| Objective:Severe acute pancreatitis(AP)in intestinal mucosa vulnerable to invasion,causes intestinal barrier dysfunction and the intestinal flora shift,shift of microbes and their metabolic products will affect autophagy activity of the pancreas,increase of pancreas injury.Basal pancreatic autophagy maintains homeostasis and metabolism by degrading cells and organelles damaged by aging.Impaired autophagy can lead to pancreatic injury.Clostridium butyricum(CB),as a kind of anaerobic bacteria parasitic in the host intestine and in longterm symbiosis with the host,has strong tolerance to acids,alkalis,high temperature,antibiotics,etc.,and can up-regulate probiotics and down-regulate pathogenic bacteria.Its metabolite butyrate can help repair intestinal epithelial mucosa and inhibit intestinal permeability.It can also reduce the colonization of opportunistic pathogens such as Escherichia coli and Enterococcus by creating an acidic and anoxic intestinal environment,thus reducing the damage of intestinal tract and pancreas.The purpose of this study was to explore the role and possible mechanism of Clostridium caseinate on pancreatic autophagy in severe acute pancreatitis.Methods:1.Establishment of animal model: 32 SPF Sprague-Dawley rats were divided into 4groups by random number table method:(1)Control group(CON group): 6 rats.After 10 days of normal saline administration,the abdominal cavity was turned and the abdomen was closed.(2)Control group(NS group): 6 subjects were given Normal saline intragastric administration 10 days later,normal saline was injected retrograde along the pancreatic bile duct to establish control group.(3)Acute pancreatitis group(Acute pancreatitis group,AP group): 10,saline lavage,10 days with 5% along the retrograde pancreatic and taurocholic acid sodium injection model;(4)Acute pancreatitis Clostridium butyricum intervention group(AP+ Clostridium butyricum group,AP+CB group): 10 individuals.10 days after intragastric administration of clostridium butyricum suspension,modeling is conducted by retrograde injection of 5% sodium taurocholate along the pancreatic bile duct.2.Materials: 24 h after based modeling,open before regular disinfection of the skin,iodine volts,step by step along the incision after open the abdominal cavity,extend the incision,take big abdomen aortic blood,the pancreas,ileum and cecum feces organization.3.The test index: according to the postoperative state of the rat and the general expression of the pancreas,combined with the pathological pathology of the blood amylase and the pancreas to evaluate the condition of the pancreas injury;According to the general performance of the intestinal tract,the contents of the endotoxin,the pathologic,the high and the depth of the hidden area were evaluated to assess the damage of the intestinal mucosa.The size and number of autophagy of the pancreas were observed in the transmission electron microscope to assess the condition of the pancreas.The immunoadsorption method was used to detect the blood and the blood of the blood tnf-4.The expression of lc3 and p62 in pancreatic acinar cells was detected by immunohistochemical method.The number and structure of the intestinal flora were detected by 16 s rRNA.Results:1.Blood amylase content in AP group was significantly increased compared with that in CON and NS groups(P <0.05),and pancreatic damage was obvious under light microscope,indicating successful modeling.After Clostridium caseinate treatment,the damage in AP+CB group was significantly reduced compared with AP group(P <0.05),while the damage in CON group was more severe than that in AP group(P <0.05).2.Blood ET content in AP group was significantly increased compared with CON group and NS group,ileum damage was obvious under light microscope,ileum villus height was significantly decreased(P <0.05),after Clostridium butyrate treatment,ET content in AP+CB group was significantly decreased compared with AP group(P <0.05),villus height was significantly increased(P <0.05).3.The contents of TNF-α and IL-6 in blood of AP group were significantly increased compared with CON and NS groups(P <0.05),and after Clostridium butyrate treatment,the contents in AP+CB group were significantly decreased compared with AP group(P <0.05).4.Blood IL-1β content in AP group was significantly lower than that in CON and NS groups(P <0.05),and after Clostridium butyrate treatment,blood IL-1β content in AP+CB group was significantly higher than that in AP group(P <0.05).5.LC3 and p62 contents in AP group were significantly increased compared with CON group and NS group(P <0.05).After Clostridium butyrate treatment,the contents in AP+CB group were significantly decreased compared with AP group(P <0.05).6.Under electron microscopy,the pancreatic autophagic vesicles in AP group were significantly increased compared with CON and NS groups(P <0.05),and after Clostridium butyrate treatment,the pancreatic autophagic vesicles in AP+CB group were significantly decreased compared with AP group(P <0.05).7.16 s rRNA found that after Clostridium caseate treatment,pathogenic bacteria in AP+CB group were significantly reduced compared with AP group(P <0.05),and probiotics were significantly increased(P <0.05).Conclusion:1.Compared with the AP group,the administration of Clostridium caseinate increased the abundance of intestinal flora,increased the diversity of intestinal flora,increased the beneficial flora,decreased the harmful flora,reduced the damage of ileum intestinal mucosa,reduced intestinal permeability,and increased the body weight of rats.Considering that Clostridium caseinate could protect the intestine from damage by regulating the structure of intestinal flora.2.AP,ET content increased,inflammatory factor TNF alpha and beta proinflammatory factor,such as a large number of release,IL-IL-10 suppression of inflammation factors such as reduced,resulting in the pancreas and intestinal mucosa has a different degree of damage,the intervention of the probiotic butyric acid bacteria can inhibit the release of proinflammatory medium to some extent,by increasing the release of inflammatory factor suppression to alleviate the pancreas injury.3.AP,visible LC3 pancreas and p62 levels,autophagic vacuoles increased,after the intervention,butyric acid bacteria can reverse autophagy injury to some extent,can consider butyric acid bacteria by regulating intestinal flora enhance pancreas autophagy. |
PDF Full Text Request