动物造模,肾小管间质纤维化模型 z-bio 2021-03-26 323

　　(1) Method of reproduction The anesthesia method of sodium pentobarbital is to inject 30 mg/kg of body weight intravenously into a New Zealand rabbit weighing 2 to 3 kg, fix it on its back operating table, and regularly disinfect the abdominal skin. And hair loss. Make a straight incision on the outer side of the peritoneal rectal muscle below the ureter, make an incision on the lateral oblique muscle aneurysm, expose the medial oblique muscle, separate the ventrolateral side, push the peritoneum inward, and separate the peritoneum straightforwardly, releasing the ureter in front of the bone vessel and releasing ureter. Fix the ureter with Allis forceps, cut one third of the diameter of the ureter (4-5 cm from the renal pelvis here) with scissors, and then insert the F2 catheter retrogradely 2-3 cm. Urine becomes visible at the proximal end of the ureter. After the catheter is emptied, the distal end of the ureter is ligated with #1 silk thread, the ureter and the catheter are fixed, and the distal end of the catheter is ligated and sealed. It is buried under the skin for specimen collection. At a given time, the model rabbits were injected with 3% inulin solution intravenously at a dose of 120-150 mg/kg body weight. Next, the obstructive renal pelvis, non-obstructive renal pelvis and blood samples were taken to determine the level of inulin and transforming growth factor β1 (TGF-β1) and the animals were sacrificed. A kidney specimen was taken and fixed with 10% formaldehyde. Normal paraffin tissue sections, HE stained, and observed with an optical microscope.

　　(2) Model characteristics After surgery, the concentration of inulin in the urine of the obstructive side of the model animals was significantly lower than the normal value, but the concentration of TGF-β1 in the urine of the obstructive side was significantly lower than the normal value. It is higher than the normal side, the renal function of the obstructed side is decreased, and the secretion of TGF-β1 is significantly increased. It is observed that the total occluded kidney volume of the model animals is larger than that of normal people, the renal pelvis and renal pelvis are enlarged, the ureter is enlarged, and the lumen is enlarged. Histopathological observation under the microscope showed that the renal cortex of the occluded kidney was congested and edema, with different amounts of inflammatory cell infiltration; renal interstitial congestion and edema; different degrees of inflammatory cell infiltration; small morphological glomeruli and renal edema were basically Normal, no obvious changes. In the early stage of obstruction in this model, kidney diseases are mainly inflammation of the renal interstitium, congestion, and edema. In the middle and late stages, renal interstitial fibrosis and renal insufficiency may occur. The glomeruli gradually shrink and replace thin glomeruli. organization. The changes of inulin levels in urine of occluded kidneys of model animals can accurately reflect the pathological changes of glomerular filtration rate reduction, and the continuous excess of TGF-β1 may occur during the formation of renal fibrosis. The model method is simple, practical, easy to collect repeated specimens, and can be used for dynamic research. In the process of creating this model, the length of catheter insertion must be controlled between 2 cm and 3 cm to avoid damage to the renal pelvic mucosa and renal parenchyma due to over-insertion, renal pelvis and renal pelvis bleeding or infection.

　　(3) The characteristic of a comparative medical unilateral ureteral obstruction (UUO) nephropathy model is progressive tubulointerstitial fibrosis with glomerular disease, but no hypertension or dyslipidemia. Clinically, unilateral ureteral obstruction can be caused by a variety of diseases. The continuous attack of the disease can cause different degrees of renal insufficiency, until the complete loss of renal function. The direct cause of glomerular and tubular interstitial fibrosis is a variety of extracellular matrix, such as fibronectin (FN), type I, type III and type IV collagen, epidermal sulphate proteoglycan (Biglycan) and others Proteoglycans. The changes of a series of growth factors and cytokines are closely related to the synthesis and metabolism of these ECM components. Transforming growth factor β1 (TGF-β1) is one of the most important cytokines. TGF-β1 stimulates the cell synthesis of collagen, fibrin, laminin and proteoglycan through receptor signaling, and regulates the damage and repair of normal tissues. Its overproduction is related to chronic fibrosis. This model is improved on the basis of the traditional unilateral ureteral ligation method, and uses a built-in ureter to prepare for unilateral ureteral obstruction. In addition, the same obstructed animal model created by conventional surgical methods can be obtained, and it has the typical characteristics of kidney injury and fibrotic lesions. It has been confirmed that model animals may have reduced inulin levels and increased TGF-β1 levels. It can be used for dynamic and reproducible research on the etiology and drug treatment of renal interstitial fibrosis.