动物模型,SLE z-bo 2020-08-03 645

　　[Modeling mechanism] Immunize rabbits with allogeneic hepatocyte protein, and induce various autoantibodies, abnormal renal function and proteinuria. It is similar to the clinical manifestations of human SLE.

　　[Modeling method] Aseptically separate liver tissues of the same type of rabbits, add PBS, homogenize and remove the supernatant, centrifuge at 500 xg for 3 minutes at 4°C, and pre-cool with protease inhibitors. Add buffer to the pellet. Vortex vigorously for 15 seconds at maximum speed, place on ice for 15 minutes, vortex vigorously twice for 5 seconds, and centrifuge at 16000 xg for 5 minutes at 4°C to obtain total cytoplasmic protein.

　　New Zealand female rabbits, 500 μg (final concentration 1 mg/ml) allogeneic rabbit hepatic cytoplasmic protein in the ear veins of the 0 day and 1, 2, 4, 6, 8, 10 and 12 weekends have been injected. Detect liver and kidney function, urine protein, serum anti-nuclear antibodies, anti-dsDNA antibodies, anti-sm/nRNP antibodies, and observe the histopathological changes of the kidney and skin and the deposition of immune complexes. [Characteristics of the model] Compared with the control group, the anti-sm/nRNP antibody and anti-dsDNA antibody gradually increased in the model group at the 4th and 8th week. Serum creatinine began to appear abnormal at 6 weeks; serum albumin did not begin to decrease until 16 weeks, and was significantly lower than the control group at 18 weeks. After 6 weeks, the 24-hour urine protein content of rabbits was higher than that of the control group. After 8 weeks, there was a significant difference between the two groups, which gradually increased over time and was more than 5 times that of the control group. The degree of kidney disease in the model group was significantly higher than that in the control group. Membranous proliferative nephritis mainly involved glomeruli, renal tubules and renal interstitium. H&E staining showed that the renal lesions in the model group showed a diffuse increase, mainly glomerular and focal mesangial hyperplasia. Masson staining showed that collagen was deposited in the glomerulus, and immunohistochemistry showed that IgG was deposited in the mesangial region. Rabbits in the model group showed obvious skin damage, manifested as erythema, scaling and hair loss on the nape, back, toes, ears, etc. The rash gradually increased to the back and abdomen, and later became erythema infiltration and toe ulcers. H&E staining of skin pathological sections showed that the rabbits in the model group had significantly reduced hair follicles, liquefaction and degeneration of epidermal basal cells, and dilation and hyperemia of epidermal capillaries.

　　[Model Evaluation and Application] This model is used to study the causes of SLE and autoimmune nephritis.

　　Pristane, modeling of allogeneic lymphocytes or allogeneic hepatocyte plasma proteins is based on the same principle. In other words, it stimulates animals to cause abnormal and excessive immune responses, and its main feature is the appearance of various high-titer autoantibodies. It is related to immune complex, immune complex glomerulonephritis, arthritis, skin damage. The derived SLE model is easy to construct and low cost. Most SLE animal models use mice, but if the required sample size does not meet your research needs, rabbits can be used as SLE animal models. It should be noted that rabbits belong to inbred strains and individual differences are large, which should be considered in the experimental design.