CN115039737B - Method for establishing uronate kidney deposition animal model - Google Patents
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Feed For Specific Animals (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a method for establishing an animal model for urate renal deposition, which comprises the following steps: rats were used as experimental animals; the kidney deposition of the uronate of the experimental animal is induced by combining the potassium oxazinate, adenine, yeast extract powder and lipopolysaccharide. The invention builds an animal model of urate renal deposition, preliminarily defines the pathological characteristics of the animal model, and can provide experimental foundation for the pathological mechanism of urate renal deposition and prevention and treatment research.
Description
Technical Field
The invention provides a method for establishing an animal model for uric acid salt renal deposition, and belongs to the technical field of animal experiment models.
Background
Urate renal deposition is a uric acid metabolic disorder disease in which blood uric acid in the body is supersaturated and deposited in renal tissues, and secondary kidney pathological changes are caused, and is closely related to hyperuricemia, and can cause secondary tissue injury and induce joint gout. Effective intervention of the urinary acid salt renal deposition has important significance for blocking the progress of hyperuricemia and preventing and treating joint gout. However, screening research of pathological mechanisms of urate renal deposition and prevention and treatment drugs is greatly dependent on establishment and application of an animal model of urate renal deposition.
However, no clear research on an animal model of urate renal deposition is currently known, only relevant scattered descriptions are found in reports of animal models of renal injury, and the modeling method and the tissue positioning and evaluation of urate renal deposition have certain limitations. Such as: (1) because birds lack uricase that breaks down uric acid, their blood uric acid levels are elevated more markedly than in conventional rodents, and binding to hyperuricemia is an important biochemical basis for urate deposition, birds become a dominant animal model of uric acid metabolic disorders including urate renal deposition. However, the kidney tissue structure of the poultry is greatly different from that of human beings, and the pathological discussion of the poultry may have limitation in the prevention and treatment research of urate renal deposition. (2) Kidney tissue is composed of kidney parenchyma and renal pelvis, kidney parenchyma comprises cortex and medulla, and the definition of the positioning characteristics of urate deposition in kidney tissue is the basis of pathology and prevention and treatment research, however, no related research report is seen at present. (3) Urate deposition is easy to dissolve in water, and the current literature reports that the conventional hematoxylin-eosin (HE) staining procedure has a water-soluble step, so that the actual condition of urate deposition is difficult to accurately show. And urate deposition has tissue distribution nonuniformity, and a quantitative evaluation method of the deposition is yet to be explored and defined.
The limitations in animal model research greatly limit the basic research and clinical control of urate renal deposition, so the establishment of a proper animal model is a key problem to be solved urgently in the field.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention aims to provide a method for establishing an animal model for urate renal deposition; the invention also aims to provide an inducer for establishing an animal model of uronate renal deposition.
As one aspect of the present invention, the present invention provides a method of constructing an animal model of uronate renal deposition, the method comprising the steps of:
selecting a rat as a model animal;
and (3) administering an inducer to the experimental animal, wherein the inducer consists of potassium oxazinate, adenine, yeast extract and lipopolysaccharide.
In a specific embodiment, the potassium oxazinate, adenine and yeast extract in the inducer are administrated by lavage, the administration dosages are respectively 1g/kg, 80mg/kg and 10g/kg of each experimental animal, and the lipopolysaccharide is administrated by intraperitoneal injection, and the administration dosages are 0.2 mg/kg-1.2 mg/kg.
In a specific embodiment, each experimental animal in the inducer is administered a lipopolysaccharide dosage of 0.2mg/kg to 1.2mg/kg; preferably, each experimental animal in the inducer is administered a lipopolysaccharide dose of 0.2mg/kg.
As another aspect of the present invention, the present invention provides an inducer for an animal model of renal deposition of uronate, which consists of potassium oxazinate, adenine, yeast extract, lipopolysaccharide.
In a specific embodiment, the potassium oxazinate, adenine and yeast extract in the inducer are administrated by lavage, the administration dosages are respectively 1g/kg, 80mg/kg and 10g/kg of each experimental animal, and the lipopolysaccharide is administrated by intraperitoneal injection, and the administration dosages are 0.2 mg/kg-1.2 mg/kg.
In a specific embodiment, each experimental animal in the inducer is administered a lipopolysaccharide dosage of 0.2mg/kg to 1.2mg/kg; preferably, each experimental animal in the inducer is administered a lipopolysaccharide dose of 0.2mg/kg.
The invention can effectively shape the uronate kidney deposition animal model, and can provide experimental foundation for clinical uronate kidney deposition pathological mechanism and prevention and treatment research.
Drawings
FIG. 1 is a chart of hexamine silver staining of kidney tissue sections of rats in example 1.
FIG. 2 is a bar graph of the blood uric acid levels of the rats in example 3.
FIG. 3 is a bar graph of uric acid clearance for each group of rats in example 3.
FIG. 4 is a bar graph of the serum creatinine for each group of rats in example 3.
Fig. 5 is a bar graph of creatinine clearance for each group of rats in example 3.
FIG. 6 is a bar graph of serum beta 2-microglobulin levels for each group of rats in example 3.
FIG. 7 is a bar graph of serum cystatin C levels for each group of rats in example 3.
FIG. 8 is a hematoxylin-eosin staining chart of kidney tissue sections of rats of each group in example 3.
Detailed Description
The present invention will be further described with reference to the following specific embodiments, which are, however, not limited to the following embodiments.
EXAMPLE 1 screening study of the inducer species of the rat model of uronate renal deposition
1. The purpose of the experiment is as follows: screening of inducers for inducing rat urate renal deposition
2. Experimental method
2.1 grouping of animals
The total number of male SD rats is 40, SPF grade, weight is 180+ -10 g, after 3 days of adaptive feeding, the male SD rats are randomly divided into 4 groups according to body mass, namely a normal group, an inducer group A, an inducer group B and an inducer group C, wherein 10 induction agents in each group are respectively formed, and the induction methods of the induction agents in each group are shown in table 1, and are all administered singly.
Table 1 experiment groups of inducer compositions and induction methods
2.2 obtaining materials
After molding, urine of the experimental animal for 12 hours is collected by a metabolism cage method, and the urine is measured. Abdominal aortic blood was taken after anesthetizing the experimental animals and serum was isolated. The experimental animals were sacrificed, the abdominal cavity was exposed, kidneys were separated, and kidney tissues were dissected longitudinally and fixed in absolute ethanol.
2.3 uric acid salt renal deposition hexamine silver staining and data analysis
Preparing paraffin sections from kidney tissues fixed by absolute ethyl alcohol, and dyeing the kidney tissue sections by adopting a hexamine silver method, wherein urate deposition is black. And acquiring images under an inverted microscope, selecting non-overlapping visual fields, and comprehensively acquiring deposition images of all the sections. Image J software measured the deposition area of each picture and summed up the urate deposition area of each kidney tissue section for statistics.
3. Data analysis
Statistical analysis was performed using GraphPad prism7.0 software, and data were taken as mean ± standard deviation The data comparison between groups is expressed by selecting one-factor analysis of variance or Kruskal-Wallis anecdotal test according to normal and variance of each group, and the comparison between groups is based on variance of each group and is selected by Dunnett-t test or Dunnett's T3 test according to variance of each group, and P is calculated by<A difference of 0.05 is statistically significant.
4. Main instruments and equipment (see Table 1 below)
TABLE 1 Main instruments and apparatus
| Name of the name | Model number | Manufacturer' s |
| Freezing embedding machine | KH-BL | HUBEI XIAOGAN KUOHAI MEDICAL TECHNOLOGY Co.,Ltd. |
| Paraffin tissue slicer | Reichert Histo STAT | AO Co Ltd |
| Microscope | Olmpus BX53 | Orinbas Corp Japan |
| Camera with camera body | DP72CCD | Orinbas of Japan |
5. Main reagents and drugs (see Table 2 below)
TABLE 2 Main reagents and drugs
6. Results
6.1 uric acid levels of renal deposition
The staining results of the kidney tissue sections show that uric acid salt deposition is not found in the kidney tissue sections of rats in the normal group and the rat kidney tissue sections in the B induction group, a small amount of black uric acid salt deposition is visible in the kidney tissue sections of 2/10 rats in the A induction group, and obvious uric acid salt deposition is visible in the kidney tissue sections of rats in the C induction group. The specific results are shown in Table 3.
Table 3 rat uronate renal deposition levels in groups
6.2 pathological observations of urate renal deposition
The silver staining pattern of the kidney tissue hexamine is shown in fig. 1. The results showed that no urate deposition was seen in the cortical, medulla and papillary portions of kidney tissue of both normal and B-induced rats. A small amount of urate deposition is visible in the papillary portion of kidney tissue of the group A induced rats, and no urate deposition is visible in the renal cortex and renal medulla portions. Obvious urate deposition can be seen in the cortex, medulla and papilla parts of kidney tissues of rats in the induction group C, and the urate deposition area is that the renal papilla part is greater than the renal medulla and greater than the renal cortex.
7. Analysis of results
The present study screened inducers of animal models of urate renal deposition. Urinary acid salt renal deposition is a pathological state in which blood uric acid in the body is supersaturated and deposited in renal tissues, and hyperuricemia is the biochemical basis. Meanwhile, kidney is an important organ for uric acid excretion, and pathological changes of kidney are also important factors for uric acid renal deposition. Thus, the present study developed an animal model of uronate renal deposition from both the aspects of promoting elevated blood uric acid and inducing renal injury.
The chemical inducer used for increasing blood uric acid at present mainly comprises potassium oxyzinate, ethambutol and adenine, and in addition, high yeast diet and high fructose diet can also cause the increase of blood uric acid level. Wherein the potassium oxazinate is uricase inhibitor, and only aims at animals, such as rodents, large and mice, in which uricase exists, and uric acid excretion is influenced by inhibiting uric acid from decomposing into soluble substances. Ethambutol is an antituberculosis drug which can raise blood uric acid by inhibiting renal uric acid excretion, but has serious kidney damage of model animals and certain application limitation. Adenine is one of uric acid precursor substances, and promotion of uric acid synthesis is an important mechanism for increasing blood uric acid. Both high yeast diet and high fructose diet are diet inducers that mimic the occurrence of clinical hyperuricemia. Because the mechanism of human blood uric acid rise is complex, the modeling of the model is often multi-method combined modeling at present. Therefore, the study combines early modeling experience, and the yeast extract, adenine and potassium oxazinate are adopted to combine with gastric lavage to induce the rise of blood uric acid level from the aspects of promoting uric acid generation, inhibiting uric acid excretion and inhibiting uricase activity. Lipopolysaccharide is the main component of endotoxin of gram-negative bacteria, is the main inducer for the current research of kidney injury, and the research reports that lipopolysaccharide is used for intraperitoneal injection to induce experimental rat kidney injury according to the literature.
The research result shows that the kidney tissue section of the rat with the B inducer group using lipopolysaccharide intraperitoneal injection to induce kidney injury as a modeling mode does not see urate deposition; only 2/10 kidney tissue sections of rats have small amount of urate deposition in the A inducer group taking yeast extract, adenine and potassium oxazinate combined gastric lavage induced hyperuricemia as a model forming mode; whereas 10/10 of the rat kidney tissue sections showed significant urate deposition in the C inducer group in combination with yeast extract, adenine and potassium oxazinate lavage.
Taken together, the study suggests that 1g/kg of potassium oxazinate, 80mg/kg of adenine, 10g/kg of yeast extract are infused into the stomach, and combined with intraperitoneal injection of 0.2mg/kg of lipopolysaccharide, the rat model of catheter renal deposition can be remarkably induced.
EXAMPLE 2 dose screening study of rat urate renal deposition model inducer
1. The purpose of the experiment is as follows: the influence of the compound inducer containing different doses of lipopolysaccharide on experimental rats is discussed, and the composition of the rat urate renal deposition inducer is further clarified.
2. Experimental method
Male SD rats were classified into 40 groups, each group comprising 10 normal group, C inducer group, D inducer group and E inducer group, each group comprising 180+ -10 g SPF, after 3 days of adaptive breeding, the total body mass was randomly divided into 4 groups, and the inducer composition and induction method of each group were shown in Table 4 and were administered in a single dose. After molding, the general state of each group of experimental rats was observed.
Table 4 experiment groups of inducer compositions and induction methods
3. Main reagents and drugs (see Table 5 below)
TABLE 5 Main Agents and drugs
| Name of the name | Product lot number | Manufacturer' s |
| Oxazinate potassium salt | J30GS139145 | Source leaf biology Co Ltd |
| Adenine (A) | WXBD1877V | Sigma-Aldrich trade Co., ltd |
| Yeast extract powder | 4304391-02 | ThermoFisher Scientific Co Ltd |
| Sodium hydroxymethyl cellulose | Y24S11X125635 | Source leaf biology Co Ltd |
| Lipopolysaccharide | 0000081275 | Beijing Bayer Di Biotechnology Co Ltd |
6. Results
The results show that: after the E induction group rats die, 2/10 rats die, and the other rats have sallow hair, obvious abdomen contraction, arch backs, pile, squint eyes and listless mental state. D, death of the induced rats is not seen after molding, but sallow hair, obvious abdominal contraction, bow back, pile, squinting eyes and listlessness are seen, and symptom relief is not seen after molding for 12 hours. Rats in the induction group C have no death, and after molding for 1h, the abdomen is seen to be contracted and the back is seen to be arched, but after molding for 4h, the state of mind is obviously relieved, and the state of mind is obviously better than that of the D, E inducer group.
7. Analysis of results
Screening studies of rat uronate kidney deposit model inducers in example 1 suggest that hyperuricemia in combination with kidney injury is a two-way important factor in urate kidney deposit. The study selects lipopolysaccharide with concentration of 1.2mg/kg, 0.6mg/kg and 0.2mg/kg respectively for dosage screening of rat model inducer of uronate renal deposition. As a result, it was found that administration of 1.2mg/kg of the model inducer resulted in death of 20% of experimental animals, administration of 0.6mg/kg of the model inducer resulted in severe mental state of experimental animals, and the 12 hours still remained unreliable, and none of the above model inducers was suitable in consideration of the influence of the general state of model animals on the drug study. Whereas administration of 0.2mg/kg of lipopolysaccharide model inducer resulted in significant recovery of the abnormality in the general state of the experimental animals 4 hours after molding. In combination with the status of the experimental animals in this study, and the significant deposition of urate in kidney tissue sections resulting from the administration of 0.2mg/kg lipopolysaccharide inducer as described in example 1, it was suggested that the 0.2mg/kg dose of lipopolysaccharide was the appropriate concentration for modeling the rat uric acid kidney deposition model.
EXAMPLE 3 study of Complex inducer modeling rat urate renal deposition model
1. The purpose of the experiment is as follows: the pathological characteristics of the rat urate renal deposition model are clear.
2. Experimental method
2.1 grouping of animals
Male SD rats were divided into 20 SPF-grade animals, weighing 180+ -10 g, and after 3 days of adaptive feeding, they were randomly divided into normal and model groups according to body mass, each group being 10 animals. The normal group is administrated with CMC-Na solution of 0.5% for stomach irrigation and physiological saline for intraperitoneal injection, and the model group is molded by single intraperitoneal injection of 1g/kg of potassium oxazinate, 80mg/kg of adenine, 10g/kg of yeast extract for stomach irrigation and 0.2mg/kg of lipopolysaccharide.
2.2 obtaining materials
After molding, urine of the experimental animal for 12 hours is collected by a metabolism cage method, and the urine is measured. Abdominal aortic blood was taken after anesthetizing the experimental animals and serum was isolated. Experimental animals were sacrificed, abdominal cavities were exposed, kidneys were isolated, and kidney tissues were dissected longitudinally and fixed in absolute ethanol, 10% formalin, respectively.
2.3 uric acid salt renal deposition hexamine silver staining and data analysis
Preparing paraffin sections from kidney tissues fixed by absolute ethyl alcohol, and dyeing the kidney tissue sections by adopting a hexamine silver method, wherein urate deposition is black. And acquiring images under an inverted microscope, selecting non-overlapping visual fields, and comprehensively acquiring deposition images of all the sections. Image J software measured the deposition area of each picture and summed up the urate deposition area of each kidney tissue section for statistics.
2.4 Biochemical index detection
Uric acid level in serum and urine is detected by uricase method, and uric acid clearance is calculated. Creatinine oxidase method detects creatinine level in serum and urine, and calculates creatinine clearance rate. The beta 2-microglobulin level in serum is detected by a latex immunonephelometry method. Cystatin C levels in serum were detected by latex immunonephelometry.
Uric acid clearance (mL/min) =uric acid level/blood uric acid level× (urine volume/urine collection duration)
Creatinine clearance (mL/min) =urinary creatinine level/blood creatinine level× (urine volume/urine collection duration)
2.5 histopathological observations of the kidneys
Paraffin sections were prepared from 10% formalin-fixed kidney tissue and stained with hematoxylin-eosin stain. An inverted microscope was used to collect images for histopathological observation.
3. Data analysis
Statistical analysis was performed using GraphPad prism7.0 software, and data were taken as mean ± standard deviation The data comparison between the two groups is represented by selecting t-test or t' -test according to the normal state and variance of each group to obtain P<0.05 statistical significance of the differenceMeaning.
4. Main instruments and equipment (see Table 6 below)
TABLE 6 Main instruments and apparatus
| Name of the name | Model number | Manufacturer' s |
| Desk type centrifugal machine | DT5-3 type | Beijing era North centrifuge Co., ltd |
| Freezing embedding machine | KH-BL | HUBEI XIAOGAN KUOHAI MEDICAL TECHNOLOGY Co.,Ltd. |
| Paraffin tissue slicer | Reichert Histo STAT | AO Co Ltd |
| Microscope | Olmpus BX53 | Orinbas Corp Japan |
| Camera with camera body | DP72CCD | Orinbas of Japan |
| Enzyme label instrument | sunrise | TeCAN company Switzerland |
| Water bath kettle | HH-1 high-end type | Jintan city and west sense, laboratory instrumentation factory |
5. Main reagents and drugs (see Table 7 below)
TABLE 7 Main reagents and drugs
6. Results
6.1 uric acid levels of renal deposition
The staining results of kidney tissue sections showed: normal groups of rat kidney tissue sections had no urate deposition. The kidney medulla part of 10/10 rat kidney tissue sections can be seen to have obvious urate deposition, the kidney cortex part can be seen to have a small amount of urate deposition, and the area of urate renal deposition in the statistical model group is (6.74+/-2.19) multiplied by 10 -6 。
6.2 blood uric acid levels and uric acid clearance
Compared with the normal group, the blood uric acid level of rats in the model group is obviously increased (P < 0.01), and the uric acid clearance rate is obviously reduced (P < 0.05). The specific results are shown in Table 8, FIGS. 2-3.
Table 8 blood uric acid levels and uric acid clearance (n=10) of rats in each group
| Group of | Blood uric acid (mu mol/L) | Uric acid clearance (. Times.10) -2 ，mL/min) |
| Normal group | 172.55±67.04 | 9.62±4.91 |
| Model group | 944.76±242.26 ** | 5.55±3.17 * |
Note that: in contrast to the normal group, * P＜0.05。
6.3 blood creatinine levels and creatinine clearance
Creatinine clearance is a common indicator for clinical detection of renal function. Compared with the normal group, the blood creatinine level of the rats in the model group is obviously increased (P < 0.01), and the creatinine clearance rate is obviously reduced (P < 0.01). The specific results are shown in Table 9, FIGS. 4-5.
Table 9 blood creatinine levels and creatinine clearance (n=10) for each group of rats
| Group of | Blood creatinine (mu mol/L) | Creatinine clearance (mL/min) |
| Normal group | 17.12±4.56 | 0.71±0.26 |
| Model group | ** | ** |
Note that: in contrast to the normal group, ** P＜0.01。
6.4β2-microglobulin and cystatin C levels
Beta 2-microglobulin and cystatin C are sensitive indexes for evaluating the functions of renal tubules and glomeruli respectively. Compared with the normal group, the serum beta 2-microglobulin level of the rats in the model group is obviously increased (P is less than 0.01), and the serum cystatin C level of the rats in the model group is not statistically different. The specific results are shown in Table 10, FIGS. 6-7.
Table 10 levels of rat β2-microglobulin and cystatin C for each group (n=10)
| Group of | Beta 2-microglobulin (mg/mL) | Cystatin C (mg/mL) |
| Normal group | 180.50±83.18 | 598.66±270.37 |
| Model group | ** | 607.50±251.16 |
Note that: in contrast to the normal group, ** P＜0.01。
6.5 pathological observations of kidney tissue
Hematoxylin-eosin staining of kidney tissue sections of each group of rats is shown in fig. 8. The results show that: the kidney tissue structure of the normal group of rats is complete and clear, the glomerulus is uniformly distributed, and the renal tubules are orderly arranged. The kidney tissue section of the model group rat is lightly stained, the glomerular structure is not obviously pathological change, the tubular is obviously expanded and accompanied by epithelial cell flattening, and the interstitial is visibly inflammatory infiltrated. The pathological results are consistent with the serum biochemical results.
7. Analysis of results
This study investigated rat urate renal deposition models modeled by the composite inducers obtained by screening in examples 1, 2 to clarify their pathological features. The results show that the rat urate renal deposition model induced by intraperitoneal injection of 1g/kg of potassium oxazinate, 80mg/kg of adenine and 10g/kg of yeast extract in combination with 0.2mg/kg of lipopolysaccharide has the following pathological characteristics: (1) the kidney tissue is seen to have significant urate deposition, and the urate deposition of renal medulla is more pronounced than that of renal cortex. (2) Uric acid level aspect: the blood uric acid level is obviously increased, and the uric acid clearance rate is obviously reduced. (3) Aspect of renal function: the blood creatinine level is obviously increased, and the creatinine clearance rate is obviously reduced; serum β2-microglobulin levels were significantly elevated; serum cystatin C levels were unchanged. (4) Renal tissue pathological changes are mainly in the renal tubules, distention of the design lumen and flattening of epithelial cells, and inflammatory infiltration of the renal interstitium, while the glomeruli are not significantly pathologically changed. The research preliminarily defines the pathological characteristics of the rat urate renal deposition model induced by the invention, and can provide an experimental basis for the urate renal deposition pathological mechanism and the prevention and treatment research thereof.
Claims (2)
1. A method of modeling an animal model for uronate renal deposition, the method comprising the steps of:
step 1, selecting rats as experimental animals;
step 2, administering an inducer to the experimental animal, wherein the inducer consists of potassium oxazinate, adenine, yeast extract and lipopolysaccharide; the potassium oxazinate, adenine and yeast extract in the inducer are administrated by lavage, and the lipopolysaccharide is administrated by intraperitoneal injection;
wherein, the administration dosages of the potassium oxazinate, adenine, yeast extract and lipopolysaccharide in the inducer are respectively 1g/kg, 80mg/kg, 10g/kg and 0.2mg/kg of each experimental animal.
2. An inducer for an animal model of urinary acid salt renal deposition, characterized in that the inducer consists of potassium oxazinate, adenine, yeast extract and lipopolysaccharide; the administration dosages of the potassium oxazinate, adenine and yeast extract in the inducer are respectively 1g/kg, 80mg/kg and 10g/kg of each experimental animal, the lipopolysaccharide is administered by intraperitoneal injection, and the lipopolysaccharide dosage of each experimental animal is 0.2mg/kg.
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