A method for constructing a hyperlipidemia-resistant rat model

Abstract

The application discloses a method for constructing a hyperlipidemia-resistant rat model, and belongs to the technical field of experimental animal model construction. S1, basic population screening: abnormal lipid metabolism rats are selected, and the rats are fed with high-fat feed at the age of 4 weeks, and the feeding is continuously performed for 4 weeks; S2, blood lipid detection and preliminary screening: the animals are fasted for 12h, blood is taken from the submandibular vein, serum is separated by centrifugation at 3000r / min for 10min, and the total cholesterol, triglyceride, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol contents are detected by using an automatic biochemical analyzer; S3, paired reproduction and subculture: 30 male rats and 30 female rats are selected, and the rats are paired for reproduction at a ratio of 1:1 to obtain F1 generation; S4, inbreeding and stabilization: the F2-F4 generation rats are repeatedly fed, detected, screened and paired for reproduction according to steps S1-S3, and the F4 generation hyperlipidemia-resistant rat model is obtained through inbreeding, and the technical effect of obtaining the hyperlipidemia-resistant rat model which is stable in phenotype and can be inherited and subcultured is achieved through the combination of hybridization screening and inbreeding.

Description

Technical Field

[0001] This invention belongs to the field of experimental animal model construction technology, and particularly relates to a method for constructing a hyperlipidemia-resistant rat model. Background Technology

[0002] Hyperlipidemia is a common clinical disease and an independent risk factor for cardiovascular diseases such as stroke, coronary heart disease, and sudden cardiac death. Animal models of hyperlipidemia are of great significance for studying the pathological mechanisms of hyperlipidemia and preventing cardiovascular and cerebrovascular diseases. Rats are commonly used experimental animals for lipid metabolism research due to their ease of breeding, low cost, clear genetic background, and convenient blood collection. Normal rats are sensitive to high-fat diets, and short-term feeding can induce elevated blood lipids, making them suitable for establishing routine hyperlipidemia models. However, the cholesterol clearance rate in rats is significantly higher than in humans, and the increase in blood lipids in ordinary rats after induction with a high-fat diet is limited, making it difficult to meet the research needs of hyperlipidemia resistance mechanisms and metabolic tolerance regulation.

[0003] Existing hyperlipidemic rat models mostly focus on the elevated blood lipid phenotype, lacking stable, genetically resistant strains. This hinders the systematic analysis of the molecular mechanisms by which the body defends against lipid metabolism disorders, limiting the discovery of new targets for the prevention and treatment of hyperlipidemia. Previous studies have found that Wistar rats with abnormal lipid metabolism, after being crossed with SD rats and selected through high-fat diets and inbreeding, can produce lipid-tolerant, lipid-resistant individuals. However, a standardized and reproducible method for model construction has not yet been established. Summary of the Invention

[0004] To address the problems existing in the prior art, this invention provides a method for constructing a hyperlipidemia-resistant rat model. This method combines hybridization screening with inbreeding to obtain a phenotypically stable and heritable hyperlipidemia-resistant rat model, thus solving the problems of limited hyperlipidemia elevation, lack of resistance phenotype, and difficulty in using the model for mechanistic research in the prior art.

[0005] This invention is achieved by a method for constructing a hyperlipidemia-resistant rat model, comprising the following steps: S1. Basic population screening: Selected rats with abnormal lipid metabolism and fed a high-fat diet starting at 4 weeks of age for 4 consecutive weeks. S2. Blood lipid detection and initial screening: Animals were fasted for 12 hours, blood was collected from the submandibular vein, and serum was separated by centrifugation at 3000 r / min for 10 min. The levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), and low-density lipoprotein cholesterol (LDL-c) were detected using a fully automated biochemical analyzer. S3. Pairing and breeding: 30 male and 30 female rats with TC significantly lower than ordinary SD rats in the same batch were selected and bred in a 1:1 ratio to obtain the F1 generation. S4. Inbreeding and Stabilization: The feeding, testing, screening, and pairing breeding of F2-F4 generation rats were repeated in steps S1-S3. The F4 generation of hyperlipidemia resistant rat models were obtained through inbreeding.

[0006] As a preferred embodiment of the present invention, in step S1, the lipid metabolism abnormal rat is a hybrid offspring of Wistar rat and SD rat with lipid metabolism abnormality.

[0007] This setup expands the genetic base and improves reproductive performance through hybridization, providing a qualified starting population for subsequent inbreeding screening.

[0008] As a preferred embodiment of the present invention, in step S2, the screening basis is the blood lipid index after 4 weeks of feeding with a high-fat diet, with TC value as the core screening indicator.

[0009] This setting ensures that the screening criteria are uniform and repeatable, guaranteeing that the screening direction is consistent in each generation and improving the stability of the model's phenotype.

[0010] As a preferred embodiment of the present invention, the constructed hyperlipidemia-resistant rat model showed that, after 4 weeks of feeding with a high-fat diet, the serum TC and LDL-c levels were significantly lower than those of normal SD rats, and the TC level was not significantly different from that of the normal diet group.

[0011] This setup clarifies the core phenotypic features of the model, ensuring that the high-lipid resistance characteristics are stable and measurable, making it suitable for mechanism studies and drug evaluation.

[0012] An application of a hyperlipidemia-resistant rat model, applicable to the method for constructing the hyperlipidemia-resistant rat model, wherein the hyperlipidemia-resistant rat model is used in the study of the hyperlipidemia resistance mechanism.

[0013] This setup provides a stable and standardized animal tool for exploring the mechanisms of metabolic diseases, discovering targets, and screening new drugs.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention establishes a stable and heritable hyperlipidemia-resistant rat model for the first time through hybridization screening combined with inbreeding and passage, solving the problem of limited increase in blood lipids and lack of resistance phenotype in ordinary rats after hyperlipidemia induction. After feeding the model with a high-fat diet, the levels of TC and LDL-c are significantly lower than those of the control SD rats, and the TC level after feeding with a high-fat diet is not significantly different from that of the ordinary diet group, indicating a clear and stable hyperlipidemia-resistant phenotype. Furthermore, the model can be directly used to study the resistance mechanism of hyperlipidemia, screen the core factors of metabolic disorder tolerance, and evaluate the efficacy of lipid-lowering drugs, providing a new research tool for the prevention and treatment of hyperlipidemia and promoting the development of metabolic disease mechanisms and drugs. Attached Figure Description

[0015] Figure 1 This is a comparison chart of blood lipid levels between hyperlipidemia resistant rats constructed in this invention and control rats under different feed conditions. Detailed Implementation

[0016] To further understand the invention's content, features, and effects, the following embodiments are provided, and detailed descriptions are given in conjunction with the accompanying drawings.

[0017] The following is in conjunction with the appendix Figure 1 The structure of the present invention will be described in detail.

[0018] Example 1:

[0019] 1.1 Laboratory Animals and Feed The baseline population consisted of offspring from a cross between Wistar rats with abnormal lipid metabolism and SD rats, and 4-week-old SPF-grade rats. High-fat diet (HD) was used for model establishment, while normal diet (LFD) served as a control.

[0020] 1.2 Construction Steps S1. Basic population screening: Four-week-old hybrid rats with abnormal lipid metabolism were selected and randomly divided into a control group and a model group. The model group was fed a high-fat diet for 4 weeks, while the control group was fed a normal diet. S2. Blood lipid testing and initial screening: After feeding, fast for 12 hours, blood is collected from the submandibular vein, and serum is separated by centrifugation at 3000r / min for 10min. The levels of TC, TG, HDL-c and LDL-c are detected using a fully automated biochemical analyzer. S3. Pairing and breeding: Individuals with significantly lower TC values ​​than control SD rats in the model group were selected, and 30 males and 30 females were paired at a 1:1 ratio to obtain the F1 generation. S4. Inbreeding and Stabilization: The feeding, testing, screening, and pairing breeding of F2-F4 generation rats were repeated in steps S1-S3. The F4 generation of hyperlipidemia resistant rat models were obtained through inbreeding.

[0021] 1.3 Model Validation After 4 weeks of feeding with a high-fat diet, the serum TC and LDL-c levels in hyperlipidemia resistant rats were significantly lower than those in normal SD rats. The TC level in normal SD rats fed with a high-fat diet was significantly higher than that in the normal diet group, while the TC level in hyperlipidemia resistant rats fed with a high-fat diet was not significantly different from that in the normal diet group. The hyperlipidemia resistant phenotype was stable and could be stably passaged for subsequent studies.

[0022] This invention, through hybridization screening combined with inbreeding and passaging, establishes for the first time a stable and heritable rat model of hyperlipidemia resistance, addressing the issues of limited lipid elevation and lack of resistance phenotype in ordinary rats after hyperlipidemia induction. After being fed a high-fat diet, the model exhibits significantly lower levels of total cholesterol (TC) and LDL-c compared to the control SD rats, with no significant difference in TC levels between the high-fat diet group and the ordinary diet group, demonstrating a clear and stable hyperlipidemia resistance phenotype. This model can be directly used for research on hyperlipidemia resistance mechanisms, screening of core factors for metabolic disorder tolerance, and evaluation of the efficacy of lipid-lowering drugs, providing a novel research tool for the prevention and treatment of hyperlipidemia and advancing the development of metabolic disease mechanisms and drugs.

[0023] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0024] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for constructing a hyperlipidemia-resistant rat model, characterized in that, Includes the following steps: S1. Basic population screening: Selected rats with abnormal lipid metabolism and fed a high-fat diet starting at 4 weeks of age for 4 consecutive weeks. S2. Blood lipid detection and initial screening: Animals were fasted for 12 hours, blood was collected from the submandibular vein, and serum was separated by centrifugation at 3000 r / min for 10 min. The levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), and low-density lipoprotein cholesterol (LDL-c) were detected using a fully automated biochemical analyzer. S3. Pairing and breeding: 30 male and 30 female rats with TC significantly lower than ordinary SD rats in the same batch were selected and bred in a 1:1 ratio to obtain the F1 generation. S4. Inbreeding and Stabilization: The feeding, testing, screening, and pairing breeding of F2-F4 generation rats were repeated in steps S1-S3. The F4 generation of hyperlipidemia resistant rat models were obtained through inbreeding.

2. The method for constructing a hyperlipidemia-resistant rat model as described in claim 1, characterized in that: In step S1, the lipid metabolism abnormal rats are hybrid offspring of Wistar rats and SD rats with lipid metabolism abnormalities.

3. The method for constructing a hyperlipidemia-resistant rat model as described in claim 1, characterized in that: In step S2, the screening criteria are blood lipid levels after 4 weeks of feeding with a high-fat diet, with TC value as the core screening indicator.

4. The method for constructing a hyperlipidemia-resistant rat model as described in claim 1, characterized in that: The constructed hyperlipidemia-resistant rat model showed that, after 4 weeks of feeding with a high-fat diet, serum TC and LDL-c levels were significantly lower than those of normal SD rats, and the TC level was not significantly different from that of the normal diet group.

5. The application of a hyperlipidemia-resistant rat model, characterized in that, The method for constructing a hyperlipidemia-resistant rat model according to any one of claims 1-4 is applicable, and the hyperlipidemia-resistant rat model is used in the study of the hyperlipidemia resistance mechanism.

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