| Objective:To establish a stable rat model of acute hepatic failure and a rat model of heterotopic auxiliary liver transplantation in spleen fossa with splenectomy. To observate the pathological changes of graft liver after operation. useing semi-quantitative RT-PCR to monitor the expression of Perforin mRNA in peripheral blood. To explore the relationship between the expression of perforin mRNA and acute hepatic allograft rejection. To evaluate the therapy of making the early diagnosis of acute rejection.Methods:This design of experiment use adult healthy clean rats, whose weights ranged from 250g to 320g. They was divide into three groups:Group A:blank group (only undergoing left nephrectomy and lienectomy with SD rats); Group B:allograft transplantation group (graft liver came form Wistar rats, host liver came form SD rats) and treated with cyclosporine; Group C, allograft transplantation group and untreated controls. To select rats according to above groups, recipient rats underwent 85% partial (right up lobe,middle lobe,left lateral lobe,left medial lobe) hepatectomy, the graft liver which was reduced to about 30% of the original size, was inplanted into the spleen fossa of the recipient's abdomen. The samples were obtained at different times, which were timed on the first day, the third day, the fifth day, the seventh day and the tenth day postoperative. To select six rats at every observation point semi-quantitative was used to measure the expression of perforin mRNA in peripheral blood, and the pathological changes of the graft were observed as well.Results:1. To establish a rat model of heterotopic auxiliary liver transplantation in spleen fossa with splenectomy:The operations were performed at 110±10min, To control the warn ischemia time within 5min and the cold ischemia time within 50min. the total sueeessful rate was above 88.3%. The common cause of death after operation are as flollow:anastomotic haemorrhage,leakage of the bile-duct anastomosis,portal vein phlebothrombosis,bowel obstruction and so on.2. Postoperative acute rejection could be reflected through the pathological changes of graft liver. During acute rejection in rat auxiliary liver transplantation, The pathological changes of graft liver was similar to the expression of Perform mRNA in peripheral blood. Perform mRNA expression of all specimens could not be detected at 1 day after transplantation. Group A had not monocyte and lymphocyte could be seen in the portal tracts and there were not denaturation of hepatocyte, Perforin mRNA in peripheral blood could not be detected at each different time points; Group B, from 3 to 10 day, CsA abated cellular infiltration, but hepatocyte necrosisand hemorrhage remained unchanged, Perform mRNA continuously expressed in a low level; Group C, the pathological results showed that except of the first day, mild-moderate and severe acute rejection occurred on the third,fifth, and seventh to tenth day after operation respectively, Perform mRNA expression began to up-regulate and increasingly elevated after 3 days later. Levels of expression was significantly increased after 5 days later and remained in high level during 7 to 10 days. The result show it found the expression of Perforin mRNA in peripheral blood both Group B and gooup C, but Group A. Compared the Group B with the group C, there were significantly statistical differences between different time points (P<0.05).Conclusions:1. We can established an scientific and advisable animal model that heterotopic auxiliary liver transplantation in spleen fossa with splenectomy and with portal vein arterialization in rat. The graft liver play an exact support role for the rat on passing the acute hepatic failure time.2. According to monitor the changes of the expression of Perform mRNA in peripheral blood in different time point, we find the change was paralleled between the pathological changes of graft liver and it. While acute rejection in rat liver transplantation was occurring, the expression levels of perform mRNA between grafts and Peripheral blood, which was coherent with the level of the severity degree of histologic damage. So through RT-PCR to measure the expression of perform mRNA in the peripheral blood, to offer the scientific conformity system that can be operated easily to diagnose acute rejection of renal allograft, and it is a sensitive indicator of diagnosing acute rejection.3. Cyclosporine A can decrease the number of infiltrating cells and degrade the expression levels of perform mRNA. It has palpable effect to reduce acute rejection.4. We need a further research on the problems such as the cooperation and competition relationship between the graft liver and the host liver and the affection on the livers after a long application of immunosuppressant.
|
PDF Full Text Request