| Background and purposeSevere acute pancreatitis(SAP)is a common clinical acute abdomen with a high morbidity and mortality rate [1,2].Previous studies have shown that intestinal barrier dysfunction plays an important role in the progression of SAP [3,4].The damage of intestinal barrier function can lead to the translocation of bacteria and endotoxin into the blood and lymph circulation,can lead to secondary infection of pancreatic tissue,resulting in multiple organ failure(MOF)[5].Thus,the protection of intestinal barrier function has become an important treatment for SAP.However,at present,the main treatment methods,such as early enteral nutrition,the use of probiotics and antibiotics are not ideal.Therefore,to find an effective treatment strategy has become an urgent problem to be solved.As we all know,SAP can produce pancreatitis associated ascitic fluid(PAAF).PAAF has been confirmed to contain a large number of inflammatory mediators such as cytokines,enzymes,which can damage the intestinal barrier function.Therefore,we believe that if the intervention of PAAF is carried as soon as possible,it may improve the cond ition and prognosis of patients with SAP.Based on this,our team use abdominal peritoneal drainage(APD)for drainage of PAAF for patients with SAP.A large number of clinical data of retrospective analysis showed that APD can not only significantly impro ve the prognosis of patients with SAP,but also reduce the mortality rate of patients with SAP [6-8].Subsequently,through animal experiments,we further confirmed that APD can not only significantly improve the survival rate of SAP rats,but also has an effect on SAP associated lung injury and lipid metabolism disorder[8-10].Although APD treatment has made some progress,the understanding of its effective role and mechanism is still relatively preliminary,so further research is needed.For example,does APD play a role in intestinal bacterial translocation? Does APD play a beneficial role in intestinal barrier dysfunction?Methods1.Preparation of the model: first,the SD rats' s intestine were prepared by using antibiotics.Then the rats were feed by the E.coli whose plasmids were labeled with GFP(E.coli marked with Fluorescent Protein,GFP-E.coli)for 3 days.After E.coli were successfully colonized in rats' intestine,the rats were divided into sham operation group(Sham),SAP group and APD group,with 30 rats in each group.SAP rat model was prepared by injecting sodium taurocholate into the pancreatic duct.Preparation for APD: drainage catheter was placed in right lower abdomen after SAP.In the sham operation group,the pancreas was turned off several times after laparotomy.2.The mortality and severity of pancreatitis were observed and measured: 24 hours after the operation,the mortality rate and the amount of ascites were observed.Under sterile conditions all the rats were sampled and then killed.HE staining for the pathological score of pancreas in three groups of rats,ELISA method for the determination of serum C reactive protein(CRP),tumor necrosis factor ?(TNF-?),interleukin ?(IL-1?)level,and serum endotoxin level.3.Detection of bacterial translocation: blood,mesenteric lymph nodes(MLN)and pancreatic tissue were cultured and detected for bacteria with fluorescence microscopy,and the bacterial translocation rate was calculated.4.Detection of intestinal barrier function: HE staining for pathological score of intestinal tissue in three groups of rats;ELASA method for serum levels of intestinal Fatty Acid Binding Protein(i FABP),Diamine Oxidase(DAO)and D-lactid;immunohistochemistry method for the detection of intestinal mucosa tight junction protein zona occludens-1(ZO-1).Results1 Severity of pancreatitis: 1.1 mortality and ascites: no rats died in Sham group,but the mortality rate of APD group was significantly lower than that of SAP group,and the difference was statistically significant(p<0.05).There was no ascites in Sham group,but the ascites volume of APD group was significantly less than that of SAP group,and the difference was statistically significant(p<0.05).1.2 Compared with group Sham group,the pathological score of pancreatic tissue in SAP group and APD group was significantly increased(p < 0.05);while compared with SAP group,pathological scores was significantly lower in APD group(p < 0.05).1.3 Compared with Sham group,serum CRP,TNF-? and IL-1? level in SAP group and APD group increased significantly,and the difference was statistically significant(p < 0.05);while compared with SAP group,serum CRP,TNF-? and IL-1? level in APD group were significantly lower,and the difference was statistically significant(p < 0.05).2 Bacterial translocation: 2.1 In Sham rats,no bacterial translocation was found in MLN,blood and pancreatic tissue.No bacterial translocation was found in all the blood samples in the three group.Bacterial translocation was found in MLN and pancreatic tissue in both SAP group and APD group.While the the translocation rate in APD group was significantly lower than that in SAP group,and the difference was statistically significant(p <0.05).2.2 Compared with Sham group,serum endotoxin level in SAP group and APD group was significantly increased,and the difference was statistically significant(p < 0.05);while compared with SAP group,serum endotoxin level in APD group,was significantly decreased,and the difference was statistically significant(p < 0.05)3 Intestinal barrier function: 3.1 Compared with Sham group,the intestinal pathological score in SAP group and APD group increased significantly(p < 0.05);while compared with group SAP,intestinal pathological score was si gnificantly lower(p < 0.05).3.2 Compared with Sham group,serum i FABP,DAO and D-lactate levels in SAP group and APD group of were significantly increased,and the difference was statistically significant(p < 0.05);while compared with SAP group,serum iFABP,DAO and D-lactate levels in APD group were significantly lower,and the difference was statistically significant(p < 0.05).3.3 Compared with Sham group,the expression of intestinal mucosal tight junction protein ZO-1 in SAP group and APD group decreased significantly,and the difference was statistically significant(p < 0.05);while compared with SAP group,the expression of ZO-1 in APD group increased significantly,and the difference was statistically significant(p< 0.05).Conclusion1.APD significantly reduced the inflammation and mortality rate of SAP.2.APD significantly improved the translocation of intestinal bacterial and endotoxin.3.APD significantly increased the expression of tight junction protein ZO-1 in intestinal mucosa and improved intestinal barrier dysfunction. |
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