Research Of Mechanism Of Effect Of Cyclooxygenase -2 Inhibitors On SAP Rats Mode

Objective:Acute pancreatitis (acute pancreatitis, AP) usually presents arising fast, destroing severily, changing quickly, being more complications, especially severe acute pancreatitis (severe acute pancreatitis,SAP) is potential risks of high mortality rate. The occurrence of this disease is relation with pro-inflammatory cytokines and inhibitors in the imbalance on the network. But its pathogenesis is not clear,and the disease is a lack of specific and effective drug treatment. Recent studies have foucsed the occurrence and development in SAP signaling pathway in the interaction of various cytokines, resulting in a large variety of inflammatory factors, and thus result in systemic inflammatory response syndrome, ultimately the reaction can lead to multiple organ failure.How to effectively regulate the production of cytokines,how to prevent the occurrence of SIRS or reduce its severity, and how to enhance the therapeutic effect of SAP and reduce the mortality became the a huge problem in front of the medical scientist. Studing the effectiveness of cyclooxygenase-2 inhibitor, how to regulating cell factor (TNF-a, IL-6 and IL-10) production, how to prevent the occurrence of SIRS or reduce its severity, and how to enhance the therapeutic effect of SAP and reduce deaths rate is hot point. This subject using sodium taurocholate in rats with severe acute pancreatitis model about the pathway study to examine the mechanism of effect of cox-2 inhibitor on SAP rats model. Methods:45 adult male SD rats were randomly divided into three groups:sham operation group (SO group), SAP Group and SAP-Celecoxib group (SC group). 3h,6h,12h three time points, each time points 5. The model of SAP was induced by the retrograde injection of 5% sodium taurocholate (1 ml/kg) in the bile-pancreatic duct. Rats did not be bred before surgery but could be watered. Injected 3% of the sodium pentobarbital(1ml/kg) fulfill intraperitoneal anesthesis. Routine disinfection of shop towels, upper abdominal midline incision 2cm. Use of 1ml bosch injector blunt needle puncture in the duodenal descending part near the lateral wall,from the duodenal end of pancreatic duct retrograde penetrate about 1cm, respectively, near the liver hilum with noninvasive pancreatic duct clipping, into the 5% Tc-Na (injection rate 0.1ml/min).5min later, check and confirm that the rat, s pancreas edema and then push the needle out of the pancreatic duct, release the artery clip, confirmed no active bleeding in the abdominal abdomen, finally close the incision with two-tier, and inject the normal saline (2ml/100g) in the back of subcutaneous to supplement the water.SO group deal with the SAP group, but the pancreatic duct inject the normal saline. SC in the surgery group were 0.5% Celecoxib preparation of fresh solution to 50 mg/kg body weight dose gavage 2 hours before the induction of SAP, the follow-up operation with the SAP group. The model group was not successful the first puncture and intraoperative death eliminated.15 rats in each group sub-3h,6h,12h three time points, each time point five rat of abdominal examination. To observe the changes in abdominal pancreas, ascites measured changes cardiac blood of about 3～5ml, extracted serum, serum samples stored at-200C, and prepare for follow-up testing. Pancreatic tissue obtained with 4% paraformaldehyde and immersion fixed 24h, embedded in paraffin, sliced. After the rats bleed to death lung tissue continuous baking oven set 800C 24h, to calculate the wet/dry ratio.Automatic biochemical analyzer serum amylase and serum by ELISA, TNF-α, IL-6, IL-10 and other cytokines.Naked eye pancreas, lung hemorrhage, edema and other changes; HE staining, light microscopy of pancreatic tissue pathology.Results:1. Ascites:SO group had no bloody ascites; SAP group, SC group had bloody ascites, SC group at each time point the amount of ascites than SAP group was statistically significant (p=0.008).2. Serum amylase:SO low serum amylase activity at different time points were lower than the other two groups (p=0.005); SC group level in each group were decreased compared with SAP, but only in 6h,12h was statistically significant (p=0.002).3. Serum cytokines: SO serum TNF-a, IL-6 and IL-10 were significantly lower than the SAP group and the SC group was statistically significant (p<0.01). SC group TNF-a, IL-6 and IL-10 was lower than SAP group, TNF-a, IL-10 at each time point were statistically significant (p<0.05); IL-6 at 6h,12h were statistically significance (p<0.05).4. Lung wet/dry weight ratio:SO group at each time point ratio was lower than SAP group and the SC group was statistically significant (P=0.034); SC ratio of less than SAP group at each time group,6h,12h statistically significance (P=0.023).5. Pancreas generally score:SO eye view of no change; SAP group at each time point were significantly higher pathological scores; SC group than the SAP group scores decreased in the 6,12-hour difference was statistically significant (p=0.006).6. Pancreatic histological score:SO obvious pathological changes of pancreas, SAP in each group were pancreatic interstitial infiltration of inflammatory cells and red blood cell leakage, and increased with the degree of disease progression.3h group see a small amount of pancreatic interstitial inflammatory cell infiltration, acinar cells a small amount of necrosis; 6h large number of red blood cells of pancreas interstitial exudation and inflammatory cell infiltration, acinar cell necrosis in a wide range of severe swelling; 12h group necrosis was expanded up to the leaflets, but still see the outline of leaflets; CB treatment group also showed obvious pathological changes in the pancreas, but more to reduce SAP group, the two groups at each time point were significantly different (p=0.003).Conclusion:1. Cytokines TNF-a, IL-6 and IL-10 expression levels and the severity of SAP is related to the development of the disease in the SAP plays an important role.2. Cyclooxygenase-2 inhibitor reduced TNF-α, IL-6 and IL-10 expression and decreased the expression of inflammatory cytokines,improved the pathological pancreas and lung injury. The research point out that Cyclooxygenase-2 inhibitors in the pathogenesis of SAP rats plays an important role in the pathway as therapeutic target for SAP, Inhibitors control its SAP application value, but the application of Celecoxib in the SAP before the onset of 2 hours using, the stronger its preventive treatment of pancreatitis significance.