| Objective: To establish an ideal animal mode with severe acute pancreatitis(SAP).To observe the expression of nuclear factorκB(NF-κB) in spleen tissue in the rats with SAP, and investigate the role of spleen in the pathogenesis of SAP. Methods: 1) SD rats were randomly divided into experimental group (SAP,n=24) and sham operation group(n=24), The SAP model was induced by retrograde injection of 4% sodium taurocholate into biliopancreatic ducts with fine needle. The ascites volume was calculated in each group. The serum amylase activity at 3h, 6h and 12h were measured respectively. The tissue sample of pancreas were harvested in each group, serially sectioned after formalin fixation and embedded in paraffin, prepared in 4μm hematoxylin-eosin stained sections, and evaluated histologically under conventional light microscopy. Pathological change of pancreas was evaluated by Rongine score system.2) Ninety-six rats were randomly divided into four groups :sham operation group, splenectomy group, SAP group and SAP with splenectomy group?each contained 24 rats .The rats were sacrificed at 3h,6h and 12h after operation, and the blood samples and tissue samples of each group were gathered for the following detection: change of pancreatic tissue observed by microscope with HE staining and scored by Rogine criteria; Serum tumor necrosis factor-α(TNF-α) was measured by ELISA method; Immunohistochemical method was used to observe the activity of splenic NF-κB p65 in the rats with SAP. Results: 1) The pathological morphological change of edema, hemorrhage, necrosis and inflammatory cells infiltration were observed in pancreatic tissue at 3,6th and 12th after SAP model induction; Pancreatic pathological evaluation, ascites volumes and serum TNF-αof SAP group is significant higher than sham operation group.2)The level of TNF-αin SAP and SAP with splenectomy group is significant higher than sham operation group and splenectomy group in all time point (P<0.05), the level in SAP with splenectomy group was significantly decreased compared with SAP group at 6h and 12h.The level in SAP was gradually elevated from 3h to 12h,and all comparison was significant .(P<0.05).While in SAP with splenectomy group the level at 3h to 6h and 6h to 12h was no statistical difference (P<0.05). Histological examination in the AP with splenectomy group were significantly milder than those in the AP group at 6h(7.83±0.753,vs 9.67±1.211) and 12h(9.67±0.816 , vs13±0.894) (P<0.01), The positive expression of splenic NF-κB p65 began to be intensified at 3 h(46.967±1.148) ,peaked at 6h(56.333±1.588),attenuated at 12h (36.900±0.756) in the rats with AP, and all show insignificant differences compared with the sham operation group(3.400±0.800). Conclusions: SAP model is successfully induced by stable operating plat and retrograde injection of 4% sodium taurocholate into biliopancreatic ducts with fine needle. It minimizes the trauma and simplized the procedures. It might be a reliable model with high success rate and an ideal severe acute pancreatitis animal model for recommendation. Spleen had much effect on the development of SAP, which splenectomy can make acute pancreatitis milder,Spleen may promote the production and release of TNF-αand other inflammatory mediators due to activation of splenic NF-κB in the develop and progression of SAP and it plays important role in the pathogenesis of AP. |
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