CN-118370268-B - Construction method of uric acid crystallization kidney injury animal model

Abstract

The invention discloses a construction method of an animal model of kidney injury caused by uric acid crystals, which comprises the following steps of applying a urate crystal suspension to an experimental animal, wherein the dosage of the urate crystal suspension is 0.5-100 mg/kg. Further, the urate crystal suspension is administered to the experimental animal by injection, in particular by injection into renal cortical kidney parenchyma. The invention constructs the animal model of kidney injury caused by animal urate crystallization by using the method of kidney cortical kidney parenchyma injection, has good stability and repeatability, is simple and objective, has strong operability, reduces other possible interference factors, and provides necessary experimental foundation for researching kidney injury process caused by urate crystallization.

Inventors

• LI DELUN
• MAO WEI
• LI CHUANG
• CHEN XUESHENG
• LI YIMENG
• OuYang Jianting

Assignees

• 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院）

Dates

Publication Date

20260512

Application Date

20240429

Claims (3)

1. 1. A construction method of an animal model of kidney injury caused by uric acid crystallization is characterized by comprising the following steps of modeling an experimental animal by intrarenal injection, and applying a urate crystal suspension; The dosage of the urate crystal suspension is 50-80 mg/kg; The injection depth is 1/3-2/3 of the total width of the kidneys of the experimental animals; the experimental animal is a rat; The urate crystal suspension is sodium urate crystal suspension; The preparation of the sodium urate crystal suspension is as follows: ① 400mg of Sigma uric acid was dissolved in 80mL of purified water and heated to 60℃in a thermostatic water bath; ② Uric acid was dissolved with 3m naoh2.8ml until completely dissolved; ③ Adjusting pH to 8.9 with NaOH and glacial acetic acid, and crystallizing at room temperature for 2 days; ④ Collecting the crystals on filter paper, washing with absolute ethanol for 3 times, and drying with high fire in a microwave oven; ⑤ Collecting sodium urate MSU crystals by a centrifuge tube, and sterilizing at 180 ℃ for 2 hours; ⑥ Sodium urate MSU crystal suspension was prepared using sterile PBS.
2. 2. The method for constructing an animal model of renal injury due to uric acid crystals as defined in claim 1 wherein the suspension of uric acid salt crystals is injected by intrarenal cortical injection.
3. 3. The method for constructing an animal model of uric acid crystallization-induced kidney injury as defined in claim 1 or 2, wherein the method further comprises verifying the pathomorphology, serum uric acid level and serum creatinine level of the obtained experimental animal after the experimental animal is injected with the urate crystal suspension.

Description

Construction method of uric acid crystallization kidney injury animal model Technical Field The invention belongs to the technical field of animal model construction, and particularly relates to a construction method of an animal model with kidney injury caused by uric acid crystallization. Background Hyperuricemic Nephropathy (HN) is one of the diseases that severely threatens public health. Studies have shown a significant impact of hyperuricemia in promoting the progression of chronic kidney disease, although its underlying mechanisms remain unclear. However, researchers agree that the kidney is the main tissue responsible for uric acid excretion and reabsorption, and that uric acid rises and accumulates in the kidney following hyperuricemia to form sodium urate (MSU) crystals that cause kidney damage. However, most of animal models used in basic research of hyperuricemia at present use a method for increasing serum uric acid concentration, which is theoretically kidney damage caused by serum uric acid, and the influence of sodium urate crystals on kidneys cannot be explored. And the formation and observation of sodium urate in vivo is also one of technical problems. Therefore, the elucidation of the mechanism of kidney injury caused by sodium urate crystals is of great significance. Recent studies indicate that hyperuricemic kidney disease is kidney injury due to sodium urate crystals and not elevated blood uric acid concentrations. This is different from the animal model of the prior hyperuricemia. Therefore, it is particularly important to develop a stable sodium urate-induced kidney injury animal model. The current model research on hyperuricemia mainly comprises (1) a uric acid source increasing method, (2) uric acid excretion inhibiting method and (3) uric acid metabolism inhibiting method, which comprises the following steps: (1) The method for increasing uric acid source can directly supplement uric acid or uric acid precursor substances, mainly comprises adenine, hypoxanthine, fructose and yeast extract. The method of pouring the above drugs into the stomach is mostly used for molding in research. (2) Anti-tubercular drugs such as ethambutol and pyrazinamide can inhibit uric acid excretion to cause rise of haematuria. The method of pouring the above drugs into the stomach is mostly used for molding in research. However, ethambutol alone has little research in molding, and is often molded in combination with other chemicals. (3) The inhibition uric acid metabolism method is that the potassium oxazinate is used as a uricase inhibitor, and can inhibit the activity of uricase to highly simulate the condition of human uricase. The HUA model can be formed in a short time by single administration, but the excretion is fast, and the model cannot be formed for a long time by single use, so the model is formed by combined administration. The three models simulate the rise of blood uric acid concentration by using different modes, and the influence of the high uric acid level on the kidney is observed by prolonging the modeling time. However, all three models can not deposit sodium urate crystals in the kidney stably, and the influence of sodium urate crystals on the kidney can not be explored. Therefore, there is a need for a method of constructing and evaluating an animal model that mimics sodium urate crystals deposited in the kidney while simultaneously causing kidney damage. Disclosure of Invention The invention aims to provide a construction method of an animal model of kidney injury caused by uric acid crystallization, which can simply and stably establish the animal model of kidney injury caused by urate crystallization in kidneys. The aim of the invention can be achieved by the following technical scheme that the construction method of the uric acid crystallization kidney injury animal model comprises the following steps of applying a urate crystallization suspension to an experimental animal, wherein the dosage of the urate crystallization suspension is 0.5-100 mg/kg. Preferably, the urate crystal suspension is applied to experimental animals in an injection mode, and the dosage of the urate crystal suspension is 0.5-100 mg/kg. More preferably, the experimental animal is molded by intrarenal injection, and the urate crystal suspension is applied, wherein the dosage of the urate crystal suspension is 0.5-100 mg/kg. Specifically, the urate crystal suspension is injected by means of intrarenal injection through renal cortex. Preferably, the injection depth is 1/3-2/3 of the total kidney width of the experimental animal. Taking a rat as an example, the injection depth of the syringe needle is about 1/3-2/3 of the total width of the rat kidney, and the injection depth is about 5-10 mm, so that the tissue-pith-skin junction area of the rat kidney can be reached. Preferably, the dosage of the urate crystal suspension is 50-80 mg/kg. Preferably, the urate crystalline suspension is sodium urate crystallin