| 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.â‘ Using ELISA technology, to detect the expression changes of rabbit serum IL-2, IL-6 and IL-10;â‘¡ 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;â‘¢ 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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