Hypertension mouse model and construction method and application thereof

Abstract

The invention discloses a hypertension mouse model. The hypertension mouse model is obtained by intragastric administration of hypoxanthine and oteracil potassium salt into a mouse for a period of time. The invention also discloses a construction method and application of the gene. The invention provides a novel hypertension modeling mode, hypoxanthine and oteracil potassium salt are combined for use, and the administration dosage and administration mode of hypoxanthine and oteracil potassium salt are controlled, so that the serum uric acid level of a model mouse is stably increased, the blood pressure of the model mouse is increased stably and lastingly, the direct damage to liver and kidney functions is small, and the model mouse has a good application prospect. Therefore, the uric acid induced hypertension mouse model is successfully constructed. The uric acid induced hypertension mouse model provides a valuable tool for researching the pathogenesis of hypertension, evaluating a new treatment method and understanding the effect of serum hyperuricemia in hypertension.

Description

Technical Field

[0001] The present invention relates to the technical field of animal models, and in particular to a hypertensive mouse model and a construction method and application thereof. Background Art

[0002] Hypertension animal models are important tools for studying the pathogenesis of the disease and developing treatments. The following are some existing hypertension animal model technologies and their shortcomings:

[0003] 1. Spontaneously hypertensive rat (SHR) or spontaneously hypertensive mouse (such as BPH / 2J) model: It has certain requirements for feeding conditions and a long breeding cycle. The price is slightly higher than that of ordinary SD rats. Genetic breeding is troublesome, and it is easy to mutate and discontinue. The complications are complex and it is difficult to use them in large quantities.

[0004] 2. Dahl salt-sensitive hypertensive rat model: It has limitations such as shortened lifespan, certain requirements for feed and high price.

[0005] 3. Surgical hypertension animal model: It causes greater damage to the kidneys, but the degree of blood pressure increase is limited. It requires dietary intervention and is not suitable for studying its functional and structural damage.

[0006] 4. Animal models of drug-induced hypertension: Some are difficult to maintain stable and long-term high pressure, some are complicated to operate and costly, and some are prone to drug overdose, leading to weight loss in animals, and in severe cases, death of animals.

[0007] 5. Diet / environment-induced hypertension animal model: This type of method takes a long time to establish a model, has a low success rate, and requires strict control of diet / environment, which to some extent limits the research and application of this type of model.

[0008] In summary, although a variety of animal models of hypertension have been established, each model has its limitations, and currently most models of hypertension are concentrated in rat models. Therefore, we developed a hypertensive mouse model that has little impact on organs such as the liver and kidneys, is simple to operate, and has low cost. Summary of the invention

[0009] The purpose of the present invention is to provide a hypertensive mouse model and a construction method and application thereof, so as to solve the deficiencies of the prior art.

[0010] The present invention adopts the following technical solutions:

[0011] The first aspect of the present invention provides a hypertensive mouse model, wherein the hypertensive mouse model is constructed by the following method:

[0012] The mice were orally gavaged with hypoxanthine and potassium salt of oxalic acid for a period of time to obtain the hypertensive mouse model.

[0013] Furthermore, the dosage of hypoxanthine was 1200 mg / kg mouse / d, and the dosage of potassium salt of oxonate was 1200 mg / kg mouse / d, and they were continuously gavaged for 8 weeks.

[0014] Furthermore, the concentration of hypoxanthine is 48 mg / ml, and the concentration of oxonic acid potassium salt is 48 mg / ml.

[0015] Furthermore, the mice are 6-10 weeks old.

[0016] The second aspect of the present invention provides a method for constructing a hypertensive mouse model, comprising the following steps:

[0017] The mice were orally gavaged with hypoxanthine and potassium salt of oxalic acid for a period of time to obtain the hypertensive mouse model.

[0018] Furthermore, the dosage of hypoxanthine was 1200 mg / kg mouse / d, and the dosage of potassium salt of oxonate was 1200 mg / kg mouse / d, and they were continuously gavaged for 8 weeks.

[0019] Furthermore, the concentration of hypoxanthine is 48 mg / ml, and the concentration of oxonic acid potassium salt is 48 mg / ml.

[0020] Furthermore, the mice are 6-10 weeks old.

[0021] The third aspect of the present invention provides the use of the above-mentioned hypertensive mouse model in studying the pathogenesis of hypertension.

[0022] The fourth aspect of the present invention provides the use of the above-mentioned hypertensive mouse model in the screening of drugs for preventing and treating hypertension and the evaluation of their efficacy.

[0023] Beneficial effects of the present invention:

[0024] The present invention provides a novel method for modeling hypertension, which combines hypoxanthine and oxonic acid potassium salt and controls the dosage and administration method of hypoxanthine + oxonic acid potassium salt, so that the serum uric acid level of the model mice is stably increased, the model mice have increased blood pressure, which is stable and lasting, and the direct damage to liver and kidney functions is small, thereby successfully constructing a uric acid-induced hypertension mouse model.

[0025] The uric acid-induced hypertension mouse model of the present invention has the following advantages:

[0026] 1. Simulate the pathological characteristics of human hypertension: The uric acid-induced hypertensive mouse model of the present invention has a stable and persistent phenotype of elevated blood pressure and can be used for hypertension-related research.

[0027] 2. Low risk of direct damage to the liver and kidneys: The uric acid-induced hypertension mouse model of the present invention causes little direct damage to liver and kidney function and only causes increased blood pressure, thus successfully constructing a hypertension model without nonspecific organ damage.

[0028] 3. Simple operation and low cost: In the construction of the uric acid-induced hypertension mouse model of the present invention, the administration method of hypoxanthine + potassium salt of oxalic acid is oral gavage. The construction method is simple to operate, low in cost, and easy to repeat experiments and large-scale application.

[0029] 4. Provides a stable model: Hypoxanthine is a product of purine metabolism in the body and is converted into uric acid under the action of xanthine oxidase. Oxononic acid potassium salt is a commonly used uricase inhibitor, which blocks the degradation of uric acid by inhibiting the activity of uricase, thereby leading to increased serum uric acid levels. The present invention combines the two to increase uric acid production while reducing uric acid excretion, thereby creating a high uric acid level in the body. This type of modeling method under the influence of multiple factors is more stable than a single drug administration modeling method, and the increase in uric acid levels is more significant.

[0030] 5. Better understanding of the pathogenesis of hypertension: The uric acid-induced hypertensive mouse model of the present invention is of great value in exploring the occurrence and development mechanism of hypertension, especially in the study of hypertension related to high uric acid levels, its advantages are particularly significant.

[0031] In summary, the uric acid-induced hypertensive mouse model of the present invention provides a valuable tool for studying the pathogenesis of hypertension, evaluating new treatments, and understanding the role of serum high uric acid in hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 A is a line graph of blood pressure changes in the control group and the experimental group. A is a line graph of blood pressure changes in the control group and the experimental group of female mice; B is a line graph of blood pressure changes in the control group and the experimental group of male mice.

[0033] Figure 2 A is a statistical graph of the urine protein / urine creatinine ratio of the control group and the experimental group. B is a statistical graph of the urine protein / urine creatinine ratio of the control group and the experimental group of male mice.

[0034] Figure 3 A is a statistical graph of serum alanine aminotransferase levels in the control group and the experimental group. A is a statistical graph of serum alanine aminotransferase levels in the control group and the experimental group of female mice; B is a statistical graph of serum alanine aminotransferase levels in the control group and the experimental group of male mice.

[0035] Figure 4A is a statistical graph of serum uric acid levels in the control group and the experimental group. A is a statistical graph of serum uric acid levels in the control group and the experimental group of female mice; B is a statistical graph of serum uric acid levels in the control group and the experimental group of male mice.

[0036] Figure 5 A is a statistical graph of serum creatinine levels in the control group and the experimental group. A is a statistical graph of serum creatinine levels in the control group and the experimental group of female mice; B is a statistical graph of serum creatinine levels in the control group and the experimental group of male mice.

[0037] Figure 6 A is a statistical graph of serum total cholesterol levels in the control group and the experimental group. A is a statistical graph of serum total cholesterol levels in the control group and the experimental group of female mice; B is a statistical graph of serum total cholesterol levels in the control group and the experimental group of male mice.

[0038] Figure 7 A is a statistical graph of serum triglyceride levels in the control group and the experimental group. A is a statistical graph of serum triglyceride levels in the control group and the experimental group of female mice; B is a statistical graph of serum triglyceride levels in the control group and the experimental group of male mice.

[0039] Figure 8 A is a statistical graph of serum blood glucose levels in the control group and the experimental group. A is a statistical graph of serum blood glucose levels in the control group and the experimental group of female mice; B is a statistical graph of serum blood glucose levels in the control group and the experimental group of male mice.

[0040] In the above figures, * indicates P ≤ 0.05. DETAILED DESCRIPTION

[0041] The present invention is further explained below in conjunction with the examples and drawings. The following examples are only used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0042] Example 1

[0043] 1.1 Construction of uric acid-induced hypertensive mouse model

[0044] 32 8-10 week old C57BL / 6 mice (weight 20-24g) were ordered, including 16 female mice and 16 male mice, and adaptively raised for one week. They were divided into female control group, female experimental group (female uric acid-induced hypertension group), male control group, and male experimental group (male uric acid-induced hypertension group), with 8 mice in each group. Each mouse in the control group was orally gavaged with 1X PBS (diluted with 20XPBS (Sangon Biotech, B548117-0500), pH 7.2-7.6) 500ul per day; the experimental group was orally gavaged with hypoxanthine (ABCONE, H03235) 1200mg / kg mouse + oxonate potassium salt (COOLABER, CO8101) 1200mg / kg mouse per day, hypoxanthine and oxonate potassium salt were prepared with 1X PBS (same as above), and the prepared concentration was 48mg / ml; continuous gavage for 8 weeks (maintaining gavage at the same time every day). Adaptive feeding was carried out in an SPF environment, with normal feeding of feed and water, and subsequent experiments were carried out in a clean environment, with normal feeding of feed and water.

[0045] 1.2 Pathophysiological characteristics of the uric acid-induced hypertensive mouse model

[0046] (1) Before starting the gavage operation (i.e., before gavage after one week of adaptive feeding), measure blood pressure (systolic pressure, the same below) twice (the average of the two adaptive blood pressure measurements is recorded as the blood pressure at week 0) to prevent changes in blood pressure caused by stress. Figure 1 The blood pressure was measured in a week (the average blood pressure of the experiment was used as the statistical data). The blood pressure level was closely monitored until the end of gavage. All blood pressure measurements were performed before gavage to prevent the mice from being affected by the operation. When measuring blood pressure, two pre-measurements were made (the purpose was to allow the mice to be measured in a quiet state to see whether the measurement results were stable and whether the instrument detected the blood pressure. After confirmation, the subsequent formal measurement was performed), and 10 formal measurements were performed. At least 6 valid measurements were made in the formal measurement, and the average value was taken.

[0047] (2) After intragastric administration, the mice were placed in metabolic cages and their urine was collected for 24 h to detect urine protein and urine creatinine.

[0048] (3) After collecting urine for 24 hours, the mouse eyeballs were removed and blood was collected. Serum was collected to test alanine aminotransferase, uric acid, creatinine, total cholesterol, triglycerides, and blood glucose. The mice were euthanized after the experiment.

[0049] Blood pressure was measured with a sphygmomanometer, and 24-hour urine and serum tests were performed with a biochemical analyzer.

[0050] The changes of blood pressure in each group Figure 1As shown, the results showed that the blood pressure of the uric acid-induced hypertension group began to rise in the third week, and then its blood pressure level showed a steady increase, showing a hypertensive phenotype.

[0051] The results of urine protein and urine creatinine in each group are as follows Figure 2 The results of serum tests in each group are shown in Figure 3-8 The results showed that there was no significant change in serum alanine aminotransferase, creatinine, total cholesterol and blood sugar in the uric acid-induced hypertension group. The uric acid-induced hypertension of the present invention has less direct damage to liver and kidney function.

Claims

1. A hypertensive mouse model, characterized in that: The hypertensive mouse model was constructed by the following method: The mice were orally gavaged with hypoxanthine and potassium salt of oxalic acid for a period of time to obtain the hypertensive mouse model.

2. A hypertensive mouse model according to claim 1, characterized in that: The dosage of hypoxanthine was 1200 mg / kg mouse / d, and the dosage of potassium salt of oxonate was 1200 mg / kg mouse / d, and they were gavaged continuously for 8 weeks.

3. A hypertensive mouse model according to claim 2, characterized in that: The concentration of hypoxanthine is 48 mg / ml, and the concentration of potassium salt of oxonate is 48 mg / ml.

4. A hypertensive mouse model according to claim 1, 2 or 3, characterized in that: The mice are 6-10 weeks old.

5. A method for constructing a hypertensive mouse model, characterized in that: The steps include: The mice were orally gavaged with hypoxanthine and potassium salt of oxalic acid for a period of time to obtain the hypertensive mouse model.

6. The method for constructing a hypertensive mouse model according to claim 5, characterized in that: The dosage of hypoxanthine was 1200 mg / kg mouse / d, and the dosage of potassium salt of oxonate was 1200 mg / kg mouse / d, and they were gavaged continuously for 8 weeks.

7. The method for constructing a hypertensive mouse model according to claim 6, characterized in that: The concentration of hypoxanthine is 48 mg / ml, and the concentration of potassium salt of oxonate is 48 mg / ml.

8. The method for constructing a hypertensive mouse model according to claim 5, 6 or 7, characterized in that: The mice are 6-10 weeks old.

9. Use of the hypertensive mouse model according to any one of claims 1 to 4 in studying the pathogenesis of hypertension.

10. Use of the hypertensive mouse model according to any one of claims 1 to 4 in screening and evaluating the efficacy of drugs for preventing and treating hypertension.

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