| Objective:To establish a stable model of allograft transplantation in situ rabbit ears.To determine the relationship between the degree of rejection and expression of these factors by detecting a variety of cytokines expression of the rabbit ear transplantation group and control group (using immunosuppressant FK506) with acute rejection.Methods:1.Choose similar weight, healthy Livid blue rabbits and Japen big ear rabbits each 8 as experimental animals for allogeneic transplantation in situ rabbit ears. Referring to the method of establishment of rabbit ear transplantation model, to establish a stable model of allograft transplantation in situ rabbit ears.2.Will only 8 the rabbits after the operation were randomly divided into two groups: experimental group and control group (using immunosuppressant FK506), each group is divided into 6 h,12 h,24 h and 48 h four time points.3.To take four animals samples from each experimental group and control group after operation in 6h,12h,24h and 48h at four time points, the following materials and testing.① Using ELISA technology, to detect the expression changes of rabbit serum IL-2, IL-6 and IL-10;② Using flow cytometry to detect the content of CD4+ and CD8+T cell subsets in peripheral blood and to determine the changes of immune function after transplantation in rabbit ears;③ Using formalin to fix the rabbit ear tissue and detect it with HE staining. Observe morphological changes.Results:1. Transplantation group and FK506 group preoperative IL-2 (0.198 VS 0.327, P< 0.05), IL-6 (0.238 VS 0.235, P< 0.05), IL-10 (58.57 VS 59.28, P< 0.05), CD4+ (25.759 VS 16.641, P< 0.05) differences are significant, but there was no significant difference of CD8+level meaning (6.382 VS 5.481, P> 0.05).2. Transplantation group and FK506 group 2 (IL-6 h after surgery vs0.392 0.227, P < 0.05), IL-6 (0.324 vs 0.312, P< 0.05), IL-10 (66.88 vs 62.00, P< 0.05), CD4+ (28.612 vs 18.332, P< 0.05) differences are significant, but there was no significant difference of CD8+level meaning (7.513 vs 6.389, P>0.05).3. The transplant group and FK506 group 2 (IL-12 h after surgery vs0.453 0.404, P < 0.05), IL-6 (0.415 vs 0.408, P< 0.05), IL-10 (61.832 vs 68.375, P< 0.05), CD4+ (40.722 vs 20.475, P< 0.05) differences are significant, but there was no significant difference of CD8+level meaning (9.766 vs 9.058, P> 0.05).4. Transplantation group and FK506 group 24 h after surgery IL-2 (vs0.572 0.493, P < 0.05), IL-6 (0.483 vs 0.459, P< 0.05), IL-10 (58.24 vs 75.74, P< 0.05), CD4+ (60.549 vs 30.893, P< 0.05) differences are significant, but there was no significant difference of CD8+level meaning (10.387 vsl 1.032, P> 0.05).5. Transplantation group and FK506 group 48 h after surgery IL-2 (vs0.634 0.588, P < 0.05), IL-6 (0.541 vs 0.507, P< 0.05), IL-10 (60.799 vs 69.333, P< 0.05), CD4+ (90.478 vs 50.564, P< 0.05) differences are significant, but there was no significant difference of CD8+level meaning (12.139 vs 12.934, P> 0.05).Conclusion:1. Three cuff allograft transplantation model rabbit ears than traditional modeling is a kind of animal model of high repeatability, simple, economy, after the success of the allogeneic transplant rabbit ears, in a very short period of time can produce human acute rejection similar.2. The rabbit ears at different time points after transplantation IL-2, IL-6, IL-10 have obvious rise, FK506 have very good inhibitory effect, FK506 also inhibit rabbit ear the appreciation of CD4+T cells after transplantation, but has certain inhibitory effect on CD8+T cells, but not CD4+T change significantly, may it with CD4+T cells in the process of cellular immunity was first activated.3. The organization morphology observation confirmed FK506 on rabbit ear relieve acute rejection. |
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