pulmonary embolism, resulting in pulmonary thromboembolism, thus mirroring the pathophysiological changes of acute pulmonary thromboembolism in humans. The animal model provided by this invention can simulate acute pulmonary thromboembolism in humans and can be used as an animal model for the research, prediction, and evaluation of treatment strategies for acute pulmonary thromboembolism, demonstrating promising application prospects.

[0008] Thirdly, the present invention further provides the application of the animal model in predicting the occurrence of acute pulmonary thromboembolism and in studying or screening therapeutic targets or drugs for acute pulmonary thromboembolism, as well as the application of the animal model in evaluating the therapeutic effect of acute pulmonary thromboembolism.

[0009] The animal model provided by this invention can be used to study and explore the pathophysiological mechanism of acute pulmonary thromboembolism in humans, predict the occurrence of the disease, and study and explore new therapeutic targets. It can also be used for the development of new drugs for the treatment of acute pulmonary thromboembolism, which may be beneficial to the treatment and prognosis of patients with acute pulmonary thromboembolism.

[0010] The raw materials or reagents involved in this invention are all commercially available products, and the operations involved are conventional operations in the field unless otherwise specified. The beneficial effects of this invention are as follows: This invention constructs an animal model of acute pulmonary thromboembolism by inducing deep vein thrombosis (DVT) formation, detachment, and pulmonary artery blockage. This method can induce pathophysiological changes such as DVT detachment, pulmonary artery blockage, and pulmonary artery thrombosis formation. The animal model is stable, durable, and reproducible, and can be used to study the pathophysiological mechanisms of acute pulmonary thromboembolism, predict disease occurrence, and research and evaluate treatment strategies. This may be more beneficial for the treatment of patients with acute pulmonary thromboembolism, promote a better prognosis, explore new targets, and develop new drugs to reduce the mortality rate of patients with acute pulmonary thromboembolism.

[0011] The method for constructing the animal model of this invention is simple to operate and requires inexpensive materials. The animal model exhibits good stability, with high stability and reproducibility of pathological changes such as pulmonary thrombosis, and can effectively simulate acute pulmonary thromboembolism in humans. Attached Figure Description

[0012] The accompanying drawings are incorporated in and form part of this specification.

[0013] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0014] Figure 1. Subtotal ligation of the inferior vena cava in rats; Figure 2. Arrows indicate inferior vena cava thrombosis in rats; Figure 3. Arrows indicate pulmonary artery thrombosis in rats; Figure 4 HE staining image, arrows indicate rat pulmonary artery thrombosis. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings.

[0016] Unless otherwise specified, the experimental methods used in the following steps are standard methods. Unless otherwise specified, all materials and reagents used in the following steps are commercially available.

[0017] An acute pulmonary thromboembolism rat model and its construction method, specifically including the following steps: Step 1: Inducing deep vein thrombosis (e.g.) Figure 1 , 2 ): After fasting for 12 hours preoperatively, rats were anesthetized with ready-to-use tribromoethanol via intraperitoneal injection. They were then fixed to a rat board, and the hair on the midline of the abdomen was trimmed. After disinfection with povidone-iodine, a 2cm incision was made under aseptic conditions in the midline of the abdomen to expose and separate the inferior vena cava branches below the left renal vein to the level of the iliac vein. The inferior vena cava branches on both sides were ligated with 5-0 sutures. The abdominal aorta and inferior vena cava were separated approximately 0.5cm below the junction of the left renal vein and inferior vena cava. A 4-0 suture was placed parallel to the main trunk of the inferior vena cava, and the 5-0 suture was passed through the inferior vena cava and ligated. The parallel 4-0 sutures were then removed. The peritoneum and abdominal muscles were sutured using a continuous suture method, and the outer skin was sutured using a breakpoint suture method. The incision site was wiped with povidone-iodine. After recovery, the rats were able to drink and drink freely.

[0018] Step Two: Pulmonary Artery Thrombosis ( Figure 3 ): One day later, the abdomen was reopened along the original incision to locate the subtotal ligated inferior vena cava. Thrombosis of the inferior vena cava was observed. The thrombus was squeezed with vascular clamps, and the thrombus inside the blood vessel fragmented and flowed along the blood circulation of the inferior vena cava to embolize the pulmonary artery. After observing that there was no thrombus in the inferior vena cava, the squeezing was stopped. The peritoneum and abdominal muscles were sutured using the continuous method, and the outer skin was sutured using the breakpoint method. The incision site was wiped with iodine. After the rat recovered, it was able to eat and drink freely.

[0019] Step 4: Obtain rat lung tissue specimens to verify successful model establishment (e.g., Figure 4 ): Three days later, the rats were euthanized by exsanguination, and their lung tissue was collected for observation. Half of the lung tissue was frozen at -80℃, while the other half was stained with hematoxylin and eosin (HE). Rats used in the model were randomly sampled to establish an acute pulmonary thromboembolism model. The sampled rats showed stable symptoms, and HE staining results showed pulmonary artery thrombosis as indicated by the arrow, demonstrating the feasibility and stability of the above method for preparing the animal model. Finally, the rat carcasses were disposed of harmlessly.

[0020] This invention confirms the feasibility, stability, and reproducibility of animal models of acute pulmonary thromboembolism (APTE). By establishing an APTE model that is simple to operate, economical to use, has a high success rate, and is durable and stable, it provides sufficient laboratory evidence for clinically relevant APTE research. It can be applied to the prediction, research, or screening of APTE therapeutic targets or drugs, as well as to the evaluation of APTE treatment efficacy.

[0021] The specific embodiments described above are further detailed explanations of the purpose, technical solutions, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A rat model of acute pulmonary thromboembolism and its construction method, characterized in that, Includes the following steps: (1) Using rats as the animal model, after anesthesia, a 2cm incision was made in the middle of the abdomen to expose and separate the inferior vena cava branches below the left renal vein to the level of the iliac vein. The inferior vena cava branches on both sides were ligated with 5-0 sutures. The abdominal aorta and inferior vena cava were separated about 0.5cm below the junction of the left renal vein and the inferior vena cava. Another 4-0 suture was taken and placed parallel to the main trunk of the inferior vena cava. The 5-0 suture was passed through the inferior vena cava and ligated. The parallel 4-0 sutures were removed. The peritoneum and abdominal muscles were sutured using the continuous method, and the outer skin was sutured using the breakpoint method. The incision site was wiped with iodine. After the rats recovered, they were able to drink and drink freely. (2) One day later, the abdomen was opened along the original incision, and the inferior vena cava that was subtotal ligated was found. The formation of thrombus in the inferior vena cava was observed. The thrombus was squeezed with a hemostat. The thrombus in the blood vessel was broken and flowed along the blood circulation of the inferior vena cava to embolize the pulmonary artery. After observing that there was no thrombus in the inferior vena cava, the squeezing was stopped. The peritoneum and abdominal muscles were sutured using the continuous method, and the outer skin was sutured using the breakpoint method. The incision was wiped with iodine. After the rat recovered, it was able to drink and drink freely.

2. The application of the animal model constructed by the preparation method of claim 1 in predicting the occurrence of acute pulmonary thromboembolism and in studying or screening therapeutic targets or drugs for acute pulmonary thromboembolism.

3. The application of the animal model constructed by the preparation method of claim 1 in evaluating the therapeutic effect of acute pulmonary thromboembolism.

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