Construction method of hyperuricemia animal model

Abstract

The invention provides a construction method of a hyperuricemia animal model, which comprises the following steps: in a modeling stage, feeding oteracil potassium to an animal, feeding 10-30% of yeast extract feed to simulate high-purine diet, and exposing in a high-temperature and high-humidity environment to obtain the hyperuricemia animal model. According to the method, a high-temperature and high-humidity environment is constructed, the yeast extract feed is fed to simulate high-purine diet, the daily living environment of coastal residents in the south is replied, and on the basis, oteracil potassium administration is combined to eliminate the influence of uricase, so that the method is close to a human uric acid metabolism mode. The animal model obtained by the invention is relatively higher in blood uric acid level, better in stability and small in renal function damage, the animal always keeps a good mental state in the construction process, and the survival rate is 100% after 12 weeks of modeling. The animal model provided by the invention can provide a scientific basis for subsequent research on hyperuricemia pathogenesis and formulation of prevention and treatment measures under special climate and diet environments, and has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of animal model construction, and in particular to a method for constructing a hyperuricemia animal model. Background Art

[0002] Hyperuricemia is a metabolic disease caused by disorders of purine metabolism in the body. Currently, the overall prevalence rate shows an increasing trend year by year, and the onset is getting younger. Moreover, hyperuricemia is an independent risk factor for various cardiovascular risk events and related diseases. However, its pathogenesis has not been fully elucidated. Establishing a reliable hyperuricemia animal model is the basis for studying its pathogenesis, related complications and therapeutic interventions. At present, the methods for establishing hyperuricemia animal models are complex and diverse, but there is no unified standard. Currently, inducing hyperuricemia with various chemical drugs is the mainstream method for constructing hyperuricemia animal models. According to their different effects on uric acid synthesis metabolism and excretion pathways, they are mainly divided into increasing uric acid sources (adenine, hypoxanthine, fructose, yeast extract), inhibiting uric acid excretion (ethambutol, pyrazinamide), and inhibiting uric acid metabolism method (potassium oxonate). Currently, hyperuricemia shows an aggregated high-incidence status in the southern coastal areas. Therefore, the impact of environmental factors on the body's uric acid metabolism cannot be ignored. The pathogenesis of human hyperuricemia involves multiple factors. Therefore, the combined modeling method is usually used to narrow this gap, but the complex pathogenesis of human hyperuricemia still cannot be fully simulated. When simulating and studying the establishment of a hyperuricemia animal model in the special environment of the southern coast, it is particularly necessary to consider the reproduction of the living environment and dietary conditions.

[0003] The animals selected for hyperuricemia models mainly include rodents, birds and primates. Among rodents, considering the large fluctuations in uric acid levels in female mice and the poor body functions of old mice, adult male mice are generally selected as the modeling objects. However, different from humans, rodents have a uricase gene in their bodies, which causes uric acid to be further decomposed into allantoin and excreted out of the body. Therefore, there are significant differences in blood uric acid levels between rodents and humans. The avian animals mainly selected are chickens and quails. Birds also lack the uricase gene and are ideal hyperuricemia animal models. However, due to species problems and relatively difficult breeding management, their clinical application and transformation are limited. Other animals such as primates are the optimal modeling animals, but they are costly. In addition, there are reports of successfully establishing hyperuricemia or gout models in animals such as tree shrews, zebrafish, and silkworms. However, due to species problems and breeding condition limitations, they are not used as conventional hyperuricemia model animals. Despite the above-mentioned defects, due to the advantages of easy management of rodents, and being homologous to humans in species and relatively close in physiological and biochemical characteristics, they are currently the most commonly used modeling animals.

[0004] Therefore, there is a need for a type of mouse with good stability and little renal function damage, while simulating the hyperuricemia animal model induced by the high temperature, high humidity and high-purine diet environment in the southern coast.

[0005] In view of this, the present invention is hereby provided. Summary of the Invention

[0006] An object of the present invention is to provide a method for constructing an animal model of hyperuricemia, which can be used to construct an animal model of hyperuricemia induced by simultaneously simulating the high-temperature and high-humidity environment and high-purine diet in the southern coastal areas.

[0007] In a first aspect of the present invention, there is provided a method for constructing an animal model of hyperuricemia, comprising the following steps: administering potassium oxonate to an animal during the modeling stage, feeding the animal with a 10-30% yeast extract feed to simulate a high-purine diet, and exposing the animal to a high-temperature and high-humidity environment to obtain an animal model of hyperuricemia.

[0008] Preferably, the animal is fed with a 20% yeast extract feed to simulate a high-purine diet, and the feed is freely available without measurement. Through experimental verification, the feed containing 20 wt% yeast extract can better simulate a high-purine diet.

[0009] Preferably, the administration method of potassium oxonate is intragastric administration, and the dosage of potassium oxonate is 200-300 mg / kg, more preferably 250 mg / kg.

[0010] Preferably, the potassium oxonate is a suspension, and the suspension is prepared using a 0.4-0.6% sodium carboxymethylcellulose solution as a solvent.

[0011] Preferably, the high-temperature and high-humidity environment is simulated using an incubator.

[0012] Preferably, the temperature of the high-temperature and high-humidity environment is 37±0.5°C, the relative humidity is 80±1%, and the treatment duration is 1 h / d.

[0013] Preferably, during the modeling stage, potassium oxonate is intragastrically administered to the animal every morning, and the animal is exposed to the high-temperature and high-humidity environment in an incubator every afternoon. More preferably, the animal is intragastrically treated with potassium oxonate at a fixed time (08:00-10:00) every morning to avoid large fluctuations in blood uric acid levels, and the animal is placed in the incubator for exposure to the high-temperature and high-humidity environment at a fixed time (14:00-16:00) every afternoon.

[0014] Preferably, the daily feeding conditions during the modeling stage are a temperature of 22-26°C and a humidity of 40-60%.

[0015] Preferably, the modeling time is 2-12 weeks.

[0016] Preferably, the animals are adaptively raised for 5 to 9 days before starting the modeling. The raising conditions are a temperature of 22 to 26 °C, a humidity of 40 to 60%, and a day-night alternating lighting time of 11 to 13 h.

[0017] Preferably, the animal includes any one of rats or mice; more preferably, the animal is a rat, and the rats include but are not limited to SD rats, Wistar rats, Lewis rats, etc.

[0018] In a specific embodiment, the rat is an adult male rat, and the age of the adult rat is 6 to 7 weeks.

[0019] In a second aspect of the present invention, there is provided an animal model obtained by using the above construction method.

[0020] In a third aspect of the present invention, there is provided the application of the animal model obtained by the above construction method in studying the molecular mechanism of the occurrence and development of hyperuricemia in residents living in a high-temperature and high-humidity environment with a high-purine diet in the southern coastal areas, or in preparing and / or screening drugs for preventing and / or treating hyperuricemia, or in optimizing treatment regimens.

[0021] The present invention has at least the following beneficial effects:

[0022] (1) The present invention provides a construction method for a hyperuricemia rat model induced by a high-temperature and high-humidity environment, a high-purine diet and a chemical drug. Specifically, a hyperuricemia rat model is established by simulating a high-temperature and high-humidity environment treatment combined with a high-purine diet and intragastric administration of the chemical drug potassium oxonate. The method of the present invention successfully established a hyperuricemia rat model with a significantly higher blood uric acid level than that of the blank control group rats at 2 weeks of modeling. And continuous modeling found that compared with the traditional modeling method of combining potassium oxonate with a high-purine diet, the blood uric acid level of the rats in this composite factor (high-temperature and high-humidity environment exposure + potassium oxonate + high-purine diet) model is relatively higher and the stability is better.

[0023] (2) The hyperuricemia rat model established by the modeling method of the present invention can be further applied to the research on the pathophysiological mechanism, molecular biology and drug treatment of hyperuricemia caused by high-temperature and high-humidity environment and high-purine diet factors in residents in the southern coastal areas of our country.

[0024] (3) The improvement of the present invention lies in innovatively proposing an environmental feature and dietary factors that have not been taken seriously in previous studies to establish a unique hyperuricemia rat model. The rats are treated by highly simulating a high-temperature and high-humidity environment and a high-purine diet, and a maintenance dose of the traditional chemical drug potassium oxonate is used to exert its effect of inhibiting uricase. Different from the models induced by chemical drugs combined with high-purine diet in the past, the hyperuricemia rat model established by the modeling method of the present invention has a more stable blood uric acid level, and has always maintained a good mental state during the treatment process of the high-temperature and high-humidity environment, without heat stroke symptoms such as coma, shock, and respiratory failure. The survival rate after 12 weeks of modeling is 100%.

[0025] (4) The present invention constructs a relatively stable hyperuricemia rat model without severe kidney injury through a high-temperature and high-humidity environment, a high-purine diet combined with the uricase inhibitor potassium oxonate. This model can provide a certain scientific basis for subsequent research on the pathogenesis of hyperuricemia under special high-temperature and high-humidity, high-purine diet environments and the formulation of preventive diagnosis and treatment measures; the method of modeling is convenient, the breeding cost is low, the market demand is large, and it has good application prospects. Brief Description of the Drawings

[0026] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0027] Figure 1 This shows the histopathological changes of the kidney tissue of the hyperuricemia rat model provided by the present invention (100μm) (HE staining); among them, the black arrow: dilation of renal tubules; the red arrow: swelling of renal tubular epithelial cells and stenosis of the lumen; CON: blank control group; HPD: potassium oxonate + high-purine diet group; HTH: high-temperature and high-humidity + potassium oxonate group; CF: high-temperature and high-humidity + potassium oxonate + high-purine diet group.

[0028] Figure 2 This shows the histopathological changes of the kidney tissue of the hyperuricemia rat model provided by the present invention (50μm) (HE staining); among them, the black arrow: dilation of renal tubules; the red arrow: swelling of renal tubular epithelial cells and stenosis of the lumen; CON: blank control group; HPD: potassium oxonate + high-purine diet group; HTH: high-temperature and high-humidity + potassium oxonate group; CF: high-temperature and high-humidity + potassium oxonate + high-purine diet group. Detailed Embodiments

[0029] It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this application belongs.

[0030] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly indicates otherwise, the singular form also includes the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, they indicate the presence of features, steps, operations, devices, components, and / or combinations thereof.

[0031] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0032] Based on the existing key laboratories of the whole army and cooperative laboratories in our hospital, the present invention relies on the following various platforms and scientific research equipment:

[0033] Medical Experiment Section: It has 12 professional laboratories including molecular biology, cell biology, experimental pathology, ultrastructure, standardized PCR, biosensing, immunology, genetics, etc. It has an experimental site of 2,500 square meters and more than 200 experimental instruments with a total value of more than 8 million yuan. It has systematically established main technical platforms in the research fields of histopathology, cell biology, molecular biology, protein chemistry, immunology, biosensing, etc. It can carry out experimental techniques such as fluorescence quantitative PCR, two-dimensional protein electrophoresis, electron microscopy, flow cytometry, stem cell culture, ultrastructural pathology, etc.

[0034] Animal Experiment Center: The existing experimental facilities can undertake experiments on ordinary-grade minipigs, dogs, guinea pigs, rabbits, rats, mice, guinea pigs, rabbits in barrier environments. Guangdong Experimental Animal Use License Number: SYXK(Guangdong)2014 - 0100, Military Barrier Environment Experimental Animal Use License Number: YXK(Military)2012 - 0056, Military Ordinary Environment Experimental Animal Use License Number: SYXK(Military)2012 - 0055. Micro CT, non-invasive blood pressure measuring instrument for rats and mice, high-frequency electrotome, automatic biochemical analyzer, small animal ventilator, microsurgical operating microscope, high-temperature and high-humidity incubator, centrifuge, biological safety cabinet, isolator, IVC, high-pressure sterilizer, all-round monitor, nine-hole shadowless operating lamp, stainless steel clean operating table.

[0035] Embodiment

[0036] This embodiment provides a method for constructing a hyperuricemia rat model by simulating the high-temperature and high-humidity environment, high-purine diet and chemical drugs in the southern coastal areas, including the following steps:

[0037] Step (1): Preparation of animals: 24 clean-grade male Sprague-Dawley rats (hereinafter referred to as SD rats), 6-7 weeks old, with a body weight of 150-200 g.

[0038] Step (2): Establishment of the hyperuricemia rat model: 24 SPF-grade SD rats were randomly divided into a blank control group, a high-temperature and high-humidity + potassium oxonate group, a potassium oxonate + high-purine diet group, and a high-temperature and high-humidity + potassium oxonate + high-purine diet group, with 6 rats in each group. The animals in each group were housed separately, and the lighting in the breeding room was cycled alternately for 12 / 12 h day and night. The environmental temperature was maintained at 22-26 °C and the humidity was 40-60%. After 1 week of adaptive feeding of SD rats, the modeling experiment was officially carried out.

[0039] The rats in the high-temperature and high-humidity + potassium oxonate + high-purine diet group were intragastrically administered with 250 mg / kg potassium oxonate suspension (the suspension was prepared with 0.5% sodium carboxymethylcellulose solution as the solvent) at a fixed time (09:00) every morning, and were transferred to a specific constant-temperature incubator for treatment at a fixed time (15:00) in the afternoon. The temperature in the incubator was set at 37 ± 0.5 °C and the relative humidity was 80 ± 1%, and the treatment duration was 1 h / d. The conditions of the culture environment at other times were: temperature 22-26 °C, humidity 40-60%. They were fed with 20% yeast extract feed without time limit and without measurement.

[0040] The rats in the high-temperature and high-humidity + potassium oxonate group were intragastrically administered with 250 mg / kg potassium oxonate suspension (the suspension was prepared with 0.5% sodium carboxymethylcellulose solution as the solvent) at a fixed time (09:00) every morning, and were transferred to a specific constant-temperature incubator for treatment at a fixed time (15:00) in the afternoon. The temperature in the incubator was set at 37 ± 0.5 °C and the relative humidity was 80 ± 1%, and the treatment duration was 1 h / d. The conditions of the culture environment at other times were: temperature 22-26 °C, humidity 40-60%.

[0041] The rats in the potassium oxonate + high-purine diet group were intragastrically administered with 250 mg / kg potassium oxonate suspension every morning. They were fed with 20% yeast extract feed without time limit and without measurement.

[0042] The blank control group was intragastrically administered with the same dose of 0.5% sodium carboxymethylcellulose solution every morning.

[0043] Step (3): Rectal temperature measurement time and method: During the period when the rats in the high temperature and high humidity + oxyonic acid potassium group were in the climate chamber, all 4 groups of rats were uniformly fasted and water-deprived. Rectal temperature was measured once at the 0th min, 20th min, 40th min, and 60th min, with the measurement depth of 3 cm and the measurement duration of at least 1 min. Ensure sufficient oxygen in the incubator and continuously observe the mental state and vital signs of the rats.

[0044] Step (4): Measurement and calculation method of dehydration rate: Before the treatment of the rats in the high temperature and high humidity + oxyonic acid potassium group and the high temperature and high humidity + oxyonic acid potassium + high purine diet group, the body weights of all rats were measured; after the end, the body weights of all rats were measured again. Among them, for the two groups of rats exposed to high temperature and high humidity, the body surface moisture was wiped clean with absorbent gauze before weighing. Dehydration rate = (body weight before treatment - body weight after treatment) / body weight before treatment.

[0045] Step (5): Collection of blood samples: At the 0th, 2nd, 4th, 6th, 8th, 10th, and 12th weeks of the experiment, all rats were fasted for 12 h without water deprivation, and 0.5 ml of blood was collected from the tail to detect the serum uric acid level to evaluate the success of the model; the serum creatinine level was detected to evaluate the degree of renal damage of the model.

[0046] Serum uric acid detection method: After the blood specimen was left standing at room temperature for 30 min, it was centrifuged at 3500 rpm at 4 °C for 15 min to take the upper serum. An uric acid assay kit was used and operated according to the instructions and the guidance of the manufacturer to determine the distribution of the samples on the 96-well plate. Sample addition: ① Blank well: Add 10 μL of distilled water to the blank control well; ② Standard well: Add 10 μL of standard solution (concentration: 400 μmol / L); ③ Sample well: Add 10 μL of the serum sample of the rat to be tested; then add 250 μL of reaction solution (prepared from Tris-HCl buffer, peroxidase, and uricase) to each well. Cover the sealing film and gently mix, and then incubate in an incubator at 37 °C for 10 min. The absorbance value OD of each well was measured sequentially by an enzyme-labeled instrument under the condition of an ultraviolet wavelength of 510 nm. According to the concentration of the standard product and the absorbance values of the blank well, standard well, and sample well, the corresponding serum uric acid value of the sample was calculated. The formula is as follows:

[0047] Sample uric acid concentration (μmol / L) = standard product concentration 400 μmol / L * (OD sample - OD blank) / (OD standard - OD blank);

[0048] Serum creatinine detection method: Use a creatinine assay kit to evaluate renal function impairment. Specifically, operate according to the instructions and the guidance of the manufacturer to determine the distribution of samples on the 96-well plate; Sample addition: ① Blank wells: Add 6 μL of distilled water to the blank control wells; ② Standard wells: Add 6 μL of standard solution (concentration 442 μmol / L); ③ Sample wells: Add 6 μL of the serum sample of the rat to be tested. Then add 180 μL of enzyme solution A to each well and incubate in a 37 °C incubator for 5 min. Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance value A1 at a wavelength of 546 nm; Add 60 μL of enzyme solution B to each well and sequentially measure the absorbance value A2 of each well under the condition of 546 nm wavelength of the ELISA reader. Calculate the corresponding serum creatinine value of the sample according to the concentration of the standard product and the absorbance values of the blank wells, standard wells, and sample wells. The formula is as follows:

[0049] Sample creatinine concentration (μmol / L) = Standard product concentration 442 μmol / L * (A2 sample - A1 sample * K) - (A2 blank * - A1 blank * K) / (A2 standard product - A1 standard product * K) - (A2 blank * - A1 blank * K);

[0050] [Note: Dilution factor K = (Sample addition volume + Volume of enzyme solution A) / (Sample addition volume + Volume of enzyme solution A + Volume of enzyme solution B) = 186 / 246].

[0051] Step (6) Observation of renal pathological changes: Animal sampling and sample preparation: At 12 weeks of modeling in this experiment, anesthetize each group of rats with 30 mg / kg of 3% sodium pentobarbital, dissect and take the ipsilateral kidney, cut it evenly into 3 pieces, and immerse them in 4% paraformaldehyde fixative within 1 min. Embed the kidney specimens fixed with 4% paraformaldehyde in paraffin, then perform serial sectioning, and perform hematoxylin-eosin (HE) staining to observe renal pathological changes.

[0052] HE staining technique: After the tissue is fixed with 4% paraformaldehyde, embed it in paraffin, trim the wax block, continuously cut 3-μm tissue sections with a microtome. After placing them in a 60 °C oven for 1 h for baking, immediately immerse them in xylene for dewaxing, 20 min × 2 times. After hydrating with 90%, 80%, and 70% gradient alcohol for 5 min each, immerse them in the modified hematoxylin staining solution for 6 min, differentiate with 1% hydrochloric acid alcohol for 5 s, rinse with running water to blue for 10 min, then immerse them in eosin staining solution for 3 min, wash with 95% alcohol for 1.5 min × 2 times, dehydrate with gradient alcohol, soak in xylene for 5 min × 3 times for clearing, and seal with neutral gum and dry, then observe and take pictures under a BX-51 microscope.

[0053] Step (7) Statistical analysis method: IBM SPSS 25.0 statistical software was used for data analysis. Measurement data were expressed as [description of measurement data representation]. If they conformed to the normal distribution test and had homogeneous variance, parametric test methods were used. One-way ANOVA was used to compare the serum uric acid and creatinine values between different groups. If the one-way ANOVA showed statistically significant differences, the LSD-test was used for multiple comparisons. P < 0.05 indicated statistically significant differences.

[0054] The results of the serum uric acid values of the rat models are shown in Table 1. It can be seen from the results that compared with the blank control group, the potassium oxonate + high-purine diet group, and the high temperature and high humidity + potassium oxonate group, the hyperuricemia rat model established by the modeling method of the high temperature and high humidity + potassium oxonate + high-purine diet group had a more stable serum uric acid level. At the 2nd week of the experiment, the serum uric acid level reached the highest value, and then showed a gradually decreasing trend. However, the serum uric acid level was still higher than that of the other three groups of rats at the 12th week.

[0055] Table 1 Changes in serum uric acid values of rats in each group at different time points of the experiment

[0056]

[0057] Note: (1) Compared with the CON group, aP < 0.05; compared with the HPD group, bP < 0.05. (2) At 8 weeks, the results of the normal distribution test of the uric acid values in different groups were W = 0.878, P = 0.044 < 0.05, not conforming to the normal distribution. The results of the homogeneity of variance test were L = 0.402, P = 0.803 > 0.05, meeting the requirements of homogeneous variance. The data were expressed as M(P25 - P75). The non-parametric test Kruskal-Wallis H method was used for comparison among multiple groups, H = 12.100, P = 0.017 < 0.05, with statistically significant differences. The Bonferroni correction test method was used for multiple comparisons. The comparison between the CF group and the CON group showed P = 0.035 < 0.05, with statistically significant differences. (3) At 0 weeks, 2 weeks, 4 weeks, 6 weeks, 10 weeks, and 12 weeks, the serum uric acid data of the four groups conformed to the normal distribution test and had homogeneous variance. (4) According to the above results, the one-way ANOVA test method was used to compare the serum uric acid values of the four groups of rats at different times. The F value and P value are shown in the above table. The LSD-test was used for multiple comparisons. P < 0.05 indicated statistically significant differences. (5) Number of cases: At 0 weeks, 2 weeks, 4 weeks, and 6 weeks of the experiment, there were 6 rats in each group. At 8 weeks, 10 weeks, and 12 weeks of the experiment, there were 3 rats in each group. CON: Blank control group; HPD: Potassium oxonate + high-purine diet group; HTH: High temperature and high humidity + potassium oxonate group; CF: High temperature and high humidity + potassium oxonate + high-purine factor group.

[0058] The serum creatinine values of the rat models are shown in Table 2. The changing trend of the serum creatinine levels of the rats was observed for 12 weeks. The serum creatinine levels of all the rats in each group showed an overall upward trend. In the high temperature and high humidity + potassium oxonate + high purine diet group, the serum creatinine levels were higher than those of the other three groups at the 2nd and 6th weeks of the experiment. At the 10th and 12th weeks of the experiment, there were no significant statistical differences in the serum creatinine levels compared with those of the rats in other groups, indicating that no obvious acute renal failure occurred during the experiment.

[0059] Table 2 Changes in serum creatinine values of rats in each group at different time points of the experiment

[0060]

[0061] Note: (1) Compared with the CON group, aP < 0.05; compared with the HPD group, bP < 0.05. (2) The results of the normality test of the serum creatinine values at 0 week were W = 0.848, P = 0.016 < 0.05, not following a normal distribution. The results of the homogeneity of variance test were L = 0.845, P = 0.510 > 0.05, meeting the requirements of the homogeneity of variance test. The data were expressed as M(P25 - 75). The non-parametric test Kruskal-Wallis H method was used for comparison among multiple groups, K = 4.516, P = 0.341 > 0.05, and the differences were not statistically significant. (3) The serum creatinine data at 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks, and 12 weeks conformed to the normality test and the homogeneity of variance. (4) According to the above results, the one-way ANOVA test method was used to compare the serum creatinine values at different times, and the F value and P value are shown in the above table. (5) Number of cases: There were 6 rats in each group at 0 week, 2 weeks, 4 weeks, and 6 weeks of the experiment, and 3 rats in each group at 8 weeks, 10 weeks, and 12 weeks of the experiment. CON: blank control group; HPD: potassium oxonate + high purine diet group; HTH: high temperature and high humidity + potassium oxonate group; CF: high temperature and high humidity + potassium oxonate + high purine factor group.

[0062] At 12 weeks of the experiment, each group of rats was deeply anesthetized and the kidneys were taken to observe their histopathological changes. The results are shown in Figure 1-2 , and it can be seen from the results that after HE staining, under light microscopy, the renal tissue morphology of the blank control group was basically normal, the glomerular morphology was unchanged, the renal tubules were not dilated, and there was no inflammatory cell infiltration. In the potassium oxonate + high purine diet group, a small amount of glomerular parietal epithelial cells and renal tubular epithelial cells were necrotic and exfoliated, the renal tubular lumen was dilated, and the wall was thinned. In the high temperature and high humidity + potassium oxonate group, in addition to inflammatory infiltration in the cell gap, swelling of the proximal tubular epithelial cells occurred, and some lumens were significantly narrowed. In the high temperature and high humidity + potassium oxonate + high purine diet group, significant inflammatory infiltration in some tubular spaces, swelling of the proximal tubular epithelial cells, and narrowing of the lumen space were observed. In addition, atrophy of some renal tubular epithelial cells and shedding of microvilli resulting in lumen dilation were also present, but no obvious cell necrosis was seen, and the renal function was not severely affected.

[0063] In addition, the rats in the high temperature and high humidity + potassium oxonate + high purine diet group maintained a good mental state during the high temperature and high humidity treatment process, without heat stroke symptoms such as coma, shock, and respiratory failure, as well as severe kidney damage. The survival rate was 100% after 12 weeks of modeling.

[0064] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for constructing an animal model of hyperuricemia, characterized in that, It includes the following steps: During the modeling stage, potassium oxonate is administered to animals, and they are fed a 10-30% yeast extract diet to simulate a high-purine diet and placed in a high-temperature and high-humidity environment for exposure to obtain a hyperuricemia animal model.

2. The method for constructing a hyperuricemia animal model according to claim 1, wherein They are fed a 20% yeast extract diet to simulate a high-purine diet and allowed to eat freely without measurement.

3. The method for constructing a hyperuricemia animal model according to claim 1, characterized in that, The administration method of potassium oxonate is intragastric administration, and the dosage of potassium oxonate is 200-300 mg / kg.

4. The method for constructing a hyperuricemia animal model according to claim 1, characterized in that, The potassium oxonate is a suspension, and the suspension is prepared using a 0.4-0.6% sodium carboxymethylcellulose solution as a solvent.

5. The method for constructing a hyperuricemia animal model according to claim 1, characterized in that, The temperature of the high-temperature and high-humidity environment is 37±0.5°C, the relative humidity is 80±1%, and the treatment duration is 1 h / d.

6. The method for constructing a hyperuricemia animal model according to claim 1, wherein The daily feeding conditions during the modeling stage are a temperature of 22-26°C and a humidity of 40-60%.

7. The method for constructing a hyperuricemia animal model according to claim 1, wherein, The modeling time is 2-12 weeks.

8. The method for constructing a hyperuricemia animal model according to claim 1, characterized in that, The animals are adaptively fed for 5-9 days before starting the modeling, and the feeding conditions are a temperature of 22-26°C, a humidity of 40-60%, and a day-night alternating lighting time of 11-13 h.

9. The method for constructing a hyperuricemia animal model according to claim 1, characterized in that, The animals include any one of rats or mice.

10. The application of the animal model obtained by the construction method according to any one of claims 1-9 in studying the molecular mechanism of the occurrence and development of hyperuricemia in residents living in the high-temperature, high-humidity and high-purine diet living environment in the southern coastal areas, or in preparing and / or screening drugs for preventing and / or treating hyperuricemia, or in optimizing treatment regimens.

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