[Animal Experiment]-Animal Model of Ischemic Acute Renal Failure
动物实验,肾衰竭
z-bo
2022-02-03
486
[Model mechanism] Removal of the right kidney and clamping of the left renal artery will cause renal ischemic mitochondrial dysfunction and produce large amounts of hypoxanthine. In the hypoxic state, hypoxanthine cannot be metabolized into xanthine and accumulate in large amounts. When the blood is reperfused, hypoxanthine is metabolized into purine, and then metabolized into uric acid, producing large amounts of superoxide anions, hydroxyl radicals and hydrogen peroxide. Membrane lipid peroxidation can damage the kidneys, leading to cell necrosis and impaired organ function.
[Modeling method]
1. Choose a male Sprague Dawley rat weighing 180-220 grams. Intraperitoneal injection of sodium pentobarbital (40 mg/kg) anesthetized, fixed on the operating table in the supine position, hair removal from the abdomen, then local disinfection with benzalkonium bromide (ceramide), and a midline incision was made. Open along the abdominal cavity. Expose the right kidney, separate the right renal pedicle and right ureter under a microscope, ligate the entire renal stem and right ureter with 10# silk thread, and remove the right kidney. Expose the left kidney and separate the left kidney pedicle. 60 minutes after the renal pedicle was clamped with noninvasive forceps, the forceps were released to restore renal perfusion. At this time, the color of the kidney first changes from bright red to light red or dark red, and then to bright red, indicating normal reperfusion. After bleeding or no bleeding was observed in the surgical area, the abdominal cavity was closed once with sutures. In the sham operation group, the methods of anesthesia and abdominal opening were the same. After the left and right kidneys were exposed, the kidney capsule was bluntly separated.
2, 1, 6, 12, 24, and 36 hours later, tail vein blood will be collected for renal function tests to determine BUN and Cr concentrations. Morphological observation: 36 hours after the operation, the remaining kidney was taken out and placed in 4% neutral formaldehyde solution for 24 hours. After 24 hours, the gradient ethanol was dehydrated, the xylene was clarified, waxed, embedded, and serial sections were prepared and H&E stained. [Model Features] The serum Cr and BUN concentrations of model rats began to increase significantly after 6 hours. After reaching a peak in 24 hours, it gradually decreased, indicating that an animal model of acute renal failure has been established. Reperfusion is accurate. Morphologically, the sham operation group was basically normal under an optical microscope, and the glomeruli in the ARF group had no obvious abnormalities. The proximal tubular epithelial cells are destroyed, necrotic and shedding. Cell debris and casts can be seen in the lumen. The lumen is expanded and distal. A large amount of protein is formed in the terminal tubules, the dead skin cells form cell clusters, the histology is loose, and the urine is discharged from the blood vessels. The degree of renal tubule casting and the index of damage increased significantly. [Model Evaluation and Application] The methods of establishing acute renal failure animal models can be divided into two categories: ischemic and nephrotoxic. Ischemic ARF is established by removing the right kidney of the rat and clamping the left renal artery. It has the advantages of simplicity, effectiveness, high success rate, shape and renal tubules such as renal tubules. The pathological changes are typical and stable. It is mainly used to study the cause of ARF. The effect of oxygen free radical scavenger on ARF and its preventive and therapeutic effects.
