Hyperuricemia animal model and construction method thereof

Abstract

The invention belongs to the technical field of animal model construction, and particularly relates to a hyperuricemia animal model and a construction method thereof.The animal model can remarkably prolong the maintenance time of the model, and modeling through a method for constraining an experimental animal comprises the steps that A, constraint day constraining is conducted, specifically, the experimental animal is constrained for 3-6 hours through a constraining device, and water and food are forbidden in the constraining period; b, in a non-constraint period, normal diet and free water drinking are carried out; and C, the modeling time is at least 21 days. According to an existing modeling method, uric acid can recover to a normal level after drug withdrawal for 2-6 hours, and the maintenance time of the model is very short, however, the hyperuricemia animal model capable of remarkably prolonging the maintenance time of the model is effectively constructed, and after modeling is stopped for 3 days, the uric acid level in mouse plasma is still maintained at a relatively high stage. The construction method can also be matched with existing modeling methods such as high-purine diet and uricase inhibitors, high-pressure living environments and bad living and diet habits are considered, and the construction method conforms to the pathogenesis of hyperuricemia of modern people.

Description

Technical Field

[0001] The present invention belongs to the technical field of the construction of animal models, and particularly relates to a hyperuricemia animal model and a construction method thereof. Background Art

[0002] Hyperuricemia is an independent risk factor for diseases such as chronic kidney disease, hypertension, cardiovascular and cerebrovascular diseases, and diabetes, and is an independent predictor of premature death. It has become the second most concerned metabolic disease globally. A meta-analysis in 2020 showed that the total prevalence of hyperuricemia in China was greater than 10%. The Guidelines for the Diagnosis and Treatment of Hyperuricemia and Gout in China in 2021 also showed that the overall prevalence of hyperuricemia in China was as high as 13.3%. Therefore, it is particularly important to construct a hyperuricemia animal model that conforms to the pathogenesis of modern hyperuricemia for screening active ingredients for reducing uric acid.

[0003] Currently, the generally recognized methods for establishing hyperuricemia models mainly achieve high uric acid levels in mice by administering a high-purine diet, uricase inhibitors, or uric acid excretion inhibitors (uric acid transporter inhibitors). Commonly used hyperuricemia models are mainly established by a high-purine diet, or a combination of a high-purine diet with uricase inhibitors and uric acid transporter inhibitors, as shown in Table 1.

[0004] Table 1 Common methods for establishing hyperuricemia models

[0005]

[0006] For example, the patent with the publication number CN1698906 discloses the "Method for Replicating Acute or Persistent Hyperuricemia Model in Rats", with the publication date of November 23, 2005, and the applicant being Nanjing University of Traditional Chinese Medicine. This method involves using SD and Wistar healthy rats, and administering purine (hypoxanthine, xanthine) at a dose of 100 - 2000 mg / kg and uricase inhibitor (potassium oxonate) at a dose of 5 - 1000 mg / kg to the rats by gavage, subcutaneous injection, and intraperitoneal injection. Every 5 - 24 hours, the rats are medicated in the same way and with the same drug dosage for one to two weeks, and a stable acute or persistent hyperuricemia model in rats can be obtained. The patent with the publication number CN1698907 discloses the "Method for Replicating Acute Hyperuricemia Model in Mice", with the publication date of November 23, 2005, and the applicant being Nanjing University of Traditional Chinese Medicine. This patent involves using Kunming and ICR healthy mice, and administering hypoxanthine or xanthine at a dose of 100 - 1500 mg / kg and uricase inhibitor potassium oxonate at a dose of 10 - 1500 mg / kg to the mice by gavage, subcutaneous injection, or intraperitoneal injection. Experiments have shown that by using a uricase inhibitor in combination with purines, a stable acute hyperuricemia model in mice can be obtained within 1 hour. The patent with the publication number CN103977007A discloses the "Method for Constructing Acute Hyperuricemia Tree Shrew Model", with the publication date of August 13, 2014, and the applicant being Institute of Medical Biology, Chinese Academy of Medical Sciences. This patent involves intraperitoneally injecting a suspension of potassium oxonate into adult healthy tree shrews aged 1 - 3 years, with the dosage being 40 mg / kg - 100 mg / kg based on the body weight of the tree shrew, and an acute hyperuricemia tree shrew model can be obtained. The patent with the publication number CN104388467A discloses "A Method for Constructing a Spontaneous Hyperuricemia Model and Its Use", with the publication date of March 4, 2015, and the applicant being Li Changgui. This patent uses transcription activator-like effector nuclease to knockout the urate oxidase gene in the target mouse with gene knockout, and a spontaneous hyperuricemia mouse model can be obtained. The survival time of the mice is long, which is conducive to long-term experiments on such mice. The patent with the publication number 103860562A discloses "A Pharmaceutical Composition for Establishing a Hyperuricemia Animal Model and Its Application", with the publication date of June 18, 2014, and the applicant being Zeng Wen, etc. This patent uses ethambutol and uric acid to establish the model, and the model animals are rhesus monkeys. The modeling time is short, the model is stable, and the success rate is high, and a stable and reliable acute hyperuricemia model can be obtained. In addition, for uricase gene knockout mice, transcription activator-like effector nuclease technology is used to knockout the Uox gene, but this method has a low survival rate of mice and is expensive, and is not suitable for drug screening experiments.

[0007] Problems such as short model maintenance time (uric acid can return to normal levels 2 - 6 hours after drug withdrawal) and high incidence of liver and kidney damage in mice exist when using methods such as high - purine diet, uricase inhibitors, or uric acid excretion inhibitors. Especially the short model maintenance time of the hyperuricemia model makes it difficult to conduct pharmacological evaluations using this model.

[0008] In summary, the inventors of the present invention aim to provide a construction method that can significantly extend the maintenance time of the hyperuricemia animal model. Summary of the Invention

[0009] The technical problem to be solved by the present invention is to provide a hyperuricemia animal model that can significantly extend the model maintenance time.

[0010] The animals that can be used in the present invention for constructing the animal model include mammals and avian animals.

[0011] Further preferably, the mammals include primates, rodents, artiodactyls, canids, and felids;

[0012] Still further preferably, the primates are selected from monkeys and orangutans; the rodents are selected from mice, rats, rabbits, and guinea pigs; the artiodactyls are selected from donkeys, sheep, cows, horses, and pigs; the canids are selected from dogs, wolves, jackals, and foxes; the felids are selected from cats;

[0013] Further preferably, the avian animals are chickens, ducks, and geese;

[0014] According to economy, rats or mice are preferably used as experimental animals, and most preferably mice are used as experimental animals.

[0015] The method for constructing the animal model in the present invention uses the method of restraining the experimental animal to establish the model, including the following steps:

[0016] A. Restraint on the restraint day: The experimental animal is restrained for 3 - 6 hours using a restraint device, and water and food are prohibited during the restraint period;

[0017] B. Non - restraint period: Normal diet and free access to water;

[0018] C. The modeling time is at least 21 days.

[0019] In the above - mentioned technical solution:

[0020] The restraint in step A is continuous restraint or intermittent restraint.

[0021] Adopting continuous restraint in step A means that every day during modeling is a restraint day, and the experimental animal is restrained every day for 3 - 6 hours using a restraint device, and water and food are prohibited during the restraint period;

[0022] Step A uses intermittent restraint, which means that every other day during the modeling period is a restraint day. The experimental animals are restrained every other day using a restraint device for 3 - 6 hours, and food and water are withheld during the restraint period.

[0023] The restraint device described in Step A is a device that ensures the integrity of the limbs of the experimental animals, restricts the activities of the experimental animals but does not affect their breathing.

[0024] The restraint time described in Step A is 3 - 6 hours. This time selection mainly considers the impact of the restraint time on mice. If the restraint time is too long, it may lead to death due to dehydration and other conditions.

[0025] Preferably, the restraint time described in Step A is 6 hours.

[0026] The fasting and water - withholding time in Step B is equal to the restraint time.

[0027] The continuous restraint for the modeling time described in Step C is at least 21 days (calculated as three weeks).

[0028] The intermittent restraint for the modeling time described in Step C is at least 35 days (calculated as five weeks).

[0029] Preferably, the intermittent restraint for the modeling time described in Step C is at least 42 days (calculated as six weeks).

[0030] The construction of the animal model described in the present invention can also use the method of restraining experimental animals in combination with the existing method of constructing a hyperuricemia animal model, such as:

[0031] (1) Restraining experimental animals in combination with a high - purine diet for modeling, including the following steps:

[0032] A. Restraint on the restraint day: The experimental animals are restrained using a restraint device for 3 - 6 hours, and food and water are withheld during the restraint period;

[0033] B. Non - restraint period: The diet uses a mixed feed containing dry yeast powder or adenine, and water is provided ad libitum;

[0034] C. The modeling time is at least 21 days.

[0035] In Step B, the content of dry yeast powder in the mixed feed is 5 - 10%.

[0036] Preferably, in Step B, the content of dry yeast powder in the mixed feed is 10%.

[0037] In Step B, the content of adenine in the mixed feed is 0.05 - 0.1%.

[0038] Preferably, in Step B, the content of adenine in the mixed feed is 0.1%.

[0039] The daily feeding amount is calculated based on the body weight of the experimental animals, with adenine at 125 - 250 mg / kg and dry yeast powder at 12.5 - 25 g / kg.

[0040] Preferably, the daily feeding amount is calculated based on the body weight of the experimental animals, with adenine at 250 mg / kg and dry yeast powder at 25 g / kg.

[0041] (2) Restrain the experimental animals and cooperate with uricase inhibitors to establish a model, including the following steps:

[0042] A. Restraint on the restraint day: The experimental animals are restrained with a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period.

[0043] B. Non - restraint period: Normal diet, free access to water.

[0044] C. The modeling time is at least 21 days, and potassium oxonate is intraperitoneally injected in the last 1 - 2 weeks of modeling.

[0045] In step C, when intraperitoneally injecting potassium oxonate, the daily injection amount is calculated based on the body weight of the experimental animals, with potassium oxonate at 150 - 300 mg / kg.

[0046] Preferably, in step C, when intraperitoneally injecting potassium oxonate, the daily injection amount is calculated based on the body weight of the experimental animals, with potassium oxonate at 300 mg / kg.

[0047] (3) Restrain the experimental animals and cooperate with uric acid transporter inhibitors to establish a model, including the following steps:

[0048] A. Restraint on the restraint day: The experimental animals are restrained with a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period.

[0049] B. Non - restraint period: Normal diet, free access to water.

[0050] C. The modeling time is at least 21 days, and pyrazinamide is administered by gavage in the last 1 - 2 weeks of modeling.

[0051] In step C, when administering pyrazinamide by gavage, it is once a day, and the gavage amount is calculated based on the body weight of the experimental animals, with pyrazinamide at 200 - 400 mg / kg.

[0052] Preferably, when administering pyrazinamide by gavage, it is once a day, and the gavage amount is calculated based on the body weight of the experimental animals, with pyrazinamide at 400 mg / kg.

[0053] The present invention proposes a method for constructing a hyperuricemia animal model by chronic restraint stress. After stopping the drug administration of the existing modeling methods for 2 - 6 hours, the uric acid can return to the normal level, and the model maintenance time is very short. However, the present invention effectively constructs a hyperuricemia animal model with a significantly extended model maintenance time. Three days after stopping the modeling, the uric acid level in the mouse plasma still remains at a relatively high level. The construction method of the present invention also cooperates with the modeling methods of high - purine diet, uricase inhibitor, and uric acid transporter inhibitor, taking into account the high - pressure living environment and bad living and eating habits, which is in line with the pathogenesis of hyperuricemia in modern people and can also significantly extend the model maintenance time. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 Experimental protocol diagram.

[0055] Figure 2 Chronic restraint stress experimental device diagram.

[0056] Figure 3 Diagram of the drinking device for the sucrose preference test.

[0057] Figure 4 Diagram of the tail suspension test device. DETAILED DESCRIPTION OF THE INVENTION

[0058] The following will explain and illustrate the solution of the present invention in combination with embodiments. Those skilled in the art will understand that the following embodiments are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. For those not specified in the embodiments regarding specific technologies or conditions, they shall be carried out according to the technologies or conditions described in the literature in this field or according to the product specifications. For the reagents or instruments not specified by the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0059] The following describes the specific embodiments of the present invention but does not limit the present invention.

[0060] I. Experimental protocol: See Figure 1 .

[0061] II. Experimental process

[0062] 1. Continuous restraint modeling

[0063] After the C57BL / 6J mice are adaptively fed for 7 days, two behavioral experiments, namely the sucrose preference test and the tail suspension test, are used to detect whether the behavioral states of the mice are significantly different. The mice with similar behavioral states are screened out and randomly and evenly divided into a blank group and a restraint group. Then, chronic restraint stress modeling is carried out. After the modeling is completed, the above - mentioned behavioral detections are repeated again to judge whether stress occurs in the model group. Immediately after the behavioral detections, the samples are taken.

[0064] Among them, the self - made restraint device is shown in Figure 2, the restraint device is to use iron wire to burn multiple holes of appropriate size on the wall of a 50-ml centrifuge tube to keep the mouse breathing freely, and burn a small hole in the centrifuge tube lid so that the mouse's tail passes through the centrifuge tube to protect the mouse's tail from being crushed. The restraint device is selected to meet the standard of ensuring the integrity of the experimental animal's limbs, restricting the movement of the experimental animal but not affecting the breathing of the experimental animal, but not limited to forms such as Figure 2 the form of.

[0065] (1) Sucrose preference test: This test is divided into two parts: an adaptation training stage and a test stage. First, separate the mice into cages, one mouse per cage, number and weigh them. During the adaptation training stage, place two drinking devices made of 15-ml centrifuge tubes filled with the same ordinary drinking water in each cage for the first 3 days (as shown in Figure 3 ). After the adaptation period ends, replace the ordinary drinking water in one drinking device with a pre-prepared sucrose solution, and the other drinking device still contains ordinary drinking water. Record the weights of the ordinary drinking water and sucrose water in each tube respectively, and then place them in the positions where they were placed during the adaptation period. After 24 hours, exchange the positions of the drinking tubes to eliminate the mouse's preference for the drinking position. Record the amounts of ordinary drinking water and sucrose water consumed by each mouse in 48 hours (weigh the drinking tubes), and calculate the sucrose preference rate.

[0066] Formula for calculating the sucrose preference rate:

[0067] Sucrose preference rate = sucrose consumption (g) ÷ (sucrose consumption (g) + ordinary drinking water consumption (g)) × 100%.

[0068] (2) Tail suspension test: Stick the mouse's tail at the 1 / 3 position with tape and hang it upside down on the bracket so that the mouse's head is 15 cm away from the tabletop (as shown in Figure 4 ), time for 6 minutes, and count the time of remaining stationary in the last 4 minutes.

[0069] (3) Continuous restraint modeling experiment: After the sucrose preference test and the tail suspension test, group the mice and enter the chronic restraint stress modeling stage. During the experiment, after weighing the mice every day, the mice in the blank group are allowed to eat and drink freely without intervention, and the mice in the restraint group are respectively restrained in the experimental restraint device for 6 hours every day, and are prohibited from drinking and eating during the restraint period. The modeling time is 21 days.

[0070] After the establishment of the restraint stress model, the sucrose preference test and the tail suspension test were performed again to calculate the sucrose preference rate and the tail suspension immobility time. After the tail suspension test, the mice were sacrificed by eye enucleation for blood collection. The blood samples were allowed to stand at room temperature for 2 h, centrifuged at 3500 r / min for 10 min, and the supernatant was taken and stored at -20 °C for later use. Then, the mice were sacrificed by cervical dislocation, and their liver and kidney tissues were collected. An appropriate amount of liver tissue was cut, added with 9 times the weight of the liver homogenization medium, homogenized in a low-temperature environment using a homogenizer, and then centrifuged at 3500 r / min for 10 min. The supernatant was taken and stored at -20 °C for later use. The left kidney was fixed with 4% paraformaldehyde solution, and the right kidney and the remaining liver tissue were stored in an -80 °C refrigerator for later use.

[0071] 2. Intermittent restraint modeling

[0072] Except that the restraint interval was changed to 1 day, the other experimental methods were the same as those in "1. Continuous restraint modeling", and the modeling time was limited according to actual requirements.

[0073] 3. Continuous restraint (intermittent restraint) + high-purine diet modeling

[0074] The restraint modeling method in "1. Continuous restraint modeling" and the intermittent restraint modeling method were the same as those in "2. Intermittent restraint modeling". The high-purine diet plan was as follows: Dry yeast powder and adenine were evenly mixed into the feed so that the contents of dry yeast powder and adenine reached 10% and 0.1% respectively. The daily feeding amount was calculated according to the body weight of the experimental animals, with adenine at 250 mg / kg and dry yeast powder at 25 g / kg. During the modeling period, the mice were fed with a high-purine diet and allowed free access to water.

[0075] 4. Continuous restraint (intermittent restraint) + uricase inhibitor modeling

[0076] The restraint modeling method in "1. Continuous restraint modeling" and the intermittent restraint modeling method were the same as those in "2. Intermittent restraint modeling". The uricase inhibitor plan was as follows: Oxonic acid potassium is a uricase inhibitor and can be used to prepare a hyperuricemia model by inhibiting uricase activity. However, due to its strong adverse reactions such as peritoneal sclerosis and ascites after long-term administration. Therefore, oxonic acid potassium (300 mg / kg) was only given by intraperitoneal injection once a day in the last 2 weeks of modeling.

[0077] 5. Continuous restraint (intermittent restraint) + uric acid transporter inhibitor modeling

[0078] The restraint modeling method in "1. Continuous restraint modeling" and the intermittent restraint modeling method were the same as those in "2. Intermittent restraint modeling". The uric acid transporter inhibitor plan was as follows: Pyrazinamide is a uric acid transporter inhibitor and can be used to prepare a hyperuricemia model by inhibiting uric acid excretion. However, due to liver and kidney damage after long-term administration. Therefore, pyrazinamide (400 mg / kg) was only given by gavage once a day in the last 2 weeks of modeling.

[0079] III. Experimental Results

[0080] 1. Comparison between continuous restraint modeling and common hyperuricemia models

[0081] Continuous restraint group: The model was established according to the method of "1. Continuous restraint modeling" in "II. Experimental Methods".

[0082] High-purine diet group: High-purine diet and free drinking water. Dry yeast powder and adenine were evenly mixed into the feed so that the contents of dry yeast powder and adenine reached 10% and 0.1% respectively. The daily feeding amount was calculated according to the body weight of the experimental animals, with adenine at 250 mg / kg and dry yeast powder at 25 g / kg. During the modeling period, the mice were fed with high-purine feed and had free access to drinking water.

[0083] Uricase inhibitor + high-purine diet group: Normal diet and free drinking water. In the last two weeks of modeling, hypoxanthine at 500 mg / (kg·d) was injected subcutaneously once a day + potassium oxonate at 300 mg / (kg·d) was injected intraperitoneally once a day.

[0084] Urate transporter inhibitor + high-purine diet group: Normal diet and free drinking water. In the last two weeks of modeling, hypoxanthine at 500 mg / (kg·d) was injected subcutaneously once a day + pyrazinamide at 400 mg / (kg·d) was administered by gavage once a day.

[0085] Samples were taken 1 h, 2 h, and 3 d after the last administration. Blood was collected from the mouse's eyeballs. After the blood samples were left standing at room temperature for 2 h, they were centrifuged at 3500 r / min for 10 min, and the supernatant was taken and stored frozen at -20°C in the refrigerator. After the mice were sacrificed by cervical dislocation, their liver and kidney tissues were taken for preservation. An appropriate amount of liver tissue was cut, and homogenization medium 9 times the weight of the liver tissue was added. Homogenization was carried out using a homogenizer in a low-temperature environment. After centrifugation at 3500 r / min for 10 min, the supernatant was taken and stored frozen at -20°C in the refrigerator. The left kidney was fixed with 4% paraformaldehyde solution, and the other part and the remaining liver tissue were stored in a -80°C refrigerator.

[0086] This experiment first compared the effect of restraint (for 3 weeks) on uric acid levels and its comparison with common modeling methods. As shown in Table 2, in the high-purine diet group, uricase inhibitor + high-purine diet group, and urate transporter inhibitor + high-purine diet group, after continuous administration for 2 weeks, a hyperuricemia model could be maintained 1 h after the end of modeling. However, 3 days after the end of modeling, the serum uric acid levels of the mice all returned to normal values. In the restraint group, 3 days after the end of modeling, the serum uric acid level of the mice was still significantly higher than that of the blank group (p < 0.05). This result indicates that the chronic restraint stress of restraint modeling can not only lead to the occurrence of hyperuricemia in mice, but also significantly increase the serum uric acid level of mice, and the maintenance time of the hyperuricemia model is significantly longer than that of the common hyperuricemia modeling method group.

[0087] Table 2 Serum uric acid levels in mice under different modeling methods (μmol / L)

[0088]

[0089] *, Compared with the blank group, p < 0.05; **, Compared with the blank group, p < 0.01

[0090] 2. Application scenarios of the restraint and intermittent restraint modeling methods

[0091] Since hyperuricemia is a chronic metabolic disease, its treatment will also be a long-term process. When screening drugs for reducing uric acid, the situation of long-term medication will be faced. To solve the problem of long-term construction of hyperuric acid models, this experiment compared the differences between intermittent restraint modeling and continuous restraint modeling. The research results are shown in Table 3. When the hyperuric acid model needs to be maintained for more than six weeks (6w), continuous restraint modeling or intermittent modeling (restraint every other day) can be used. Compared with the continuous restraint modeling method, the experimental operation of the intermittent modeling method is simpler when preparing a long-term hyperuricemia model.

[0092] Table 3 Serum uric acid levels in mice under the restraint and intermittent restraint modeling methods (μmol / L)

[0093] Grouping After the model establishment (3d) Blank group 102±10.87 Continuous restraint group (model establishment time: 3w) 155.43±8.79 Intermittent restraint group (model establishment time: 3w) 104.06±7.97 Intermittent restraint group (model establishment time: 4w) 105.90±9.68 Intermittent restraint group (model establishment time: 5w) 138.07±16.72 Intermittent restraint group (model establishment time: 6w) <![CDATA[156.86±20.65 ** >

[0094] **, Compared with the blank group, p < 0.01

[0095] 3. Combined application of restraint (intermittent restraint) modeling and common hyperuric acid models

[0096] After verifying that restraint and intermittent restraint can construct a more stable and persistent hyperuricemia model, this experiment continued to observe the possibility of combining restraint and intermittent modeling with other modeling methods. The results are shown in Table 4. After combining the restraint modeling method (3w) and the intermittent restraint modeling method (6w) with high-purine diet, uricase inhibitor, and uric acid transporter inhibitor respectively, the hyperuric acid level in mice can still be maintained 3 days after the end of modeling. Therefore, this method is also applicable to combined use with other common modeling methods.

[0097] Table 4 Serum uric acid levels in mice after combined application of the restraint modeling method and other modeling methods (μmol / L)

[0098]

[0099] *, Compared with the blank group, p < 0.05

[0100] In summary, after stopping the drug administration of the existing modeling methods for 2 - 6 hours, the uric acid can return to the normal level, and the model maintenance time is very short. However, the present invention constructs a hyperuricemia animal model by means of chronic restraint stress, which can significantly extend the maintenance time of the hyperuricemia animal model. Three days after stopping the modeling, the uric acid level in the plasma of the mice still remains at a relatively high stage. The restraint method can be continuous daily restraint or intermittent restraint at regular intervals. When preparing a long - cycle hyperuricemia model, the experimental operation of the intermittent modeling method is simpler and easier to operate. The restraint (intermittent restraint) modeling construction method of the present invention also cooperates with the modeling methods of high - purine diet, uricase inhibitor, and uric acid transporter inhibitor, which can also significantly extend the model maintenance time. After continuous restraint, intermittent restraint is combined with other modeling methods, no obvious advantages are observed, which may be related to the fact that the modeling methods such as high - purine diet and uric acid metabolism enzyme inhibitor are rapidly metabolized three days after stopping the modeling. Thus, from the perspectives of economy and cost, only using continuous restraint and intermittent restraint without additional auxiliary modeling conditions can still successfully construct a hyperuric animal model, providing a new option for the construction method of animal models.

Claims

1. A method for constructing an animal model of hyperuricemia, characterized in that, Modeling is performed by the method of restraining experimental animals, including the following steps: A. Restraint on restraint days: The experimental animals are restrained by a restraint device, and water and food are prohibited during the restraint period. B. Non-restraint period: Normal diet, free access to water. C. The modeling time is at least 21 days.

2. The method for constructing a hyperuricemia animal model according to claim 1, wherein At least one of the following conditions is satisfied: The restraint described in step A is continuous restraint or intermittent restraint. Furthermore, continuous restraint in step A means that every day during modeling is a restraint day, and the experimental animals are restrained every day using a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period. Or furthermore, intermittent restraint in step A means that every other day during modeling is a restraint day, and the experimental animals are restrained every other day using a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period. Or, the restraint device described in step A is a device that ensures the integrity of the experimental animal's limbs, restricts the movement of the experimental animal but does not affect its breathing. Or, the restraint time described in step A is 3 - 6 hours. Or preferably, the restraint time described in step A is 6 hours. The fasting and water deprivation time in step B is equal to the restraint time. Or for continuous restraint, the modeling time in step C is at least 21 days. Or for intermittent restraint, the modeling time in step C is at least 35 days. Preferably, for intermittent restraint, the modeling time in step C is at least 42 days.

3. The method for constructing a hyperuricemia animal model according to any one of claims 1 or 2, characterized in that, Modeling is performed by restraining experimental animals in combination with a high-purine diet, uricase inhibitor, or uric acid transporter inhibitor.

4. The method for constructing a hyperuricemia animal model according to claim 3, characterized in that, Modeling by restraining experimental animals in combination with a high-purine diet includes the following steps: A. Restraint on restraint days: The experimental animals are restrained by a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period. B. Non-restraint period: The diet is a mixed feed containing dry yeast powder or adenine, and water is freely available. C. The modeling time is at least 21 days.

5. The method for constructing a hyperuricemia animal model according to claim 4, characterized in that, At least one of the following conditions is satisfied: In step B, the content of dry yeast powder in the mixed feed is 5 - 10%. Preferably, in step B, the content of dry yeast powder in the mixed feed is 10%. In step B, the content of adenine in the mixed feed is 0.05 - 0.1%. Preferably, in step B, the content of adenine in the mixed feed is 0.1%. The daily feeding amount is calculated based on the body weight of the experimental animal, with adenine at 125 - 250 mg / kg and dry yeast powder at 12.5 - 25 g / kg. Preferably, the daily feeding amount is calculated based on the body weight of the experimental animal, with adenine at 250 mg / kg and dry yeast powder at 25 g / kg.

6. The method for constructing a hyperuricemia animal model according to claim 3, wherein, Modeling by restraining experimental animals in combination with a uricase inhibitor includes the following steps: A. Restraint on restraint days: The experimental animals are restrained by a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period. B. Non-restraint period: Normal diet, free access to water. C. The modeling time is at least 21 days, and potassium oxonate is intraperitoneally injected in the last 1 - 2 weeks of modeling. Furthermore preferably, at least one of the following conditions is satisfied: In step C, for intraperitoneal injection of potassium oxonate, the daily injection amount is calculated based on the body weight of the experimental animal, with potassium oxonate at 150 - 300 mg / kg. Preferably, in step C, for intraperitoneal injection of potassium oxonate, the daily injection amount is calculated based on the body weight of the experimental animal, with potassium oxonate at 300 mg / kg.

7. The method for constructing a hyperuricemia animal model according to claim 3, wherein Modeling by restraining experimental animals in combination with a uric acid transporter inhibitor includes the following steps: A. Restraint during the restraint period: The experimental animals are restrained with a restraint device for 3 - 6 hours, and water and food are prohibited during the restraint period. B. Non - restraint period: Normal diet, free access to water. C. The modeling time is at least 21 days, and pyrazinamide is administered by gavage in the last 1 - 2 weeks of modeling.

8. The method for constructing a hyperuricemia animal model according to claim 7, wherein At least meet any one of the following: In step C, pyrazinamide is administered by gavage once a day, and the gavage volume is calculated according to the body weight of the experimental animal, with pyrazinamide at 200 - 400 mg / kg. Preferably, pyrazinamide is administered by gavage once a day, and the gavage volume is calculated according to the body weight of the experimental animal, with pyrazinamide at 400 mg / kg.

9. The method for constructing a hyperuricemia animal model according to any one of claims 1-8, characterized in that: The animals used in the construction of the animal model include mammals and avian animals. Further preferably, mammals include primates, rodents, artiodactyls, canids, and felids. Still further preferably, the primates are selected from monkeys and orangutans; the rodents are selected from mice, rats, rabbits, and guinea pigs; the artiodactyls are selected from donkeys, sheep, cows, horses, and pigs; the canids are selected from dogs, wolves, jackals, and foxes; the felids are selected from cats. Further preferably, the avian animals are chickens, ducks, and geese. Preferably, rats or mice are used as experimental animals. Most preferably, mice are used as experimental animals.

10. A hyperuricemia animal model constructed by the construction method described in any one of claims 1 - 9.

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