| [Objective]1. To establish a simple and feasible, stable, reproducible rat autologous orthotopic liver transplantation model, to provide animal models for the study after liver transplantation for acute kidney injury.2. Investigate the influence of perfusion in orthotopic liver transplantation for acute kidney injury in rats, and to provide experimental basis for application in clinical liver transplantation.[Methods]1. Autotransplantation in rats orthotopic liver transplantation model:Take24SD rats were randomly divided into model group18and the sham group6. Model group based on the forceps, the use of the perfusate through the portal vein infusion method for establishment of rat autologous orthotopic liver transplantation model. After the establishment of animal models of success1day,1week, blood samples were collected in January of liver function and liver tissue for pathological observation and weighing.2. Autologous orthotopic liver transplantation in different perfusion for acute kidney injury:Take54SD rats were randomly divided into three groups:the retrograde perfusion group24is perfusion group24with the sham group6. The first two groups on the basis of the establishment of self-donor liver transplantation model, the retrograde perfusion group were reverse perfusion through the inferior vena cava, first open the inferior vena cava, and then open the portal vein and hepatic artery; is reperfusion group using the conventional forward through the portal vein perfusion, which in turn open the portal vein, hepatic artery and inferior vena cava. The sham operation group only free portal hepatis the portal vein, hepatic artery and hepatic, inferior vena cava, not blocked. In group treatment,6h,12h,24h,48h animal blood line serum creatinine, blood urea nitrogen, cystatin C, kidney injury molecule-1determination, to take the kidney tissues pathology and kidney damage molecule-1expression in the observation of the situation.3. Statistical analysis:The application of statistical software SPSS13.0analysis of measurement data, mean±tandard deviation to describe multiple sets of measurement data were compared using single factor analysis of variance, further pairwise comparisons using the SNK test method; rate compared using the χ2test or Fisher exact test,P<0.05for the difference was statistically significant.[Results]1. Autotransplantation in rats orthotopic liver transplantation modelModel group the success rate of94%(17/18), sham operation group modeling success rate of100%(6/6). Perfusion effects in the model group, surgery, operation time was (10±3) min, bleeding was about0.2-0.5ml, postoperative anesthesia after20min can stand. Postoperative day1, model group, ALT and AST were higher than sham operation group (P<0.05); after1week in January, the serum levels of ALB,, the ALT and AST, body weight had no significant difference (P>0.05). Liver HE staining: after one day, the model group, moderate edema of the liver cells, fatty degeneration, sinusoidal expansion significantly, the apparent expansion of the central venous congestion, portal area interlobular veins were expanded, more inflammatory cell infiltration. After one week in January, the model group, no obvious abnormalities. Sham operation group showed no obvious abnormalities.2. Autologous orthotopic liver transplantation in different perfusion for acute kidney injury2.1Content in serum creatinine (Scr), blood urea nitrogen (BUN), cystatin C (cystatin C) and renal injury molecule-1(KIM-1):Serum creatinine (Scr), blood urea nitrogen (BUN), cystatin C (cystatin C) and renal injury molecule-1(KIM-1) content. Compared with the sham group, the retrograde perfusion group perfusion group in the postoperative6h,12h,24h values of Scr and BUN values were higher, the difference was statistically significant (P<0.05). Retrograde perfusion group after6h,12h,24h values of Scr and BUN values were significantly lower compared with the positive perfusion group, the difference was statistically significant (P<0.05); retrograde perfusion group48h after Scr values and BUN values and their differences without statistically significant (P>0.05). Compared with the sham group, the retrograde perfusion group perfusion group in the postoperative6h,12h,24h and48h of CysC values and KIM-1value were higher, the difference was statistically significant (P<0.05). Compared with positive perfusion group, the retrograde perfusion group after6h,12h,24h and48h of CysC values and KIM-1values were significantly reduced, the difference was statistically significant (P<0.05).2.2Renal pathology HE and immunohistochemical results:Pathological sham operation group, no obvious abnormalities, postoperative6h,12h,24h,48h, retrograde perfusion group, perfusion groups had different levels of renal pathological changes:edema of the tubular epithelial cells, lumen, visible protein castsrenal interstitial congestion, and a small amount of inflammatory cell infiltration. In12h,24h, performance of heavier, more gradually ease in the48h after the performance, and the retrograde perfusion group of pathological damage was lighter than positive perfusion group. Sham operation group at6h,12h,24h,48h KIM-1immunohistochemical pathology without significant change. Retrograde perfusion group, is the perfusion group KIM-1labeling index was significantly increased, the difference was statistically significant (P<0.05); compared with positive perfusion group, the retrograde perfusion group KIM-1labeling index was significantly reduced, the difference statistically significant (P<0.05). Retrograde perfusion group KEM-1immunohistochemical pathological damage and gradually repair characteristics and a positive perfusion the group immunohistochemical pathology were shown to strongly positive, the damage was more severe characteristics.[Conclusion]l.Autologous orthotopic liver transplantation operation, via the portal vein method is simple, easy to control the operation time, high success rate, postoperative long survival time after liver transplantation is similar to the pathophysiological changes in the characteristics of the study after liver transplantation organs byloss aspects of an animal model.2.Rat perfusion and retrograde perfusion of autologous liver transplantation on renal both acute injury, the most serious performance after12h and24h to48h after mild renal injury, and retrograde perfusion than perfusion to reduce the ratacute kidney injury. |
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