| [Objective]1. To explore the establishment and application of the models of autologous orthotopic liver transplantation in rats with the inferior vena cava retrograde perfusion and via the portal vein perfusion.2. To compare the effect on acute lung injury in rats after the inferior vena cava retrograde perfusion and via the portal vein perfusion in autologous orthotopic liver transplantation.[Methods]1. Experiment one:SD rats were randomly divided into retrograde perfusion group, perfusion group and sham operation group. After the portal vein, the suprahepatic vena cava and infrahepatic vena cava were blocked, the fluid poured into the portal vein, through the infrahepatic vena cava outflow lavage liver, until after liver had been lavage into khaki, and then turn to lift the vessel occlusion to restore liver blood for complete in orthotopic autologous orthotopic liver transplantation. When lift the vessel occlusion, the perfusion group opened the portal vein first, in order to make the blood in portal vein perfusion liver first, and then opened the inferior vena cava; the retrograde perfusion group opened inferior vena cava first, to make the blood in inferior vena cava perfusion liver first, and then opened the portal vein. The sham operation group were only separated out the vein, inferior vena cava, and no vascular occlusion and lavage liver. After models were established, calculated operation time and survival rate, the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin(TB) were measured in12th hours, one day, three days and seven days after operation. The rats were randomly selected in each group and were killed in12th hours and7days after operation and the liver pathological section were observed. 2. Experiment two:SD rats were randomly divided into retrograde perfusion group, perfusion group and sham operation group. To establish the same models with experimental one, The rats were randomly selected in each group and were killed in6th hours,12th hours and24th hours after operation, and the blood gas, ratio of lung dry-to-wet weight, MPO activity of lung tissue, MDA, TNF-a level were measured. Furthermore, the lung pathological section were observed.3. Statistical analysis:The data were analyzed through SPSS13.0software. Measurement data expressed as mean±standard deviation. Statistical methods using one-way ANOVA+SNK, and t-test. P<0.05was considered statistically significant.[Results]1. To establish the model of rat retrograde perfusion and perfusion autologous orthotopic liver transplantation.The operative time of retrograde perfusion group, perfusion group and sham operation group respectively were (39.00±2.50)min,(39.50±2.00)min and (39.00±1.00)min, and the successful rates of them respectively were93.3%(14/15),83.7%(13/15) and100%(15/15). Compare with the sham operation group:the level of ALT, AST and TB of retrograde perfusion group and perfusion group in12th hours, one day and three days after operation were significantly increased, the difference were statistically significan(P<0.05); seven days after operation, the differences among the three groups were not statistically significant. Compare with the perfusion group:the level of ALT, AST and TB of retrograde perfusion group in12th hours and one day after operation were significantly increased, the difference were statistically significant(P<0.05). Compare in a group:the level of ALT, AST and TB of retrograde perfusion group and perfusion group in12th hours after operation were highest, the difference were statistically significant (P<0.05). The HE staining of liver issue:12th hours after operation, retrograde perfusion group and perfusion group, liver tissue in some extent had the performances:liver cells arranged in irregular, irregular shape, edema degeneration, sinusoidal expansion, central venous expansion, portal area interlobular venous expansion, congestion, inflammatory cell infiltration. And the sham operation group was normal. Seven days after operation, three groups all showed no obvious abnormal.2. The effect on acute lung injury in rats after retrograde perfusion in autologous orthotopic liver transplantationCompared with perfusion group:the value of PH, PaO2, the ratio of lung dry-to-wet weight were lower in retrograde perfusion group, however, the activity of MPO and content of MDA were higher, the difference were statistically significant (P<0.05).24th hours after operation, the above indicators were no significant difference between two groups (P<0.05). The content of TNF-a, the retrograde perfusion group was lower in6th,12th and24th hours after operation, the difference were statistically significan(P<0.05). The HE staining of lung issue:The retrograde perfusion group and perfusion group had different levels of performances:Destruction of alveolar structure, the expansion of the alveolar space, alveolar fusion, alveolar epithelial hyperplasia, hypertrophy, alveolar septal thickening, small vein and capillary were congestion and congestion, and the inflammatory cells infiltration. The performances in6th hours and12th hours after operation were heavier than24th hours. The result of lung pathological injury also showed the performance of the retrograde perfusion group was light er than the perfusion group.[Conclusions]1. The model of rats retrograde perfusion and perfusion autologous orthotopic liver transplantation through portal venous perfusion of liver was a simple operation and with short operative time, high success rate and a long survive time model. At the same time, according to the method of model-making and the changes of postoperative liver function and liver pathological, it could speculated that they were with a similar postoperative changes to allogeneic liver transplantation. Both two models can be used as an ideal animal model to study ischemia and reperfusion injury, the damage of organ after liver transplantation, but not reflect the immune rejection.2. The acute lung injury happened in the early period both after the inferior vena cava retrograde perfusion and via the portal vein perfusion autologous orthotopic liver transplantation in rats. The serious performance at6th hours and12th hours and less lung injury at24th hours after operation. The retrograde perfusion had less effect on acute lung injury than perfusion after operation. This experiment provides some theoretical experimental basis for clinical application of retrograde perfusion liver transplantation... |
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