Immune Effection Of Artesunate During Acute Rejection In Small Bowel Transplantation Of Rats

Objectives1. To learn the operation of model of rats SBT and established the model skillful.2. To investigate the expression of blood serum IL-2, IFN-γand its diagnosis value in acute rejection in small bowel transplantation of rats.3. To investigate the role of Artesunate in acute rejection of rat small bowel transplantation and it's mechanism of immune suppression.Methods1. Animal choice. Inbred male Wistar rats and F344/N rats with body mass of 250±30g were random divided into control group (n=6), isograft group (n=18), allograft group (n=18) and receive treatment allograft group (n=18.This group was administrated Artesunate by intraperitoneal injection at a dose 60mg/kg/day from 7 days before operation to 7 days after operation).2. Animal model.The model was established with the method of Li Jie-shou. Use the 1% ketamine with 10mg/100g body mass intraperitoneal injection to anesthetize. The small bowel of donor was gotten the 7~10 cm jejunum from the 1cm below the Treitz. The superior mesenteric artery of grafts was anastomosed with the abdominal aorta of the recipients. The portal vein of grafts was anastomosed with the left renal veins of recipients with method of duct of Cuff. The grafts were harvested on the postoperative day (POD) 3, 5, 7 and were observed by pathohistology methods, and the expression of IL-2 and IFN-γof blood serum were detected by enzyme-labeled immunosorbent assay (ELISA) kits.3. Histopathology assessments. Tissue samples were fixed in 4% buffered formalin and prepared for hematoxylin–eosin (H&E) staining. Morphologic changes in the graft jejunums were evaluated with light microscopy to definite the happen of graft rejective reaction and its level.4. ELISA Analysis. IL-2 and IFN-γof blood serum in POD3, 5, 7 were detected by enzyme-labeled immunosorbent assay (ELISA) kits of Wuhan Boster Corporation.Results1. SBT model of rats were successfully established. The model showed that mild acute rejetion occurred on POD 3, moderate acute rejection on POD 5, severe acute rejection on POD 7 in allograft group. The control group and isograft group has no evidence of acute rejection. Some of the treatment group showed mild rejection on POD 7.2. path-histology examination. The acute rejection occurred on POD3 in allograft group and gradually aggravated on POD5, 7 while the control group and isograft group have no evidence of acute rejection. Treatment group showed mild rejection on POD 7 although it's not severe.3. IL-2. The level of blood serum IL-2 were significantly higher on POD 3, 7 and reached the peak on POD 5 in allograft group than the other three groups(P<0.01). The expression of blood serum IL-2 in treatment group were higher than isograft group too, but with no significant difference (P>0.05).4. IFN-γ.The level of blood serum IFN-γwere gradually raise up on POD 3, 5 and reached the highest on POD 7 and higher than the other three groups(P<0.05). Control group and isograft group have no significance (P>0.05). The level of IFN-γof treatment group were higher than control group and isograft groupuring (P>0.05).Conclusions1. IL-2 and IFN-γplay an important role in small intestinal allograft acute rejection. To detect the expression level of blood serum IL-2,IFN-γmay be a important indicator for morning diagnosis of acute rejection of rat small intestinal transplantation.2. The Artesunate can suppress the acute rejection of small intestinal allograft. It's mechanism of immune suppression may be through suppress the expression and secretion of cytokine such as IL-2,IFN-γ.