动物造模,缺血性急性肾功能衰竭动物模型 z-bio 2021-03-26 247

　　1. One side renal ischemia/reperfusion plus contralateral nephrectomy model

　　(1) Reproduction method ① Male rats weighing 200～300g were anesthetized by injecting pentobarbital sodium at a dose of 30 mg/kg through the abdominal cavity. After anesthesia, the animals were fixed on the operating board on their back, and the abdominal operating area was routinely disinfected and depilated. , Make a midline abdominal incision, perform right nephrectomy, bluntly separate the left renal capsule, block the left renal artery with a non-damaged vascular clip for 60 minutes, loosen the clip to restore renal perfusion, observe before reperfusion, 15min, 24h after reperfusion Changes in the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in kidney tissue. ②Male rats weighing 200～300g were anesthetized by injecting pentobarbital sodium at a dose of 30 mg/kg through the abdominal cavity. After anesthesia, the animals were fixed on the operating board on their back, and the abdominal operating area was routinely disinfected and depilated, and a midline abdominal incision was made. , Bluntly separate the capsules of both kidneys, block both renal arteries with non-traumatic arterial clamps for 60 minutes, and then loosen the clamps to restore renal perfusion to prepare an ARF model.

　　(2) Model characteristics After renal ischemia/reperfusion, the SOD content in the kidney tissue of the model rats can be decreased, and the MDA content will increase significantly, and the content of both will show more obvious gradual changes with the reperfusion time. The modeling mechanism of this method is based on the mitochondrial dysfunction during renal ischemia, ATP cannot be used, and a large amount of hypoxanthine is degraded in turn. Under hypoxic conditions, hypoxanthine cannot be metabolized into purines in the body and accumulate in large amounts; at the same time, renal xanthine dehydrogenase will rapidly transform into xanthine oxidase during ischemia. When the blood is reperfused, under aerobic conditions, yellow Purine oxidase can metabolize accumulated hypoxanthine into purine, and then further produce uric acid. During this process, the body will produce a large amount of superoxide anions, and further generate hydroxyl free radicals (OH·) and H2O2. These free radicals can cause damage to the kidney tissue through the lipid peroxidation of the membrane; that is, the biological membrane ( For example, a large number of unsaturated fatty acids contained in cell membranes are degraded to produce cytotoxic lipid peroxides, causing cell necrosis and decreased organ function. Lipid peroxide will eventually be broken down into many end products including MDA.

　　(3) Comparative Medicine Clinically, ARF is an acute kidney disease characterized by a rapid change in renal function within a few hours to a few days, leading to the ability of the kidneys to excrete nitrogenous metabolic wastes, and the loss of the function of maintaining water and electrolyte stability in the body. The kidney is an organ with sufficient blood supply in the body and is extremely sensitive to ischemia. Acute tubular necrosis and renal failure can occur after 1 day of renal ischemia. However, after short-term ischemia and reperfusion, the kidney can repair the damaged tubules through the rapid proliferation of tubule epithelial cells, and renal function will be restored. This model uses surgical methods to block the blood supply of the model animal’s kidneys, causing renal failure to function, leading to hypoxia and ischemia of renal parenchymal cells, and ultimately causing renal tubular epithelial cell damage, and affecting the body’s ion exchange, reabsorption and excretion Function, clinical manifestations of acute renal failure. The ARF model established by this method can be used to investigate the pathogenesis of ARF and the study of the effects of oxygen free radical scavengers on the prevention and treatment of ARF.

　　2. Partially ligated abdominal aorta model

　　(1) Replication method Rats weighing 200～300g are anesthetized by injecting pentobarbital sodium at a dose of 30mg/kg body weight into the abdominal cavity. After anesthesia, the animals are fixed on their backs, and the abdominal operation area is routinely disinfected and depilated. The groin skin and muscle were bluntly separated from the right femoral artery, and a PE-50 catheter was inserted into the artery to monitor blood pressure. Then make a midline abdominal incision, perform right nephrectomy, separate and expose the superior mesenteric artery and left renal artery, pass a suture under the abdominal aorta in between, close the two ends and rotate in the same direction to gradually compress the abdomen The aorta, until the blood pressure of 217～313kPa measured through the femoral artery cannulation, put the kidney in hypoperfusion for 1h, relieve the compression, and suture the abdominal wall by routine surgery. 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 The blood creatinine values of the model rats were significantly increased at 24, 48, and 72 hours after the operation, and the kidneys were swollen on the naked eye. Histopathological observation under the microscope showed diffuse swelling of renal tubular epithelial cells, a small amount of small renal tubular necrosis, casts in the lumen, and obvious edema of the renal interstitium.

　　(3) Comparative Medicine Clinically, acute renal failure (ARF) is mostly secondary to the continuous hypoperfusion of the kidney. However, the pathogenesis of ARF replicated by the traditional renal artery clipping method is different from the ARF caused by continuous hypoperfusion. Compared with the renal artery clipping model, the hypoperfusion ARF model established by this method has more important significance in studying the protective effect of reduced glutathione on the kidneys of ARF, and it is more important than the actual situation of ARF patients in clinical practice. similar.

　　3. Hemorrhagic shock renal failure model

　　(1) Reproduction method A Beagle dog weighing about 10kg is anesthetized by injecting pentobarbital sodium at a dose of 30mg/kg body weight through the abdominal cavity. After anesthesia, the animal lies supine and fixed on the operating table, and the right groin is routinely disinfected and depilated. Surgical incision, blunt separation of the right femoral artery, intravenous injection at a dose of 500 U/kg body weight, heparinized canine blood, intubated through the right femoral artery, and connected to a blood storage tank with a pressure of 5.3 ~ 6.7 kPa to form an open circulatory system . The temperature of the blood storage tank is maintained at 37°C, and oxygen is continuously supplied and stirred for anticoagulation. After bleeding, the average arterial pressure of the model animals was 5.3～6.7kPa for 150min. The blood was returned to the animal within 15 minutes after the end of bloodletting.

　　(2) Features of the model This model method rebuilds the external blood circulation system of the kidney locally, so that the model animal body loses kidney function, and at the same time causes kidney ischemia and hypoxia, leading to symptoms of acute renal failure in the body, and damage to kidney tissue cells And irreversible necrosis occurred.

　　(3) Comparative medicine The model-building mechanism of this model is similar to clinical acute renal failure caused by hemorrhagic shock. Therefore, although the model-making method is complicated, it is still often used in the research of ARF drug screening.

　　4. Glycerin-induced renal failure model

　　(1) Reproduction method Adult rats weighing about 200g were deprived of water for 16 hours, and then injected 50% glycerol normal saline into the muscles of both hindlimbs at a dose of 100ml/kg body weight. In order to facilitate multiple blood collections, New Zealand rabbits weighing about 2 kg can also be used to inject 50% glycerol normal saline into both hind limbs and thighs at a dose of 15 ml/kg body weight. Blood was collected according to the scheduled time of the experiment for determination of blood creatinine and urea nitrogen, and kidney tissue specimens were taken, fixed with fixative, and routine tissue sectioning, pathological examination under light microscope.

　　(2) Model characteristics: 2 hours after the injection of glycerol, the model animals can develop grape-red hemoglobinuria. After 24 hours, blood urea nitrogen will increase significantly, and there will be oliguria or anuria. Microscopic histopathological observations showed that the glomeruli had mild hemorrhage; extensive granular degeneration of the renal tubules, necrosis of some tubular epithelial cells, and lumen occlusion; a large number of hyaline casts were seen in the cavity, mild hemorrhage of the renal tubules, and renal interstitium Edema and infiltration of inflammatory cells; renal vascular congestion and so on. Model animals can have different degrees of acute renal failure within 1 to 3 days after administration. The modeling mechanism of this model is mainly that the injection of glycerol can cause muscle dissolution and hemolysis of the body, release a large amount of hemoglobin and myoglobin, and myoglobin can cause renal vasoconstriction, and the small arteries of the glomerulus entering the glomerulus will contract, causing renal blood. The flow rate and glomerular filtration rate decrease, causing renal ischemia and causing damage to renal tubular function. Because the renal tubules cannot be reabsorbed, they form a cast to block the renal tubules, causing pathological damage to the renal tubules and interstitium. At the same time, both myoglobin and hemoglobin can be decomposed into methemoglobin, which has a direct toxic effect on the renal tubules.

　　(3) Comparative medicine The hemoglobinuria ARF model established by this method is very similar to the ARF caused by severe tissue trauma in clinic, and it is squeezing. An ideal animal model for renal failure, the animal used in this model is cheap and easy to obtain, the method is simple, and the disease is stable. If you need to make dynamic observations within a certain period of time, rabbits can also be used for multiple blood sampling. It has good application value in clinical research on the pathogenesis of human traumatic ARF, drug prevention and treatment effects, drug screening, and new drug evaluation. .

　　5. Norepinephrine renal failure model

　　(1) Reproduction method Beagle dogs weighing about 10kg are anesthetized by injecting pentobarbital sodium at a dose of 30mg/kg body weight through the abdominal cavity. After anesthesia, the animal lies supine and fixed on the operating table. The abdominal operation area is routinely disinfected and dehaired. A midline abdominal incision was made, one side of the ureter was surgically separated, and a cannula was inserted to collect urine; the renal artery was separated, norepinephrine was instilled through the renal artery at a dose of 0.75 μg/kg body weight per minute, continuous infusion for 40 minutes, and the abdominal wall was sutured by routine surgery . The urine volume was determined according to the predetermined time of the experiment, and kidney tissue specimens were taken, fixed in the fixative, and routinely sliced for pathological examination under light microscope.

　　(2) Model features After treatment, model animals have a series of renal failure symptoms such as a sharp decrease in renal blood flow and a decrease in the urine output of the ipsilateral ureter. 2 hours after treatment, it can cause severe renal tissue ischemia and renal tubular necrosis, and may appear severe Of microcirculation disorders. Histopathological observations under the microscope showed that after 40 minutes of medication, necrosis of the proximal renal tubules was seen, and foam-like substances and casts were seen in the distal tubules and collecting ducts, but there was no obvious change in the glomerulus; after 2 hours of medication, the proximal tubules of the kidney could be seen. Extensive necrosis occurred, and renal tubular epithelial cells were obviously flattened. This model has a stable course of disease, high success rate, and continuous blood sampling for testing.

　　(3) Comparative medicine The ARF model established by norepinephrine is basically similar in pathogenesis to the ARF model caused by renal artery ischemia/reperfusion, and both belong to ischemic ARF. Injecting norepinephrine can cause renal vasoconstriction in model animals, causing a decrease in glomerular blood flow and glomerular filtration; renal tissue ischemia and hypoxia aggravate renal tissue damage, resulting in ARF. The disease characteristics and course of this model are stable, the model has a high success rate, and it is convenient to take blood. It can continuously take blood for testing, and dynamically observe the time and characteristics of drug action. It can be used for clinical drug treatment and drug screening for acute renal failure. Aspects of experimental research.

　　6. Oleic acid-induced renal failure model

　　(1) Reproduction method Adult rats weighing 200 g are anesthetized by injecting pentobarbital sodium at a dose of 30 mg/kg body weight through the abdominal cavity. After anesthesia, the animals are fixed on the operating board on their back, and the abdominal operating area is routinely disinfected and depilated. A midline incision was made in the abdomen, one side of the renal artery was separated, and oleic acid was injected through the renal artery at a dose of 0.1ml/kg body weight; the abdominal wall was sutured by routine surgery. 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 the injection of oleic acid, the blood BUN of the model animals was significantly increased. Microscopic histopathological observation showed that the glomeruli and the capillary endothelial cells around the renal tubules appeared swelling, necrosis, and renal tubular necrosis. The degree of lesion damage of the kidney tissue structure of model animals can be aggravated with the increase of the dose of oleic acid.

　　(3) Comparative medicine The ARF animal model established by injection of oleic acid is characterized by severe renal microcirculation disorders, combined with renal interstitial and parenchymal damage. This model helps to understand the pathogenesis of ARF more comprehensively, and can be used to simulate human ischemic ARF originating from renal microcirculation disorders.