Influences Of Exogenous IL-10 And TGF-β1 To The Acute Rejection Of Small Intestinal Allograft In Rats

of exogenous TGF- P 1 on the acute rejection of small intestinal allograft.Methods Establish the model of intestinal allograft of rat. Rats are divided into three groups. Group I: Wistar rats —? SD rats Small intestinal transplantation. Group II: Wistar rats —> SD rats Small intestinal transplantation + CsA. Group III: Wistar rats —? SD rats Small intestinal transplantation + rhTGF-P 1. Each group has six rats. We get part of allograft at 3rd day, 5th day, and 7th day after operation. We investigated the pathologic changes, and applied immunohistochemical method to detect the expressions of CD4 and CD8. The transcriptions of IL-2, IFN-y mRNA were detected by Semi-quantity RT-PCR.Results1 Pathologic changes: There were acute rejections after operation in group I, and became severer as time passed. It had grade 0- I acute rejections at 3rd day, grade II acute rejections at 5th day and grade III acute rejections at 7th day. There nearly haven't acute rejections in group II. Only grade 0- I rejections happened at 7th day. There were slight rejections after operation in group III, it had no rejection at 3rd. But it had grade I rejections at 5l and grade II rejection at 7th.2 Results of Immunohistochemical detection: CD4 and CD8 mostly expressed at the basal layer of small intestine. They expressed only a little in normal small intestine. CD4 and CD8 all increased after operation. Group I increased higher than group II and group III. Their differences had statistic significance (p<0.05)3 Results of RT-PCR: The expressions of IL-2 mRNA and IFN-y mRNA increased after operation in group I at 3rd day after operation, and reached the peak at 5th, 7th day. The expressions of IL-2 mRNA and IFN-y mRNA changed a little in group II, compared with group I ,they were lower, and their differences had statistic significance(/?<0.05). The expressions of IL-2 mRNA and IFN-y mRNA increased after operation inischemia time as possible. The donor small bowel is harvested based on a vascular pedicle consisting of the mesenteric artery with a part of aorta, and the portal vein that is transected above its bifurcation. Get the graft and make it suitable for transplantation. Recipient operation: After the inferior vena cava and the aorta are released from surrounding connective tissue and cross-clamped with microvessel clips below the left renal vessels, the donor portal vein is placed end to side into the recipient inferior vena cava first. Secondly, the arterial end-to-side anastomosis is performed. Both the ends of graft are exteriorized and sutured to the abdominal wall of recipient as stomas. The abdominal wall is then closed. Anastomose of blood vessel is under the surgical microscope which can magnify 10 times.Results This experiment is divided in to two phases, totally performed 75 operations. The first phase is to acquaint with anatomical structure and skill of operation. It is not calculated. The second phase is formal experiment, totally performed 35 operations. Succeeded 29 cases, defeated 6 cases. The rate of success is 82.8%. It is similar to other reports.The operation time of donor is about 70min±10min. The operation time of receptor is about 80min±10min, in which the time of blockade of blood vessel is 45-50min. The anastomose time of artery is about 20min.The anastomose time of vein is about 20min. The time of hot ischemia is 1min. cold ischemia time is 30min, and warm ischemia time is 45min.Conclusions1 The model of segmental small intestinal transplantation in rat is a nicer model to research the rejection of transplanted small intestine. Its rate of success is high.2 Surgical technic is a key point of small intestinal transplantation inrat. It needs experience, and skill.3 we must prevent lose too much blood and body fluid in the operation.4 When get intestine from donor, we must prevent injure the small intestine, and shorten the time of hot ischemia.PART TWOINFLUENCE OF EXOGENOUS IL-10 TO THE ACUTE REJECTION OF SMALL INTESTINAL ALLOGRAFT IN RATSObjective Organ transplantation is radical therapy for severe diseases of many organs. As a high immunogenic organ, intestine has lots of lymphatic tissue. Cases of intestinal transplantation are fewer than other organ transplantation. Its curative effect is not satisfied. And the rate of survival is lower. The major reason of frustration of intestinal transplantation is intensive rejection. IL-2 and IFN-y are cytokine of Thl. IL-2 is an important mediator to speed proliferation and differentiation of T cell. It stimulates MHC- II to express on T cell, and stimulates T cell to produce lots of lymphokine. It can stimulate NK cell to proliferate and activate macrophage. IFN-y is a potent activator of macrophage. It can promote differentiation of T cell and B cell. It promotes CTL to mature and stimulates B cell to secrete antibody. Both of them express obviously in acute rejection. IL-10 is a cytokine of TH2. It can restrain the activation of some immunocyte and the production of cytokine. It has closely relationship with toleration of graft. This experiment is to observe the influence of exogenous IL-10 on the acute rejection of small intestinal allograft in rats, and to investigate the relationship between exogenous IL-10 and immunologic tolerance of specific antigen.Methods Establish the model of small intestinal allograft in rats. Rats are divided into three groups. Group I: Wistar rats —*■ SD rats Small intestinal transplantation. Group II: Wistar rats —>■ SD rats Small intestinal transplantation + CsA. Group III: Wistar rats —? SD rats Small intestinal transplantation + rrIL-10. Each group has six rats. We get part of allograft at 3rd day, 5th day, and 7th day after operation. We investigated the pathologic changes, and applied immunohistochemical method to detect the expressions of CD4 and CD8. The transcriptions of IL-2, IFN-y mRNA were detected by Semi-quantity RT-PCR.Results1 Pathologic change: There were grade 0- I acute rejections at 3rd day, grade II at 5th day and became III at 7th day at group I. There nearly haven't acute rejection in group II. Only grade 0- I rejections at 7th day. There were slight rejections after operation in group III, no rejection at 3rd, grade I rejection at 5th, grade II rejection at 7th.2 Results of Immunohistochemical detection: CD4 and CD8 mostly expressed at the basal layer of small intestine. They expressed only a little in normal small intestine. CD4 and CD8 all increased after operation. Group I increased higher than groupII and group III. Their differences had statistic significance (p<0.05)3 Results of RT-PCR: The expressions of IL-2 mRNA and IFN-y mRNA increased after operation in group I at 3rd day after operation, and reached the peak at 5th, 7th day. The expressions of IL-2 mRNA and IFN-y mRNA changed a little in group II, compared with group I , they were lower, and their differences had statistic significance O<0.05). The expressions of IL-2 mRNA and IFN-y mRNA increased after operation in group III at 3rd day after operation, and reached the peak at 5th,7th day. but they are lower compared with group I . Their differences had statistic significance (p<0.05). The expressions of IL-2 and IFN-y mRNAchanged earlier than pathologic changes. Conclusions1 Typical acute rejection can be observed in the model of Small intestinal transplantations from Wistar rats to SD rats, and its occurrence is stable.2 Expressions of CD4, CD8 in the allograft are correlated with acute rejections of intestinal allograft in rat.3 The expressions of IL-2> IFN-y mRNA can represent the degree of acute rejections of intestinal allograft.4 Exogenous IL-10 can restrain the acute rejections of small intestinal allograft in rats. PART THREEINFLUENCE OF EXOGENOUS TGF- & 1 TO THE ACUTE REJECTION OF SMALL INTESTINAL ALLOGRAFT IN RATSObjective Rejection of transplantation is a major obstacle to influence the survival of graft. Effectively restrain the rejection of graft is a radical problem to improve the success rate and prolong the surviving time of graft. TGF- 3 extensively participates in modulation of cytokine in immune system, and influence the proliferation and differentiation of lymphocyte. It interferes with the production and change of immunoglobulin. TGF- P influences IL-1, IL-2, IFN- V by direct or indirect ways. It plays a negative modulation on the balance of immunity. Lately TGF- P is used as a powerful immunosuppressive factor to restrain immunologic rejection and get a satisfied result. Its concrete mechanism has become a hotspot to study. This experiment is to observe the influenceof exogenous TGF- P 1 on the acute rejection of small intestinal allograft.Methods Establish the model of intestinal allograft of rat. Rats are divided into three groups. Group I: Wistar rats —? SD rats Small intestinal transplantation. Group II: Wistar rats —> SD rats Small intestinal transplantation + CsA. Group III: Wistar rats —? SD rats Small intestinal transplantation + rhTGF-P 1. Each group has six rats. We get part of allograft at 3rd day, 5th day, and 7th day after operation. We investigated the pathologic changes, and applied immunohistochemical method to detect the expressions of CD4 and CD8. The transcriptions of IL-2, IFN-y mRNA were detected by Semi-quantity RT-PCR.Results1 Pathologic changes: There were acute rejections after operation in group I, and became severer as time passed. It had grade 0- I acute rejections at 3rd day, grade II acute rejections at 5th day and grade III acute rejections at 7* day. There nearly haven't acute rejections in group II. Only grade 0- I rejections happened at 7th day. There were slight rejections after operation in group III, it had no rejection at 3rd. But it had grade I rejections at 5l and grade II rejection at 7th.2 Results of Immunohistochemical detection: CD4 and CD8 mostly expressed at the basal layer of small intestine. They expressed only a little in normal small intestine. CD4 and CD8 all increased after operation. Group I increased higher than group II and group III. Their differences had statistic significance (p<0.05)3 Results of RT-PCR: The expressions of IL-2 mRNA and IFN-y mRNA increased after operation in group I at 3rd day after operation, and reached the peak at 5th, 7th day. The expressions of IL-2 mRNA and IFN-y mRNA changed a little in group II, compared with group I ,they were lower, and their differences had statistic significance(/?<0.05). The expressions of IL-2 mRNA and IFN-y mRNA increased after operation in...