TGF-β1修饰MSC诱导移植胰腺免疫耐受的实验研究

发布时间：2018-07-10 02:16

本文选题：转化生长因子-β1 + 骨髓间充质干细胞　；参考：《昆明医科大学》2015年硕士论文

【摘要】：目的通过注射外源性转化生长因子β1(TGF-β1)转染骨髓间充质干细胞(MSC)予兔胰腺移植(PTA)模型受体,探讨TGF-β1修饰MSC能否诱导移植胰腺免疫耐受。材料和方法1.采用健康幼龄日本大耳白兔24只,随机抽出12只作单独胰腺移植(PTA)的供体,余12只制作糖尿病模型,模型成功后成为PTA的受体。将受体分成(A、B、C、D)组,给予不同处理,A组：受体胰腺移植前5天给予MSC静脉输注,B组：受体胰腺移植前5天给予TGF-β1转染MSC静脉输注,C组：受体胰腺移植前5天只给TGF-β1,D组：空白组,受体胰移植前不行干预。2.对供体进行分离、灌注、切取、修剪供体胰腺；分离受体一侧肾后,阻断肾动静脉,用袖套法完成供体门静脉与受体肾静脉的吻合,用缝合法完成腹腔干动脉与肾动脉吻合,输尿管和胰管置管行膀胱内引流。3.对术后受体进行处理,取各组移植兔术后不同时间的静脉血及尿液来测定内外分泌功能,并对术后死亡兔子进行病理检查分析。4.对术后受体进行处理,取各组移植兔术后第1、3天共2个时间段静脉血,应用流式细胞仪检测CD4+T细胞和CD8+T细胞百分率,并计算其比值及CD4+CD25+Treg细胞的比例。结果1. 12只供体顺利完成胰腺切取术,12只受体成功制作糖尿病模型并顺利完成胰腺移植术；供体胰腺移植后迅速恢复血液循环,无出血及漏血；术后1例死于静脉血栓形成,实验组的9只受体术后平均生存期(8.0±0.7)d,对照组的3只受体术后平均生存期(7.0±0.2)d,成功建立兔胰腺移植模型。2.术后实验组胰腺内外分泌功能在存活期间比对照组好,差异有统计学意义(P0.05),病理检查结果显示明显比对照组排斥反应低。3.术后MSC组(n=3)比未干预组(n=3) CD4+T细胞数和CD4+CD25+Treg细胞数比例高,差异有统计学意义(P0.05)。4.术后TGF-β1转染MSC组(n=3)比未干预组(n=3) CD4+T细胞数和CD4+CD25+Treg细胞数比例低,差异有统计学意义(P0.05)。5.术后TGF-β1组(n=3)比未干预组(n=3) CD4+T细胞数和CD4+CD25+Treg细胞数比例高,差异有统计学意义(P0.05)。6.术后TGF-β1转染MSC组(n=3)比MSC组(n=3)、TGF-β1组(n=3) CD4+T细胞数和CD4+CD25+Treg细胞数比例高,差异有统计学意义(P0.05)。结论TGF-β1转染MSC能够调节动物胰腺移植免疫反应,可减轻排斥反应,较单独使用MSC或TGF-β1免疫效果明显。
[Abstract]:Objective to investigate whether TGF- 尾 1 modified mesenchymal stem cells (MSC) can induce pancreatic allograft tolerance by injecting exogenous transforming growth factor 尾 1 (TGF- 尾 1) into rabbit pancreatic transplantation (PTA) model receptor. Materials and methods 1. A total of 24 healthy young Japanese large ear white rabbits were randomly selected as donors for pancreas transplantation (PTA) and 12 diabetic models were established. The recipients were divided into two groups: group A: group B received MSC intravenous infusion 5 days before pancreas transplantation: group C received TGF- 尾 1 transfection into MSC 5 days prior to pancreatic transplantation: group D: group B: TGF- 尾 1C: group D: group B: TGF- 尾 1D: group C: TGF- 尾 1D: group D: group B: TGF- 尾 1D: group C: TGF- 尾 1D; The receptor did not intervene before pancreas transplantation. The donor was separated, perfused, cut off and pruned the donor pancreas, after separating the recipient kidney, the renal artery and vein were blocked, the donor portal vein and the recipient renal vein were anastomosed with cuff method, and the abdominal trunk artery and renal artery were anastomosed by suture. Ureteral and pancreatic duct catheters were placed for intravesical drainage. The recipient was treated and the venous blood and urine were taken from each group of rabbits at different time after transplantation to determine the function of exocrine and exocrine secretion, and the pathological examination and analysis of the dead rabbits after operation were made. 4. The recipients were treated and the venous blood was taken from the rabbits at the 1st day after transplantation. The percentages of CD4 T cells and CD8 T cells were measured by flow cytometry, and the ratio of CD4 CD25 Treg cells were calculated. Result 1. 12 donors successfully completed pancreatic excision and 12 recipients successfully made diabetic models and successfully completed pancreas transplantation; donor pancreas transplantation quickly recovered blood circulation without bleeding or bleeding; 1 case died of venous thrombosis after operation. The mean survival time of 9 recipients was (8.0 卤0.7) days in the experimental group and (7.0 卤0.2) days in the control group. The exocrine and exocrine pancreatic function in the experimental group was better than that in the control group during the survival period, the difference was statistically significant (P0.05). The pathological results showed that the rejection was significantly lower than that in the control group. The percentage of CD4 T cells and CD4 CD25 Treg cells in MSC group was higher than that in non-intervention group (P0.05). The percentage of CD4 T cells and CD4 CD25 Treg cells in MSC transfected with TGF- 尾 1 was significantly lower than that in control group (P0.05). The ratio of CD4 T cells and CD4 CD25 Treg cells in TGF- 尾 1 group was higher than that in non-intervention group (P0.05). The percentage of CD4 T cells and CD4 CD25 Treg cells in TGF- 尾 1 transfected MSC group was higher than that in MSC group after operation (P0.05). Conclusion transfection of MSC with TGF- 尾 1 can modulate the immune response of pancreas transplantation and alleviate rejection, which is more effective than that of MSC or TGF- 尾 1 alone.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R657.5

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