重组hIL-10抗家兔皮肤移植排斥反应及其T淋巴细胞亚群的变化

发布时间：2018-05-26 13:54

本文选题：重组人白介素-10 + 家兔　；参考：《遵义医学院》2010年硕士论文

【摘要】： 目的：构建同种异体家兔皮肤移植的动物模型；经重组hIL-10蛋白作用后,观察家兔皮肤移植后一般情况、皮片出现排斥的时间和皮片存活的时间；观察家兔皮肤移植后T淋巴细胞亚群的变化,以探讨重组hIL-10在抗皮肤移植机制中的作用。方法：1.家兔皮肤移植实验：受体家兔随机分组,分6组,每组3只,共18只；供体家兔共18只,随机抽取与受体配对。移植前先背部剃毛,切除1.5cm×1.5cm大小面积的中厚皮片进行移植,每只家兔移植2片皮片,缝合固定及单笼饲养。移植前1d开始肌肉注射药物,连续注射,观察术后各组家兔一般情况及皮片的情况。2.流式细胞检测：用荧光双标、直接法分别检测移植前1d,移植后1d,4d,7d,14d,21d,28d各时间段各组家兔外周血中CD3+、CD3+CD4+、CD3+CD8+T淋巴细胞数的百分比以及CD3+CD4+/CD3+CD8+比值。结果1.家兔情况：大剂量CsA组术后出现饮食、精神状态欠佳、毛发脱落等现象,其余组均未出现上述现象；皮片情况：术后3天去加压包扎,见敷料干燥,皮片柔软红润,张力适中,周边整齐无渗出,术后4天开始阴性对照组出现排斥现象。2.皮肤开始出现排斥反应时间：低剂量hIL-10组为6.67±2.08 d,高剂量rhIL-10组为9±3.60d,低剂量CsA组为12±1.0 d,高剂量CsA组为19.67±1.15 d,联合组为16.67±1.15 d,阴性对照组为4.33±0.58 d,除低剂量rhIL-10组外,各组和阴性对照组之间差异都有显著意义(p0.05)。3.皮肤存活时间：低剂量hIL-10组为13.33±3.05 d,高剂量rhIL-10组为16±2.0 d,低剂量CsA组为15.67±0.57 d,高剂量CsA组为21.33±2.52 d,联合组为19±1.73 d,阴性对照组7.67±1.15 d,各实验组与阴性对照组之间差异均有显著意义(p0.05)。4.在发生急性排斥反应时,CD3+CD4+T淋巴细胞数和CD3+CD4+/CD3+CD8+比值均升高,而CD3+CD8+T淋巴数变化与排斥反应的发生无明显关系。结论：1.动物实验证实rhIL-10对同种异体家兔皮肤移植排斥有抑制作用,可推迟皮片排斥反应出现的时间,可延长移植皮片存活时间,并与CsA有协同作用。2.rhIL-10抑制CD3+CD4+T淋巴细胞活化,通过抑制细胞活化,降低IL-2,IFN-γ的产生,诱导Th1向Th2转化,延缓排斥反应的发生。3.CD3+T淋巴数在移植术后均有不同程度升高,说明在受到外原刺激后引起CD3+活化、增生,CD3+CD4+/CD3+CD8+比值与排斥反应开始出现时间及皮片存活时间均有一定的关系,比值在皮片开始出现排斥反应前升高,在皮片坏死时达到峰值,说明该比值可以监测排斥反应的发生。
[Abstract]:Objective: to establish an animal model of skin allograft in rabbits, observe the general situation after skin transplantation, the time of skin graft rejection and the survival time of skin graft after the action of recombinant hIL-10 protein. To observe the changes of T lymphocyte subsets after skin transplantation in rabbits and to explore the role of recombinant hIL-10 in the mechanism of anti skin transplantation. Method 1: 1. Rabbit skin transplantation experiment: the recipient rabbits were randomly divided into 6 groups with 3 rabbits in each group and 18 donor rabbits randomly selected and matched with the recipients. Before transplantation, the back was shaved and the 1.5cm 脳 1.5cm area of moderate thickness skin grafts were removed. Each rabbit was grafted with 2 skin grafts, fixed by suture and raised in a single cage. One day before transplantation, the drug was injected intramuscularly and continuously. The general situation and skin grafts of each group were observed. Flow cytometry: the percentage of CD3 CD4 CD3 CD8 T lymphocytes in peripheral blood and the ratio of CD3 CD4 / CD3 / CD3 CD8 in peripheral blood of rabbits in each group were detected by fluorescence double labeling and direct method 1 day before transplantation and 1 day after transplantation. Result 1. The condition of rabbits: diet, poor mental state, hair loss and other phenomena were not found in the high dose CsA group, but the skin grafts were not; the skin grafts were decompressed 3 days after operation, the dressing was dry, the skin was soft and ruddy, and the tension was moderate, the skin was soft and ruddy, and the tension was moderate. Peripheral neatly no exudation, 4 days after the negative control group appeared rejection phenomenon. 2. The reaction time of skin rejection was 6.67 卤2.08 days in low dose hIL-10 group, 9 卤3.60 days in high dose rhIL-10 group, 12 卤1.0 days in low dose CsA group, 19.67 卤1.15 days in high dose CsA group, 16.67 卤1.15 days in combination group and 4.33 卤0.58 days in negative control group. There was significant difference between each group and negative control group (P 0.05). The skin survival time was 13.33 卤3.05 days in the low dose hIL-10 group, 16 卤2.0 days in the high dose rhIL-10 group, 15.67 卤0.57 days in the low dose CsA group, 21.33 卤2.52 days in the high dose CsA group, 19 卤1.73 days in the combined group and 7.67 卤1.15 days in the negative control group. In acute rejection, the number of CD3 CD4 T lymphocytes and the ratio of CD3 CD4 / CD3 CD8 were increased, but the changes of CD3 CD8 T lymphatic number were not related to the occurrence of rejection. Conclusion 1. Animal experiments showed that rhIL-10 could inhibit skin allograft rejection in rabbits, delay the occurrence of skin graft rejection, prolong the survival time of skin graft, and inhibit the activation of CD3 CD4 T lymphocytes. 2. RhIL-10 could inhibit the activation of CD3 CD4 T lymphocytes. By inhibiting the activation of cells, decreasing the production of IL-2IFN- 纬, inducing the transformation of Th1 to Th2, delaying the occurrence of rejection, the number of CD3T lymphoid cells increased in varying degrees after transplantation, which indicated that the activation of CD3 was induced by exogenous stimulation. The ratio of CD3 CD4 / CD3 CD8 was correlated with the onset time of rejection and the survival time of skin graft. This ratio can be used to monitor the occurrence of rejection.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

【引证文献】