Protective Effect And Mechanisms Of RhUTI On Experimental Panctreatitis Animal Models

ObjectiveTo study the protective effect and mechanisms of rhUTI on experimental pancreatitis animal models, and to provide the theoretical basis for reporting national first-class drug and clinical applications.Methods1. The rats model of acute pancreatitis were established by injecting 2% sodium taurocholate (0.1mL/100g, 0.2mL/min) into the pancreaticobiliary duct. Acute pancreatitis rats were randomly divided into six groups: cntrol group, model group, rhUTI (17,500,35,000,70,000U/kg) groups and UTI(35,000 U/kg) group, ten rats in each group. After experiment, the changes of serum amylase,urine amylase,complete blood count,urea nitrogen,creatinine,K+,Na+,Cl-,Ca2+ and pathologic histology of pancreas of acute pancreatitis rats were detected.2. The dogs model of acute pancreatitis were established by injecting 2.5% sodium taurocholate (1ml/kg) into the pancreaticobiliary duct. Acute pancreatitis dogs were randomly divided into six groups: cntrol group, model group, rhUTI (7,500,15,000,30,000 U/kg) groups and UTI(15,000 U/kg) group, six dogs in each group. After experiment, the changes of serum amylase,urine amylase,complete blood count,urea nitrogen,creatinine,K+,Na+,Cl-,Ca2+ and pathologic histology of pancreas of acute pancreatitis dogs were detected.3. The distribution of apoptotic acinar cells of acute pancratitis rats was observed by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The protein and mRNA expressions of Fas, Fas-L and Caspase-3 in pancreatic tissue were measured by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR) of acute pancreatitis rats.Results1. Rats model of acute pancreatitis was successfully prepared by retroinjecting 2% sodium taurocholate at dose of 0.1mL/100g body weight and at speed of 0.2mL/min into the pancreaticobiliary duct. Compared with control group, the levels of serum amylase,urine amylase,red blood cell count,urea nitrogen and creatinine raise, while the level of Ca2+ descended, and the pancreatic tissue were hyperemia,oedema,focal necrosis and infiltrated with inflammatory cells. Compared with AP group, the levels of serum amylase and urine amylase descended in UTI (35,000U/kg) group and rhUTI (35,000,70,000U/kg) groups, the extent of hyperemia,oedema,necrosis and inflammatory infiltration of pancreatic tissue were lessened among UTI and rhUTI groups. However, there were no obvious effects on complete blood count,urea nitrogen,creatinine and electrolyte among UTI and rhUTI groups.2. Dogs model of acute pancreatitis was successfully prepared by retroinjecting 2.5% sodium taurocholate at dose of 1mL/kg body weight into the pancreaticobiliary duct. Compared with control group, the level of serum amylase raise after 24h,28h and 32h, and reached a peak at 32h, the level of urine amylase raise after 32h, the level of serum K+ and Ca2+ descended after 24h and 8h respectively and the pancreatic tissue were hyperemia,oedema,necrosis and infiltrated with inflammatory cells. However, there were no obvious changes about complete blood count,urea nitrogen and creatinine in AP group. Compared with AP group, the level of serum amylase descended after 28h and 32 hour in UTI and rhUTI groups, the level of urine amylase descended in rhUTI (15,000,30,000 U/kg) groups and UTI(15,000U/kg) group and the extent of hyperemia,oedema,necrosis and inflammatory infiltration of pancreatic tissue were lessened among UTI and rhUTI groups.3. Tunel assay showed that minimal apoptosis in normal pancreatic acinar cells of rats. Compared with control group, the pancreatic acinar cell apoptosis index increased significantly in AP rats. Because of severe pathological damage, there appeared rare apoptotic cells in model group, while compared with AP group, the apoptotic cells increased significantly in UTI and rhUIT groups. In normal pancreatic tissue of rats, there appeared the protein and mRNA of Fas,Fas-L and Caspase-3. Compared with control group, the expression of protein and mRNA of Fas,Fas-L and Caspase-3 descended; while compared with AP group the expression of above protein and mRNA raise significantly in UTI and rhUTI groups, and the tendency of changes among Caspase -3 and Fas,Fas-L were coincidence.ConclusionsrhUTI may have protective effect on acute pancreatitis animals,it can inhibite amylase activity and improve pathological damage of pancreatic. Meanwhile the mechanisms might be related with advancing pancreatic acinar cell apoptosis and upregulating the expression of the protein and mRNA of Fas,Fas-L and Caspase-3 in pancreatic tissue.