Experimental Study On The Expression Of Caspase-3 And Survivin In Rats With Acute Pancreatitis

Objectives To establish acute pancreatitis(AP) animal model and observe the expression of Caspase-3 and Survivin in the different groups of acute pancreatitis. To observe the changes of apoptosis factors and theirs effects on the body in pancreatic tissue of experimental acute pancreatitis. To explore the role of Caspase-3 and Survivin in acute pancreatitis and theirs relationship with the damage of pancreatic tissue. To discuss the pathogenesis and development mechanism of the AP from the point of the pancreatic cell apoptosis. This studying can provide a science basis by judging and predicting the outcome of AP.Methods The specific pathogen free(SPF) wistar rats were randomly divided into four groups: sham-operated group, six hours after AP group, twelve hours after AP group, twenty-four hours after AP group. AP animal model was induced by injection of 5% Na-Tc under the membrane of pancreas. The sham-operated group rats were killed at 6th hour after operated. The 6h after AP group were killed at 6th hour after operated. The 12 h after AP group were killed at 12 th hour after operated. The 24 h after AP group were killed at 24 th hour after operated. Each experimental rats were extracted 3 ml blood from abdominal aortic. We tested serum amylase(AMY) by appling fully automatic biochemical instrument and taked the pancreatic tissues with HE staining for pancreatic histopathologic scores. We observed the expression of Caspase-3 and Survivin in pancreatic tissue by appling immuno-histochemical method.We analyzed all the data by used SPSS 18.0. All the data were shown as means ± standard deviation( x ± s). We compared the difference of the groups by One-way ANOVA method. We measure the correlationship of AMY, pancreatic histopathologic score, Caspase-3, Survivin by Pearson correlation analysis. The inspection standards was alpha = 0.05.Results We established the AP animal model triumphantly. The rats of AP group exhibited the high amylase and the pancreatic pathological injury. The pancreas pathological damage increased gradually as time goes on. The content of serum amylase in sham-operated group compared with 6h after AP group was significant difference(P<0.05), The content of serum amylase in sham-operated group compared with 12 h after AP group was significant difference(P<0.05), The content of serum amylase in sham-operated group compared with 24 h after AP group was significant difference(P<0.05). The content of serum amylase in 6h after AP group, 12 h after AP group, 24 h after AP group compared with each other were significant difference(P<0.05). The pancreatic histopathologic scores in sham-operated group compared with 6h after AP group was significant difference(P<0.05), The pancreatic histopathologic scores in sham-operated group compared with 12 h after AP group was significant difference(P<0.05), The pancreatic histopathologic scores in sham-operated group compared with 24 h after AP group was significant difference(P<0.05). The pancreatic histopathologic scores in 6h after AP group, 12 h after AP group, 24 h after AP group compared with each other were significant difference(P<0.05). The optical density(OD) of Caspase-3 in sham-operated group compared with 6h after AP group was significant difference(P<0.05), The optical density(OD) of Caspase-3 in sham-operated group compared with 12 h after AP group was significant difference(P<0.05), The optical density(OD) of Caspase-3 in sham-operated group compared with 24 h after AP group was significant difference(P<0.05). The optical density(OD) of Caspase-3 in 6h after AP group, 12 h after AP group, 24 h after AP group compared with each other were significant difference(P<0.05). The optical density(OD) of Survivin in sham-operated group compared with 6h after AP group was significant difference(P<0.05), The optical density(OD) of Survivin in sham-operated group compared with 12 h after AP group was significant difference(P<0.05), The optical density(OD) of Survivin in sham-operated group compared with 24 h after AP group was significant difference(P<0.05). The optical density(OD) of Survivin in 6h after AP group, 12 h after AP group, 24 h after AP group compared with each other were significant difference(P<0.05). The Caspase-3 was highly negative correlation(r=-0.686, P<0.05) with Survivin. The Caspase-3 was highly negative correlation(r=-0.800, P<0.05) with amylase. The Caspase-3 was highly negative correlation(r=-0.802, P<0.05) with pancreatic histopathologic scores. The Survivin was highly positive correlation(r=0.811, P<0.05) with amylase. The Survivin was highly positive correlation(r=0.821, P<0.05) with pancreatic histopathologic scores. The amylase was highly positive correlation(r=0.950, P<0.05) with pancreatic histopathologic scores.Conclusions There were disturbances function of apoptosis system in the early stage of acute pancreatitis. And the disturbances of Caspase-3 and Survivin were aggravated as the illness grew worse. There was a positive correlation between apoptosis factors Caspase-3, Survivin and the pancreatic histopathologic scores, serum amylase. These changes can be considered as the experiment indexes which can predict the severity of severe acute pancreatitis.