动物造模,肾功能衰竭动物模型 z-bio 2021-03-26 497

　　1. Aminoglycoside antibiotics induced renal failure model

　　(1) Replication method ①Kanamycin model: The rats were deprived of water for 24 hours before the experiment, and then kanamycin was injected intramuscularly at a dose of 200 mg/kg body weight, and blood was collected for BUN and SCr 24 hours later. Take BIJN>600mg/L and SCr>15mg/L as the signs of successful model establishment. ②Gentamicin model: Gentamicin 100mg/kg body weight is injected intramuscularly every day for 7 days to complete the acute renal failure model of gentamicin nephrotoxicity. According to the scheduled time of the experiment, the animal's urine output and routine urine inspection were performed, and blood was collected for the determination of blood BUN, SCr, etc. At the same time, kidney tissue samples were taken, fixed with fixative, and routine tissue sections were made and pathological examinations under light microscope.

　　(2) Model characteristics The contents of BUN, SCr and urine N-acetyl-β-D glucosaminidase in the model rats gradually increased with the prolonged administration time. On the 4th day after the medication, the excretion of NAG enzyme and lysozyme in the urine of the model animals increased significantly, the blood urea nitrogen began to rise, and the urine protein content increased; on the 7th day, the blood urea nitrogen and creatinine increased significantly. Histopathological observations under the microscope showed that the microvilli of the brush border of the renal tubules were necrotic, shedding and fused, there were cell fragments in the lumen, the renal tubule epithelial cells were degeneration and necrosis, and the renal interstitium could be seen in the renal interstitium, mainly inflammatory cells infiltration. . The damage of renal proximal tubule epithelial cells can be aggravated with the time of medication.

　　(3) Comparative medicine Gentamicin (GM) and kanamycin are both aminoglycoside antibiotics (AG), and the pathogenesis of ARF in animals is basically the same. When aminoglycoside antibiotics enter the animal body, they can be reabsorbed in the renal tubules and accumulated in the lysosomes of renal tubular epithelial cells. A certain dose will inhibit phospholipase A and C in the lysosomes of renal cortex cells, leading to phosphatidylinositol It accumulates in the lysosome, causing the lysosome to swell, rupture, release various proteolytic enzymes, cause other subcellular membrane damage and renal tubular necrosis, and eventually form acute renal failure. This model can be used in clinical experimental research on the prevention and treatment of ARF caused by aminoglycoside antibiotics.

　　2. Mercury chloride induced renal failure model

　　(1) Reproduction method Adult New Zealand rabbits weighing about 2kg, HgCl2 is freshly prepared with normal saline to a 0.2% concentration solution, and injected into both hind limbs of rabbits at a dose of 5-15 mg/kg body weight for 2 days. It can be prepared into an acute renal failure (ARF) model. In this method, rats with a body weight of about 200 g and mice with a body weight of about 20 g can also be used as model animals, and the HgCl2 injection dose is the same. Blood samples were collected for blood biochemical determinations at the predetermined time in the experiment, and kidney tissue samples were taken, fixed in the fixative, and routinely sliced and examined under a light microscope.

　　(2) Model characteristics: On the second day after HgCl2 injection, the renal blood flow of the model animals decreased, and the SCr level rose sharply to above 176.8 μmol/L and continued for many days; the BUN level could rise to 5304 on the third day after injection ~6188/μmol/L or more; 24h myocardial clearance rate is significantly reduced; 24h myocardial liver and total urea nitrogen excretion are significantly reduced. Animals have obvious oliguria within 3 to 5 days after HgCl2 injection, followed by polyuria; protein and various casts can appear in the urine, with the most obvious being 3 to 5 days after injection. Macroscopic pathological examination of the model animals’ kidneys was significantly enlarged, weight increased (increased renal index), renal cortex ischemia, and medulla congestion. The histopathological observation under the microscope showed that it was mainly the degeneration and necrosis of the proximal convoluted tubule of the kidney. When low-dose mercury is poisoned, only the necrosis of the straight epithelial cells of the proximal convoluted tubule of the kidney is shown; when high-dose mercury is poisoned, necrosis of other segments of the proximal convoluted tubule may also occur. The model replication method is simple and easy to implement, and the lesions are relatively stable.

　　(3) Comparative medicine The model-building mechanism of this model is that Hg2+ can be reabsorbed by the renal tubules after being filtered by the glomerulus, and accumulated in the cells, combining with the sulfhydryl (SH) and disulfide groups in the renal tubular epithelial cells , Affects the activity of cells and intracellular enzymes; affected cells damage the function and structure of cells and their membranes due to mercury-sulfur reaction, loss of cell respiration, leading to cell degeneration and necrosis, part of it falls off the lumen and blocks the renal tubules, causing original urine Through obstruction, the permeability of the necrotic renal tubules increases, the original urine oozes out and returns to the renal blood vessels, and the body produces clinical manifestations such as oliguria and anuria. At the same time, the original urine leaks to the renal interstitium, which can form interstitial edema, which further compresses the renal tubules, causing acute renal failure in the animal. HgCl2 poisoning can also cause the redistribution of renal blood flow in the animal body, causing renal cortex ischemia, medulla congestion, and decreased renal tubular filtration, which is also one of the causes of ARF.

　　3. Snake venom-induced renal failure model

　　(1) Replication method A male adult rat weighing about 300g is injected with agkistroxin at a dose of 0.4mg/kg body weight through the vein to prepare an acute renal failure model. Blood samples were collected for blood biochemical determinations at the predetermined time in the experiment, and kidney tissue samples were taken, fixed in the fixative, and routinely sliced and examined under a light microscope.

　　(2) Characteristics of the model After intravenous injection of snake venom, the urine output and GFR of the model rats decreased rapidly, and the urinary sodium excretion fraction also decreased significantly. After 48 hours, the GFR was still lower than before the injection, but the urine output increased slightly. Microscopic histopathological observation showed that necrosis of renal distal convoluted tubules and collecting tubules appeared, manifested by the shedding of tubule epithelial cells, nucleus dissolution and disappearance; glomerular capillary loop segmental necrosis, massive cellulose deposition and neutrophils Cell infiltration. After 48 hours of injection, the morphology of the glomerulus can return to normal, but the necrosis of the renal tubules is still clearly visible. A large number of red blood cell casts are still visible in the lumen.

　　(3) Comparative medicine. The model-building mechanism of this model is that snake venom can cause direct damage to the muscles of the model animal, leading to massive hemoglobinuria in the body, causing obstruction of the renal tubules; at the same time, snake venom can activate factor X and thrombin in the body, causing blood vessels. Internal coagulation and thrombosis can destroy the kidney microcirculation; snake venom can also directly damage the red blood cell membrane, leading to intravascular hemolysis and the formation of large amounts of hemoglobinuria, which ultimately leads to damage to the kidney function and morphology of model animals, and acute renal failure. This model can be used for clinical research on the pathogenesis and drug treatment of human ARF caused by snake venom.

　　4. Adriamycin-induced renal failure model

　　(1) Reproduction method Adult rats weighing about 200g are injected with adriamycin (ADR) at a dose of 6mg/kg body weight once through the tail vein of the rat. Urine was collected at the predetermined time for the experiment, and blood was collected for the determination of blood biochemical indicators. After the animals were sacrificed, the kidney specimens were taken and fixed in a fixative solution for routine tissue sectioning and observation under a light microscope. And make electron microscope specimens for observation by transmission electron microscope.

　　(2) Model characteristics Model animals can produce a large amount of proteinuria on the 7th day after injection, and the proteinuria is more obvious on the 14th day. At the same time, a large amount of ascites, hypoproteinemia and hypercholesterolemia can occur. Microscopic histopathological observation of renal tubules and glomeruli can have slight pathological changes. The ultrastructure under the electron microscope showed that the capillary endothelial cells and podocytes were swollen, and the foot processes were fused and flattened.

　　(3) Comparative Medicine Adriamycin (ADR) is a kind of anti-tumor drug with quinoid structure commonly used in clinical practice. One-time injection of doxorubicin can induce ARF model, and its pathogenesis is mainly related to the body's oxygen free radicals and lipid peroxidation damage. Under the action of drug-metabolizing enzymes in the body, ADR forms semiquinone free radicals. Semiquinone free radicals react with oxidation to generate superoxide anions and hydroxyl free radicals, both of which can cause damage to the kidneys through the lipid peroxidation of the membrane. This model is similar to the minimal change type of human glomerular disease.

　　5. Sodium fluoride induced renal failure model

　　(1) Reproduction method Adult rats weighing about 200g are given a one-time oral dose of sodium fluoride at a dose of 135mg/kg body weight to prepare an ARF model. Observe the acute nephrotoxicity induced by sodium fluoride and determine the relevant urine biochemical indicators.

　　(2) The characteristics of the model can clearly observe the acute nephrotoxicity induced by sodium fluoride in rats. With the prolonged observation time, the urine output of the model animals increases significantly, and the urine fluoride, a-glutathione-S-transferase (a-GST), N-acetyl-β-D-glucose (NAG) and creatinine concentrate content all changed.

　　(3) Comparative medicine The amount of fluoride given to animals during the modeling of this model largely affects the above urine biochemical indicators. The degree of damage of fluoride to kidney tissue is that a-GST can show a strong and lasting change, so it can be inferred that the content of a-GST can be used as an important indicator of the degree of proximal convoluted tubule damage. This indicator is of great significance for early detection and long-term observation of renal proximal tubule damage caused by human fluoride poisoning in clinical practice. This model helps to formulate guidelines for the treatment of renal failure caused by human fluorosis.

　　6. Cisplatin-induced renal failure model

　　(1) Reproduction method Adult rats weighing about 200g are injected with cisplatin at a dose of 6mg/kg body weight through the tail vein of the rat, or cisplatin 10mg/kg body weight is dissolved in 1ml saline for a single intraperitoneal injection . After the injection, blood was collected from the tail vein at a predetermined time for blood SCr and BUN determination. The animals were anesthetized on the 6th and 10th days after the injection, and their kidney specimens were fixed in 10% neutral formaldehyde solution and embedded in paraffin. Regular tissue sections were made, and the sections were stained with HE and pathological observation under light microscope.

　　(2) Model characteristics After injection, the blood SCr and BUN of model rats were significantly increased. Under the microscope, renal histopathology observed necrosis of most of the tubules outside the renal medulla, necrosis and shedding of tubule epithelial cells, and pyknosis, dissolution and disappearance of nuclei. The lumen of the tubule is expanded, and there are falling debris in the cavity. The pathological changes of kidney tissue were basically similar on the 6th and 10th day after injection.

　　(3) Comparative medicine Cisplatin (CP) has certain toxicity to human body in clinical application, and its incidence exceeds 20%. This model uses the toxic effects of cisplatin on the body’s kidneys to replicate the clinical manifestations of ARF disease. The model replicated by this method can be used to evaluate the prevention and treatment effects of L-arginine, glycine, atrial natriuretic peptide combined with dopamine on ARF Research.

　　So far, clinical research on the pathophysiology and pathogenesis of ARF has made positive progress, and making an ideal animal model of ARF for experimental therapeutic research is the prerequisite and basis for clinical therapeutic research. At present, there are many ARF animal models made based on the cause of ARF. These models provide good research ideas for the study of the pathogenesis, development rules and biological characteristics of ARF; for the study of ARF pathogenesis, therapy and pharmacodynamics, It has laid an important experimental foundation; it provides an important research carrier for seeking and understanding the target of action, efficacy, and action links and mechanisms of ARF drugs. Due to the use of model animals and replication methods, currently commonly used ARF animal models have their own characteristics. Researchers should select appropriate animal models to carry out research according to the purpose of the research.