动物造模,角膜炎动物模型 z-bio 2021-03-29 477

　　1 herpes simplex virus method

　　(1) Reproduction method (1) Select female Balb/c mice (body weight 14 to 16 g) aged 4 to 6 weeks, and inject pentobarbital sodium intraperitoneally at a dose of 45 mg/kg body weight... anesthesia Next, place it under a microscope, scratch the corneal epithelium of laboratory mice with a square 1 ml injection needle, and drop 5 μl of 2×100000000 PFU/ml virus (HSV-1RE strain) onto the scratched cornea. (2) Under general anesthesia (25 mg/kg body weight), 3% sodium pentobarbital was injected into colored rabbits weighing 2.0 to 3.0 kg through the rabbit’s ear vein and placed under an operating microscope. Take a capillary glass tube with a diameter of 1.2 mm and aspirate 1 ml of the virus strain (use HSV-I strain, incubate in Hela cells, centrifuge, and cool to -70°C for later use. Store the supernatant in In the refrigerator. Remove and rearm the virus during the inoculation process (1.15 x 100000000 PFU/ml), and close one end with plastic. The rabbit’s eyes are fixed and 25 points are inoculated on the entire cornea. During the inoculation process, glass The tube must be perpendicular to the corneal surface. According to Trefin surgery, the depth of the drill hole is limited to the penetration of the corneal epithelial layer...

　　(2) Model characteristics (1) The cornea of the model mouse on the third day of infection became less transparent than on the first day, and typical changes of dendritic lesions appeared, and the cornea became cloudy and disappeared. I achieved transparency on the 7th day of infection. (2) Severe dendritic keratitis appears on the cornea of rabbits, and severe cases develop into geographic corneal ulcers. (3) Comparative medicine Herpes simplex virus keratitis is a recurrent and persistent disease, and its incidence and blindness are the highest among corneal diseases. The model is easy to construct, has a short period, and can be copied in a large amount in a short period of time. The virus can be observed 48 hours after inoculation. High sensitivity and uniform response. Typical dendritic keratitis appeared after 48 hours. .. The model can objectively reflect the rapid elimination of the drug in the early stage, thereby inhibiting corneal inflammation, effectively, objectively and accurately determining the efficacy of the drug, and provides a means.

　　Bacteria method caused 2 eye damage

　　(1) Reproduction method The experimental rabbits weighing 2.0 to 2.5 kg were bred for 1 week and adapted, and then both eyes were examined before the experiment. When smianxin is applied to the eye of 0.2-0.3 ml per rabbit eye, the rabbit eye will show mild local anesthesia for 3-5 minutes, while pressing the cornea with trefin has no response. Using corneal aspergillins with diameters of 7 mm and 4 mm, gently push the cornea and rotate it clockwise, resulting in circular trauma. Pull the eyelid open, open it into the cup, and use a 1 ml syringe to inhale 0.1 ml of 2 x 10 12 liters/liter of bacterial liquid (Pseudomonas aeruginosa or Streptococcus pyogenes) and drip it into the rabbit’s eyes.

　　(2) Model characteristics 24 hours after bacterial inoculation, model animals may show typical symptoms of keratitis. The bacterial culture was positive on day 1, 3, and 5. Microhistopathological observations showed corneal edema, hyperemia, epithelial cell detachment, ulcers, unhealed wounds, and high inflammatory cell infiltration.

　　(3) Comparative medicine The cornea is avascular, and its nutrition comes from aqueous humor, lacrimal glands and limbal capillaries. Epithelial cells and parenchymal cells in the inflammatory stage of bacterial keratitis release proteolytic enzymes, which can deepen and expand ulcers. The conjunctiva and lacrimal gland are also involved in inflammation, showing swelling and hyperemia of the conjunctiva, lymphocytes and monocytes. Neutrophils. Tears will flush the conjunctival sac and the surface of the cornea. Tears contain antibacterial lysozyme and immunoglobulin, which have a bactericidal effect. However, the speed of body repair is closely related to the degree of corneal damage, the strength of bacterial pathogenicity, and the number of infected bacteria. The Gram-negative Pseudomonas aeruginosa and Gram-positive Streptococcus pyogenes selected for this model are both ophthalmic pathogenic bacteria, which produce a variety of decomposing enzymes, causing local tissue degradation and bacterial body inflammation. The tissue is destroyed and even perforated. The model method is simple and easy to implement, and can be used for clinical research of ophthalmic antibiotic screening.