动物造模,狼疮小鼠模型 z-bio 2021-10-19 1006

　　(1) Reproductive method (female DBA/2 x male C57BL/6J) F1 generation mice weighing about 16 g and 6 to 8 weeks old are injected into the DBA/2 lymphocytes of female mice and passed through the tail vein Injection guide model. At 12 weeks, first, DBA/2 mice were anesthetized, and the spleen, thymus and lymph nodes were aseptically separated at a ratio of 3:2:1. The tissue was ground in saline and passed through 150 μm and 70 μm nylon meshes. Observe the viability of the cells under a microscope and count the number of cells. The prepared live lymphocytes were injected into the body through the tail vein of F1 generation mice. The injection volume of each mouse is 50 x 1,000,000 live lymphocytes. The injection time was 0, 3, 7 and 10 days, and the observation time was 12 weeks. Urine of model mice was collected at a predetermined time of the experiment to measure urine protein concentration, and blood was used to measure serum creatinine (SCr), urea nitrogen (BUN), cholesterol (TC) and triglycerides (TG). in. ), albumin (ALB) and other indicators and indirect immunofluorescence to detect autoantibody dsDNA and observe under a fluorescence microscope. Take the kidneys of model mice, prepare frozen sections, conventional paraffin sections and ultrathin sections with an electron microscope, and then observe them with an optical microscope and a transmission electron microscope.

　　(2) Model characteristics The activity of the model mice began to decline from the 4th week after the injection of live lymphocytes, and the weight began to increase from the 8th week, and the hair was slightly black. Mice have subcutaneous edema, and some animals have ascites. Some mice will twitch. Ten weeks after the injection, the abdomen of the model mice was obviously enlarged, moving slowly and sluggish. Make sure that there is a large amount of clear ascites on the abdomen with naked eyes, the spleen and kidneys are obviously enlarged, and the kidneys are thinning. The separated serum is Cairo-like, the mouse model rate is 96%, and all unmodeled mice are male. The F1 generation mice developed autoantibodies and proteinuria 2 weeks after the injection of mother mouse lymphocytes. The F1 generation mice showed a slight increase in blood lipids, serum creatinine and urea nitrogen at 4 weeks under the microscope. The following is only the kidney The internal mesangial tissue is visible. The cells proliferated slightly without interstitial damage; at the 8th week, the blood biochemical indicators of the mice changed significantly, albumin and total protein increased, and after 12 weeks, albumin and total protein decreased. Observe under the microscope the proliferation of glomerular mesangial cells, inflammatory cell matrix infiltration and large amount of proteinuria into the renal tubules; at the 12th week, the mesangial glomerulus showed moderate to severe proliferation and glomerular disorder, similar to humans Type VI lupus nephritis, in which a large number of immune complexes are found under the endothelium, and the local or diffuse kidney is very small. In the course of 10-12 weeks, various observation indicators have changed significantly. Immunofluorescence showed that IgG, IgM and C3 were deposited along the capillary wall and mesangial area. The lupus mouse model established by this method is easy to create, develops rapidly, has typical pathological characteristics, and has a high model rate.

　　(3) Comparative Medicine Systemic lupus erythematosus (SLE) is an autoimmune disease that seriously endangers human health. The cause has not yet been elucidated. Choose the right ideal animal to further explore its etiology and preventive measures. This is very important. Mouse models of spontaneous lupus (such as NZB/NZW, MRL and BXSB) have been widely used in previous research reports. Although these mice can develop and develop SLE independently, they have the disadvantage of being time-consuming and difficult to control. The value of using experimental condition models that affect these conditions. Chronic graft-versus-host disease (CgVHD) lupus-like mouse model is an internationally recognized lupus mouse model, which can be induced by experimental methods, the experimental conditions are easy to control, the onset is early, 12 weeks after induction, the typical lupus nephritis may appear The pathological changes, which are similar to the typical symptoms of human lupus nephritis and lupus nephritis, are particularly suitable for research. The etiology of the mouse model of chronic graft versus host disease (cGVHD) is characterized by lymphoplasty, which may produce autoantibodies similar to clinical SLE patients, and severe immune complex-mediated kidney disease. The etiology of this model was induced by transplanting parental lymphocytes into F1 generation mice, and its etiology was partial incompatibility of MHC. The genotype of the female DBA/2 in this model is H-2d, the genotype of the parent mouse (such as C57BL/6 or C57BL/10) is H-2b, and the genotype of the F1 generation mouse obtained after mating is H- 2b. H.-2d/b, the part of the MHC class is the same. Donor DBA/2 mice lack the same cytotoxic CD8 + T cells as anti-F1 generation mouse T cells, so MHC-type CD8 + T cells are inactive, have INF-γ content in the body, and are accompanied by CTL And CTL reduction. Cells; in vivo anti-allogeneic MHC-IICD4 + T cells activate receptor B cells, thereby causing B cell proliferation in F1 generation mice, thereby producing autoantibodies, and ultimately leading to SLE-like expression. The lupus-like mouse model of chronic graft-versus-host disease established by this method is similar to clinical human lupus nephritis and is an ideal lupus nephritis model.