动物造模,药效评价,缺血性急性肾衰竭 z-bio 2024-08-01 512

　　[Modeling mechanism] Removing the right kidney and clipping the left renal artery causes ischemic mitochondrial dysfunction in the kidney, resulting in the production of a large amount of hypoxanthine. Under hypoxic conditions, hypoxanthine cannot be metabolized into xanthine and accumulates in large quantities. Once blood flow is reperfusion, hypoxanthine is metabolized into purine and further metabolized into uric acid, producing a large amount of superoxide anions, which further produce hydroxyl radicals and H2O2. Through membrane lipid peroxidation, it causes damage to the kidneys, leading to cell necrosis and organ dysfunction.

　　[Method of Modeling]

　　1. Select male Sprague Dawley rats weighing 180-220g. Sodium pentobarbital (40mg/kg) was injected intraperitoneally for anesthesia, and the rat was fixed in a supine position on the operating table. After removing the hair from the abdomen, local disinfection was performed with benzalkonium bromide (Neomethoxazole). The abdominal cavity was opened longitudinally along the midline of the abdomen, and the right kidney was first exposed. The right renal pedicle and right ureter were separated under a microscope, and the entire renal pedicle and right ureter were ligated with 10 # silk thread, and the right kidney was removed. Expose the left kidney and separate the left renal pedicle. After clamping the renal pedicle with a non-invasive clamp for 60 minutes, release the clamp to restore renal perfusion. At this point, the color of the kidney changes from bright red to pale or dark red, and then to bright red, indicating successful reperfusion. After observing that there is no bleeding or oozing in the surgical field, the abdominal cavity is closed layer by layer by suturing. The anesthesia and opening of the abdominal cavity in the sham surgery group are the same as above. After exposing the left and right kidneys, blunt separation of the renal capsule is sufficient.

　　2. Renal function testing

　　Collect tail vein blood samples at 1, 6, 12, 24, and 36 hours to measure BUN and Cr concentrations. Morphological observation: 36 hours after surgery, the residual kidney was removed and fixed in a 4% neutral formaldehyde solution for 24 hours. Then, gradient ethanol dehydration was performed, followed by xylene transparency, wax immersion, embedding, continuous sectioning, and H&E staining.

　　【 Model Characteristics 】 The serum concentrations of Cr and BUN in the modeling rats began to significantly increase after 6 hours. It reached its peak within 24 hours and gradually decreased thereafter, indicating the successful establishment of an animal model of acute kidney failure. Re infusion is precise. In terms of morphology, the sham surgery group showed basic normality under light microscopy, while the ARF group showed no significant abnormalities in the glomeruli. The epithelial cells of the proximal tubules disintegrated, necrotic, and shed, and cell debris and casts were visible in the lumen. The lumen expanded, and a large amount of protein was formed in the distal tubules. Necrotic and shed cells formed cell casts, and the tissue structure was loose, with urine extravasation. The degree of tubular type and injury index of renal tubules significantly increased.

　　There are many methods for establishing animal models of acute renal failure, mainly divided into two categories: ischemic and nephrotoxic. The ischemic ARF established by excising the right kidney of rats and clipping the left renal artery has the advantages of simplicity, effectiveness, and high success rate. Moreover, the morphological and pathological changes such as renal function and renal tubules are typical and stable. It is mainly used to study the pathogenesis of ARF and the preventive and therapeutic effects of oxygen free radical scavengers on ARF.