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Objective: to establish a stable, physiological small the acute pancreas transplantation model. 2. Observed cryopreservation for pancreatic external secretory function and perfusion. Explore the relationship for of pancreatic apoptosis and cold ischemic injury and Bcl-2, Bax protein in which the role. Method: 1. Allogeneic pig whole pancreatic duodenal transplantation model and evaluation: 30 healthy miniature pigs were randomly divided into for receptor groups, application of 4 ℃ UW solution in situ perfusion donor pig pancreas After the duodenum, the entire cut donor total pancreaticoduodenal. Subsequently, the purposes of the pig allogeneic pancreaticoduodenal transplants. Will move for pancreatic vein and recipient pigs abdominal aorta and inferior vena cava anastomosis, venous flow into the systemic circulation, the duodenal segments receptor jejunum consistent. Intraoperative monitoring receptor heart rate, mean arterial blood pressure, oxygen saturation. After giving conventional rehydration and anticoagulant, anti-infective, observe and evaluate the general situation of the receptor. 2, for the study of pancreatic cold ischemic injury and related mechanisms: 40 healthy miniature pigs were randomly paired purposes of allogeneic pig pancreas transplantation 20. Thereafter, the donor porcine pancreatic duodenal perfusion in situ and cut immediately after placement cryopreservation 4 ℃ UW solution, depending on the storage time were divided into four groups: 1, cold ischemia time (CIT) 0h group : successfully cut the graft did not undergo cold preservation and transplant immediately; 2, 4 ℃ UW solution cold ischemia time (CIT) 3h: graft placement save 3h after transplantation; 3, cold ischemia time (CIT) 6h group : graft placed 4 ℃ UW solution to save 6h after transplantation; 4, cold ischemia time (CIT) 9h: graft placed 4 ℃ UW solution to save 9h after transplant. For this period, porcine pancreatic resection receptor made into a model of diabetes. The pancreas transplant complete the detection for pancreatic blood flow after 30min perfusion and get for pancreatic tissue specimens were divided into three sets, were used as electron microscopy, apoptosis in situ end labeling (TUNEL) and Bcl-2, Bax protein expression determination. In addition, pre-transplant, respectively, after extraction receptor venous blood, measured relative to glucose, blood amylase and serum insulin concentrations. Results: 1, completed in a large animal model of the process of establishing full pancreaticoduodenal transplant 15 cases, 1 patient with intraoperative excessive bleeding after 2h died of circulatory failure; another example of a result of the bowel - intestinal anastomotic fistula in postoperative 3d death; were survival. Surgical procedure for the average operative time (69.3 ± 7.2) min, receptor mean operative time (75.5 ± 13.1) min. 2, four groups for pancreatic blood flow recanalization to 2h after receptor pig blood sugar began to decline and stabilized after 24 hours. In for pancreatic transplantation success 2h, 8h, the CIT 9h blood glucose, blood amylase, blood flow resistance index (RI) were significantly higher than the other three groups (P lt; 0.05), CIT 3h group and CIT 6h group was significantly higher than CIT 0h group (P lt; 0.05), while there was no significant difference between the CIT 3h group with CIT 6h group (P gt; 0.05); abdominal aortic blood flow peak velocity (Vmax) in the CIT 9h group was significantly lower than the other three group (P lt; 0.05), CIT 3h group and CIT 6h group was significantly lower than CIT 0h group (P lt; 0.05), there was no significant difference (P gt between CIT 3h CIT 6h group; 0.05). 3, TUNEL staining found recanalization 0.5h, within each group for pancreatic tissue of pancreatic acinar cells were seen scattered in the distribution of nuclei stained positive for pancreatic blood flow, and the longer cryopreservation for pancreatic unit area The more the number of positive cells. The quantitative analysis of the results found that the extension of the four groups for pancreatic apoptosis index (Apoptosis index, AI) with cold ischemia time, in order: CIT 0h group lt; CIT the 3h group lt; CIT 6h group, lt; CIT 9h group, the differences between the groups were significant (P lt; 0.05). Electron microscopy confirmed for pancreatic perfusion during organizational memory, including the endoplasmic reticulum, mitochondria mild swelling nucleus concentration, different chromatin increased apoptotic acinar cells. In addition, the blood flow through the 0.5 h detected expression of Bcl-2 protein for pancreatic tissue, positive substances are mainly distributed in the cytoplasm and nucleus, quantitative analysis of the four groups, significant differences in the expression of Bcl-2 protein (P lt; 0.05), in which CIT 0h significantly higher than the other three groups (P lt; 0.05), CIT 3h group and CIT 6h group was significantly higher than the CIT 9h group (P lt; 0.05), CIT 3h group with CIT the 6h group showed no significant differences (P gt; 0.05); Bax protein expression were seen within the four groups for pancreatic tissue, positive substances are mainly distributed in the cytoplasm. Which CIT 9h group was significantly higher than the other three groups (P lt; 0.05), CIT 3h group and CIT 6h group was significantly higher than the CIT 0h group (P lt; 0.05), CIT 3h group with CIT 6h group showed no significant differences (P gt; 0.05). Conclusion: 1, the use of allogeneic pancreatic transplantation model miniature pigs for experimental animals, the establishment of the pig, the surgery will be for pancreatic artery and vein, respectively, with the recipient abdominal aorta and inferior vena cava anastomosis, duodenal segment receptor jejunostomy. The model is simple, stable, reliable, and may well be a more ideal model to study pancreas transplantation. 2, cold ischemic injury pancreas the pancreas transplantation process for one of the important reasons for transplantation of pancreatic insufficiency and hypoperfusion. Therefore, if we can reduce the extent of the injury for pancreatic cold ischemia, may have an important role in the promotion for the restoration of the function of the pancreas to increase the amount of pancreatic perfusion and improve the success rate of pancreas transplantation. 3, another form of cell death - apoptosis involved for pancreatic cold preservation injury process, and with the extension of the cold ischemic time, increased the degree of apoptosis of cells in the pancreas. Therefore, if we take effective measures to inhibit apoptosis, the degree of injury may be saved to reduce the low temperatures for pancreatic the improved postoperative for pancreatic function is important. 4, Bcl-2, Bax protein involved in for of pancreatic cold ischemic injury process, in which the expression of Bcl-2 protein inhibited for pancreatic apoptosis; Bax protein expression can promote apoptosis. Addition not only Bcl-2 and Bax protein expression degree for pancreatic apoptosis of Bcl-2/Bax ratio for pancreatic apoptosis was significantly negative correlation. Thus, induction of Bcl-2 expression or inhibition of the expression of Bax promote the Bcl-2/Bax ratio increases is likely to suppress for pancreatic bubble apoptosis.
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