动物造模,药效评价,口腔溃疡 z-bio 2024-08-27 544

　　(1) The replication method first involves placing the dialysis bag in a 0.5mol/L EDTA solution, boiling for 0.5 hours, discarding the solution, replacing it with distilled water and boiling it three times. Then, it is stored in distilled water containing 0.01% sodium azide and stored at 4 ℃ for future use. New Zealand rabbits weighing 1.5-2.0kg were euthanized using anesthesia, and their oral mucosal tissue was removed. The homogenizer was used to prepare the homogenate, which was then shaken with ultrasound and centrifuged at low temperature to retain the precipitate. The supernatant was then dialyzed to filter out small molecule substances, and the large molecule protein was retained in a dialysis bag. The dialysis process was repeated 5 times using the same method, and finally the contents of the bag were freeze-dried to obtain powdered protein as the antigen, which was stored in a -70 ℃ freezer for later use. Take New Zealand rabbits weighing 1.5-2.0kg, dissolve 12.5mg of pre extracted oral mucosal protein in 0.5ml of phosphate buffer solution, and add 0.5ml of complete Freund's adjuvant to form an emulsion. Inject 1ml of the above tissue emulsion intradermally on both sides of the rabbit spine, for a total of 20 injections. Inject once every 2 weeks using the same method, for a total of 5 injections. Cut the oral mucosa at different times for routine pathological tissue sectioning and light microscopy examination.

　　(2) The characteristics of the model: In the early stage of modeling, local mucosal congestion began in the model rabbits, followed by ulceration. From the 8th day after the second injection of antigen, ulceration could occur in the oral mucosa of the animals. Ulcer lesions often occur on the upper and lower lips of animals, followed by the gums and oral mucosa. The diameter of the ulcer is 1-5mm, mainly circular or elliptical in shape, with relatively neat edges, and its surface is covered with a layer of yellow pseudomembrane; During the model making process, ulcers usually heal on their own within 2-3 days and then recur. Microscopic histopathological observation of oral mucosa showed necrosis and shedding of mucosal epithelium, with a large amount of neutrophils and cellulose like exudate covering the ulcer surface. Small blood vessels at the bottom of the ulcer were dilated and congested, accompanied by infiltration of neutrophils, lymphocytes, and monocytes.

　　(3) Recurrent Aphthous Ulcer (RAU) is the most common oral mucosal disease in clinical practice, and its etiology is still unclear. In recent years, most research reports have suggested that it is related to the body's immunity, especially autoimmunity. Therefore, establishing an ideal and suitable animal model for recurrent oral ulcers (RAU) is of great significance. Due to limitations in people's understanding of the etiology of RAU, there are currently no specific drugs or methods for treating RAU in clinical practice. The model replicated using immunological methods has disease onset characteristics, oral mucosal pathological damage, and is extremely similar to human RAU in clinical practice, with non-specific inflammation observed under the microscope. The immunological RAU model established by this method may be produced by the specific protein acting as an antigen entering the body, producing specific antibodies and inducing oral mucosal immune responses in animals. This model can be used for experimental research on the pathogenesis and drug treatment of human RAU.