动物造模,药效评价,肾小管间质纤维化 z-bio 2024-07-08 499

　　【 Modeling mechanism 】 Ligation of renal veins leads to renal ischemia and tubular injury, resulting in ischemic atrophy and tubular disappearance, renal interstitial hyperplasia, and renal tubulointerstitial fibrosis.

　　【 Modeling Method 】 Wistar rats weighing 200-250g were selected. Intraperitoneal injection of 8.5% chloral hydrate (0.1ml/kg) for anesthesia. After local shaving, disinfect the area with a perforated cloth, cut the skin along the midline of the abdomen to the abdominal cavity, separate the left renal vein, and ligate it with a 4-0 surgical suture. Rats were euthanized on the 5th, 10th, 15th, 20th, and 25th day after surgery, with a focus on observing the ligated kidneys. Before execution, place the rat in a metabolic cage and leave urine for 24 hours for routine urine testing. Collect femoral artery blood to detect Cr content. Bilateral kidneys were fixed in 10% neutral formaldehyde solution, routinely dehydrated, embedded in paraffin, and sliced into 4 μ m thick sections for H&E, PAS, PASM, and Masson staining, respectively.

　　【 Model Features 】

　　1. The typical lesion is stable, and the lesion on the surgical side of the kidney is highly similar to that of human TIF. Through dynamic observation, the early histological changes of the lesion are mainly characterized by degeneration and atrophy of renal tubular epithelium, interstitial congestion, and edema. Mononuclear cell infiltration and type III collagen proliferation can be seen in the interstitium. Atrophy and disappearance of renal tubules in the middle and late stages, thickening of arterial walls, increased number of renal interstitial fibroblasts, collagen deposition, and type I collagen proliferation. Throughout the course of the disease, ischemic changes can be observed in the glomeruli.

　　2. Short cycle, simple operation method, only need to separate renal vein and ligate, the surgical process only takes 20 minutes. Unilateral kidney surgery does not damage the contralateral kidney, so it has a slight impact on the animal's life. The experimental period is short, and ideal renal tubulointerstitial fibrosis lesions can be formed on the 10th day after surgery.

　　【 Model Evaluation and Application 】 Renal vein ligation modeling method is a simple, high success rate, stable lesion, and highly similar animal model of renal tubulointerstitial fibrosis to human lesions. This model overcomes the shortcomings of other models, including common methods such as unilateral ureteral ligation, subtotal nephrectomy, immunomediation, gentamicin induced renal tubular injury, adenine induced fibrosis, radiation injury, infection, etc. Each method has its own shortcomings. Drug induced methods such as cyclosporine nephritis and doxorubicin nephritis, although simple and highly successful, have high costs, long experimental cycles, and mild interstitial changes. Subtotal nephrectomy can cause TIF lesions, but the operation is complex and can cause significant trauma to animals. The immune mediated nephritis model can obtain TIF lesions by prolonging the pathogenic cycle. But the results obtained are unstable. Although the pathological changes in the adenine induced fibrosis model are stable, it requires daily administration of drugs according to the animal's weight, which is cumbersome and time-consuming. The model of renal tubular injury induced by gentamicin is mainly characterized by acute necrosis and regeneration of renal tubular epithelium, with no significant interstitial fibrosis. In the unilateral ureteral ligation model, renal tissue was injured by urine retention and compression, leading to interstitial fibrosis. The time was generally controlled at about 15 days. If the time was prolonged, the renal parenchyma became too thin due to compression. By the 15th day, interstitial fibrosis was not obvious.