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Objective: To establish a rat model of severe acute pancreatitis, and explore of Chinese medicine Qingyitang particles of severe acute pancreatitis (severe acute pancreatitis, SAP) barrier function of the intestinal mucosa and its mechanism in rats: select healthy male Wistar rats were divided normal group, sham group and models A, B, C group, the sodium taurocholate cholangiopancreatography retrograde injection method to establish SAP animal models, sham group and models A, B and C rats bile pancreatic duct intubation respectively retrograde injection of saline and 2%, 3.5%, 5% sodium taurocholate, observed mortality and the degree of pancreatic injury; stable SAP rat model selected 150 healthy male Wistar rats were randomly divided 3 groups were sham group, model group and treatment group. Model group and treatment group biliopancreatic duct retrograde injection of 3.5% sodium taurocholate SAP model, before modeling 2h, 6h after modeling, 18h, 30h and 42h were given Qingyi particles gavage treatment group, model In the same way, the group and the sham group respectively give Chinese medicine placebo saline. Animals in each group, respectively, in the modeling, 6h after modeling, 12h, 24h, 48h abdominal aorta blood line detection of amylase and diamine oxidase, pancreatic tissues and terminal ileum HE staining and pathological score simultaneously measure intestinal villus height, mucosal thickness, and applying immunohistochemical methods to detect small intestinal mucosa tight junction protein Claudin-1 expression. Results: 1. Sham group no death records, model A, B, C group after 72h mortality rates were 20%, 50%, 70%; model group B amylase modeling 6h up to 9023.0 ± 1924.5 U / L, after gradually over time decline in 72h reduced to 2245.3 ± 271.6 is still higher than the sham group and the normal group and was statistically significant (p lt; 0.05); endoscopic findings model group B pancreatic interstitial edema; erythrocyte leakage; inflammatory cell infiltration. Acinar cell edema; dot sheet bleeding; focal degeneration and necrosis; part of the catheter dilation. The Pathology Scoring Model B group compared with the sham group at each time point 9.89 ± 2.11 vs 1.27 ± 1.13,10.25 ± 1.98 vs 1.32 ± 1.28,11.71 ± 2.11 vs 1.03 ± 0.11,13.85 ± 1.05 vs 1.01 ± 0.19,13.89 ± 0.89 vs 0.98 ± 0.07 are significant differences. The 2 model group rats spirit dispirited, piloerection, less dynamic tired lying; the treated rats spirit state better, more activities, increased frequency of defecation, perianal pollution. The sham group No deaths recorded low mortality rate of the treatment group SAP rats. Model and the treatment group, 6h, 12h, 24h, 48h mortality was 0% (0/10), 20% (2/10), 40% (4/10), 50% (5/10) and 0% (0/10), 20% (2/10), 30% (3/10), 30% (3/10); treatment groups 24h mortality of less than in the model (P lt; 0.05) 3. sham group starch 6h a transient increase in the enzymes in the model, and subsequently returned to normal; the model group amylase modeling 6h highest, later declined gradually over time, but still higher than in the 48h sham group (p lt; 0.05); with model In comparison, the treatment group after modeling 12h reduce began to have a statistically significant (p lt; 0.05), 48h no difference compared with the sham group. sham group serum DAO levels before and after the modeling did not change (p gt; 0.05); treatment serum levels of DAO modeling after 24 h and 48 h compared with the model group decreased a significant difference (p lt; 0.05) 5. sham group after 6h visible pancreatic acinar cells with mild edema, no bleeding and necrosis. Model were observed in pancreatic interstitial edema, red blood cell leakage, inflammatory cell infiltration, acinar cells the edema, point haemorrhage, focal degeneration and necrosis, part of the pancreatic duct dilation, pancreatic histopathology score gradually increased over time. Treatment group than 24h change to alleviate the pathology score decrease was statistically significant. Light microscope sham group structural integrity of the intestinal mucosal villi, the epithelium was tall columnar, interstitial edema, no neutrophil infiltration. Intestinal mucosa of the model group, significant structural damage and showed progressive changes, especially in molding 24h is the most obvious, visible the rat ileum interstitial edema and a little spotting vascular lymphatics expansion, also visible massive neutrophil infiltration with lymphoid follicular hyperplasia, edema, thickening of the intestinal villi height shortened the top epithelial necrosis, loss, decrease in goblet cells, arranged in disorder; mucosal thickness, mucosal glands arranged sparse reduce glands; Chinese medicine Qingyi granules treatment group modeling 12h pathological damage has been reduced, showed reduced inflammatory cell infiltration, the villi damage shedding reduce, mucosal thickness and villus height were improved, and 24 h after modeling significant difference compared with the model group. 48 h after the indicators are close to the normal 7 groups the intestinal mucosa claudin-1 are located in the cell membrane of the intestinal epithelial tight junctions (including cell membrane at the top), small amount of staining of the membrane-proximal cytoplasmic brown staining particles nucleus and nuclear envelope no expression. The sham Claudin-1 protein expression in the cell membrane and is strongly positive; compared with the sham group, model group claudin-1 expression significantly decreased p lt; 0.05); compared with the model group, modeling of the treatment group at each time point Claudin-1 expression was elevated and statistically significant (p lt; 0.05) Conclusion: 1. applications intravenous catheter catheter via the duodenal papilla retrograde injection of 3.5% sodium taurocholate the (0.1ml/100g weight, injection speed 0.15ml / min) induced rat model of Aho prepared SAP SAP rat model is modified, SAP TCM clinical research the ideal animal research model. early application of Qingyi particles can improve SAP rat pancreatic tissue damage, alleviate the intestinal mucosa and intestinal villous atrophy, and improve the level of expression of the intestinal tight junction protein Claudin-1 level, and reduce intestinal permeability to protect the intestinal barrier and reduce mortality.
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