The Expression Of Th22, Th17 Cells And Related Cytokines In The Models Of The Orthotopic Liver Transplantation Of Rats With Acute Rejection

Section one Establishing and Evaluation of Acute rejection after orthotopic liver transplantation model of ratsObject:To establish the orthotopic liver transplantation (OLT) in the rats from which we could explore the mechanism and importance of graft rejection in liver transplantation. At the same time, we can establish the stable and effective animal models for subsequent tests.Methods:The male Sprague-Dawley (SD) rats and Wistar rats were selected as donors and receptors respectively. The rats applied for the production were divided randomly into three groups including control group, isograft group (Wistar-Wistar) and allograft group (SD-Wistar group). There were thirty-six cases of the orthotopic liver transplantation model of rats established with the technique of "two cuff method". We compare the survival rate and quality of rats in different groups, the pathological changes of liver with HE staining and the content of NO in serum with ELISA were studied furthermore. We assessed ultimately the effect of the liver transplantation model with acute rejection.Results:①The surgery success rate was 91.7%(33/36) under the condition of immunosuppressant. There was one case of isograft rats died in the operation due to bleeding and pneumothorax, two cases of allograft rats died because of bleeding of the inferior vena cava.Otherwise the rats of control group have no death. The 14 days postoperative survival rate of homologous transplantation group rats was 100%, and the median survival was 105 days. But the median survival was 6 days of different transplantation group.② The content of serum NO levels showed significantly different in three days of operation. The serum NO of allograft rats were (306.67±14.36)pg/ml and were significantly higher than the other two groups (P< 0.05). The serum NO of isograft rats was in the middle compare with the other two groups, and also was different from control group(60.77±13.66)pg/ml.③ There was no obvious inflammatory cell infiltration in the liver of control group and the structure of liver tissue was normal on the whole. It was 0 grade according to Williams standards. The rats of isograft group were 0 to one grade according to Williams standards because of showing a few inflammatory cells infiltrated in portal area.Yet allograft group appeared significant pathological changes such as large number of lymphocyte infiltration involving the bile duct, blood vessels, lobular integrity disappearing, accompanied by focal hemorrhage, necrosis, so it was 3 grade of Williams standard.Conclusions:The technology of "two cuff method" can simplify the operation procedure and improve success of surgery, and it maybe offer a stability of animal model for the next experiments. it can establish a stable model of acute graft rejection with the combination of Wistar-SD rats with immunosuppressant,.Section two The expression of Th22, Th17 cells and related cytokines in acute rejection after orthotopic liver transplantation of ratsObject:To observe of the changes of Th22 and Th17 and cytokines which related to in the rat liver with acute immune rejection after received the operation of orthotopic liver transplantation discussing the relation between the pathogenesis of acute rejection and liver transplantation furtherly.Methods:We established the model of orthotopic liver transplantation with The male Sprague-Dawley (SD) rats and Wistar rats that were acted as donors and receptors by the technology of "two cuff method". The amounts of Th22 and Th17 in the different groups were detected with flow cytometry, in the meantime,the mRNA and expression of protein were detected with reverse transcription-polymerase chain reaction and western blotting respectively.Resuts:The models of liver transplantation with acute immune rejection were successfully established. The numbers of Th22 and Th17 cells of rats in allograft group that were accepted the operation of orthotopic liver transplantation were (2.85±0.95) and (3.56±0.93), which were significantly higher than isograft group and contral group, (P <0.05).And the mRNA and expression of protein of IL-17and IL-22 were also significantly higher after seven days of postoperation compared with the other groups, (P < 0.05).Conclusions:The Th22 and Th17 cells and whose cytokines related with may play an important role during in the occurrence and development in rats with acute immune rejection received liver transplantation. The indexes above maybe prove to be the new indicators in clinical diagnosis of liver transplantation with acute rejection.