| Objective:To explore the effects of DAD A on the liver apoptosis index and the expressions of Cytochrome C,FAS and Bcl-2 in rats with severe acute pancreatitis (SAP). Methods:78 rats weight 250-300g were randomized into 4 groups:The normal group (n=6),sham-operation group (n=24), SAP group (n=24)and DADA treatment group(n=24). The model of SAP was established by injection 5% sodium taurocholate 1ml/kg solution into the capsula pancreatis of the rats(Injection velocity 0.2ml/min).The sham-operation group just Flip intestinal wall after laparotomy.Postoperative fasting, free water after awoke. 10min after postoperative DADA group started to give DADA injection (0.357mL/kg/12h) by tail intravenous injection; SAP group use the same dosage of saline.At the 12,24,48 and 72 hour after the establishment of models kill corresponding Four groups rats.AST,ALT, TBIL, DBIL IBIL, AMY were determined. The livers were drawn for light and electron microscopic observation, graded pathology score of liver tissue. Liver apoptosis index was determined by TUNEL method. The expression of Cytochrome C,Fas and Bcl-2 were located by immunohistochemical staining method of SABC. Results:1,pancreas tissue pathology change:(1) In general, there were not obviousiy change in rat pancreas of sham-operation group in different time points. SAP 12hour's group ascites obviouiy, pancreas present dark purple color, hyperemia and edema obviouly, local necrosis, bleeding; 24 hours' there were more ascites surrounding the pancreas, pancreas presents beige, adhesion With the surrounding visceral part, can see large necrotic and bleeding; 48 hours'there were large of hemorrhagic ascites, pancreas presents beige, with large adhesion and viscera necrosis around, hemorrhage; 72 hours' rats were more serious, large effluvial hemorrhagic ascites in Intra-abdominal, dilated bowel seriouly silting bulge, pancreas present beige, with large adhesion with the surrounding viscera, can see extensive calcification, hemorrhage, necrosis.DADA group still can see ascites, pancreas present dark purple color with surrounding viscera's adhesion, flake necrosis, bleeding, but reduce significantly. (2)pancreas morphologic changes in Light-microscopy:each time point of sham-operation group the whole structure of rats pancreatic tissue can be see and there were no inflammatory cell Infiltrates.12 hour's SAP group rats'pancreas leaf interval, flocculus acinar interval clearance, widened, the leaflets structure disorder, a lot of inflammatory cell, pancreatic acinar cells swelling, have focal necrosis, hemorrhage; 24 hours'can see part of normal structure of flocculus disappear, interstitial of inflammatory cells infiltrating, pancreatic acinar cells serious swelling, a flake necrosis, hemorrhage; 48 hours'lobular normal structure disappearr, interstitial of inflammatory cells infiltrating, pancreatic acinar cells extreme swelling, visible extensive necrosis, hemorrhage; 72 hours'normal lobular structure is disappear, interstitial of inflammatory cells infiltrating, pancreatic acinar cells extreme swelling, extensive necrosis, hemorrhage, acinar structures disappear in necrosis area, nuclei is dissolved, can see calcification. DADA group rats' pancreas, still can see lobular structure disorder inflammatory cell infiltrating,pancreatic acinar cells swelling, necrosis, bleeding but reduced significantly.2. Liver tissue pathological changes and histopathological score:(1)each time point of sham-operation group, there were no abnormalities in General form of rat liver.12 hours' SAP group liver bulk increase slightly, coated normal; 24 hours' liver bulk slightly increase, coated normal, and a little adhesion with the pancreas; 48 hours' liver bulk increase slightly, coated nervous, slightly pale, and adhesion with pancreatic obviously, liver tissue necrosis can be seen in adhesion areas; 72 hours'liver bulk increase slightly, coated nervous, color becomes weak obviously, and adhesion with pancreatic were more heavier, liver tissue necrosis obviously in adhesion areas with pancreas. There were not obviously difference in general from between 12hours' DADA group and SAP group; 24 hours'liver bulk increase slightly; 48 hours'liver bulk increase slightly, coated nervous slightly, minority adhesion with pancreas and no liver necrosis.72 hours' liver bulk increase slightly, coated slightly nervous, adhesion with pancreas slightly, liver necrosis notable. (2)liver tissue morphologic changes in Light-microscopy:each time point of sham-operation group, there were no abnormalities in liver tissue.12hours'SAP group liver cell mild edema, cellular gap widened; 24 hours'liver swelling increased,and with more inflammatory cells infiltrating, fat change, a few of liver tissue can see Erythrocytes deposition; 48 hours'liver visible focal or irregular sheets necrosis, liver sinus and collect abbacy of inflammatory cells infiltration and spread out red blood cells around; 72 hours' larger area of liver visible focal or irregular sheets necrosis, liver sinus and collect abbacy of more inflammatory cells infiltration and spread out red blood cells around; Each DAD A group still can see liver cells swelling, inflammatory cells infiltrating and area of liver cells necrosis, but compere with SAP group the degree is decreased. (3)Histopathological examination score:12 hours' liver tissue pathology score SAP group were hinger than sham-operation group (P< 0.05), DADA group and SAP group get not obvious difference (P> 0.05).24 hours',48 hours',72 hours', SAP group were hingher than sham-operation group (P< 0.05)and DADA group were lower than SAP group respectively (P< 0.05).72 hour reache the peak.4,AMY,AST et al index expression:sham-operation groups without apparent change in different time points; SAP group and DADA group is obviously increased in different time points compered with sham-operation group (P < 0.05), DADA group were lower than SAP group (P< 0.05).5,Liver cell apoptosis index, Fas, Cytochromes c and Bcl-2 gene expression: sham-operation without obvious change in different time points, SAP group and DADA group were increased obviously compered with sham-operation group in different time points (P< 0.05), DADA group were lower than SAP group (P< 0.05). The Bcl-2 gene expression without obviously change in sham-operation group in different time points, SAP group and DADA group the Bcl-2 gene expression were reduced significantly than sham-operation group in different time points (P< 0.05), DADA group were higher than SAP group (P< 0.05). Conclusion:1. DADA have protectie effect to rat pancreas induced liver damage in acute pancreatitis,it may provides a new way of pancreas induced liver injury treatment for clinical.2.Liver cell apoptosis plays an important role in the pathogenesis of SAP liver injury, and apoptosis related factors of Bcl-2,Fas plays a regulation function in liver cell's apoptosis.3. In early stage of SAP, patients use DADA treatment can reduce the express of poptosis related factors Fas, increase Bcl-2 expression, improve liver cell mitochondrial function thus inhibiting liver cell apoptosis excessively to reduce liver injury degree and good for improve the prognosis.
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