动物造模,药效评价,肾炎模 z-bio 2024-08-20 966

　　1. Modeling material animals: Male NHC rats weighing 150-200g; Drug: Isopentobarbital, complete Freund's adjuvant.

　　2. Modeling method: After anesthesia with pentobarbital, animals were euthanized and a catheter was inserted from the thoracic aorta. The portal vein was cut off and thoroughly perfused with cold saline until the kidney color turned white. Take out the kidneys on both sides, separate the renal cortex and cut it into fine pieces. Squeeze the renal cortex homogenate through a 150 mesh metal sieve, mix it thoroughly with about 4 times the amount of physiological saline, centrifuge for 10 minutes, and centrifuge the supernatant again. Take the supernatant after the second centrifugation and centrifuge at 78680 × g for 45 minutes. Discard the supernatant, mix the precipitate with distilled water, wash it three times, and centrifuge continuously for several times. After vacuum freeze-drying the last precipitate, store it in an ice box for experimental use.

　　Feed the modeling animals in metabolic cages, observe for one week before injecting antigens, and check urine protein daily. Animals without protein or pathological conditions in urine are considered qualified. Each animal was made into an emulsion of the same antigenic substance (4mg) dry weight, mixed with 0.2-0.4ml of complete Freund's adjuvant, and injected into the plantar area, only once.

　　3. Modeling principle: The same immune complex induces animal nephritis models.

　　4. Changes after modeling: During the 5th to 12th week of injection, proteinuria appeared successively, but the amount of urinary protein excretion was not high, mostly between 10-20mg/24h, and did not exceed 30mg. The duration ranged from 4-5 months to 1-2 months. At the 30th week after injection of renal antigen, there was no proteinuria in the animals.

　　Light microscopy revealed local thickening of the glomerular capillary wall, narrowing of the lumen, degeneration of the renal tubular epithelium, and no significant changes in the interstitium. Electron microscopy revealed scattered irregular swelling or nodular elevations in the middle of GBM, making it difficult to distinguish between dense and sparse layers. Occasionally, electron dense immune complexes were deposited beneath the epithelium of GBM.