动物造模,肝纤维化模型 zi-bio 2024-01-26 851

　　1. Animal modeling materials: Wistar rats, female, weighing 120-150g; Medications: Human serum albumin, Freund's incomplete adjuvant, prostaglandin E1.

　　2. Modeling method: Dilute human serum albumin with physiological saline, emulsify with an equal amount of incomplete Freund's adjuvant, and inject 0.5ml (containing 4mg of albumin) subcutaneously at multiple points in each rat for a total of 4 times. The interval between the first and second sessions is 14 days, and the interval between the third and fourth sessions is 10 days. 10 days after the last immunization, blood samples were taken for antibody testing. Antibody positive rats were taken for the experiment. The tail vein was attacked and injected with albumin twice a week, with 2.5mg/mouse in the first week. Afterwards, each attack increased by 0.5mg until 4.5mg/mouse, and this amount was maintained for at least 2 months. Alternatively, the first and third injections of albumin were administered at a dose of 8mg per animal, while the rest were administered at a dose of 4mg per animal, for a total of 9 times. Additionally, subcutaneous injection of prostaglandin E1200 was initiated one day prior to the injection μ G/time, twice a day, until the end of the experiment.

　　3. Modeling principle: Immune attack with human serum albumin can induce typical liver fibrosis and cirrhosis models in rats.

　　4. Pathological observation of changes after modeling: Light microscopy observation showed that in the early stage of attack injection (15 days), the liver tissue structure was generally normal. Afterwards, reticular fibers and collagen fibers proliferated in the portal area and extended outward. At the same time, it proliferates around the central vein and is scattered along the hepatic sinuses, connecting with each other. Fibrous connective tissue surrounds the lobules, which later form pseudolobules and present as typical cirrhosis. Lymphocytes, monocytes, and eosinophils infiltrate the portal area and around the central vein, exhibiting an inflammatory response. Dense staining and dissolution of small binucleated cells along the boundary plate and liver nuclei indicate mild damage and proliferation of liver cells. Under electron microscopy, active proliferation of adipocytes can be observed in the early stage, while myofibroblasts can be observed in the later stage, distributed in the portal area. There is a large amount of collagen deposition around it, and the deposited collagen forms a relatively loose separation, including functional quiescent fibroblasts.

　　Immunofluorescence staining: Positive fluorescence staining is observed in the liver sinuses, vascular walls, connective tissue, and glomeruli, indicating the deposition of immune complexes in the liver and kidney tissues.