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Objective To explore the expression and function of myosin light streptokinase (MLCK) in small intestine mucosa of acute necrotizing pancreatitis (ANP) rats.Methods Fifty-six male SD rats were randomly assigned to control group and ANP group.A rat model of ANP was reproduced by retrograde injection of 4％ sodium taurocholate into the biliopancreatic duct,while the control group underwent a sham operation.The rats were sacrificed at 6th,12th,24th,48th hour after ANP induction.Serum amylase、TNF α,IL 1β,diamine oxidase (DAO) were measured.The pathological scores in the pancreas and small intestine were observed.The ultrastructure and tight junction (TJ) changes in the small intestine mucosa were observed with an electron microscope.The localization and expression of MLCK in small intestine mucosa was determined by immunohistochemistry method.Results Compared to the control group,the serum amylase,TNF-α,IL-1 β,DAO level,in the ANP group were all significantly increased;[(4 978 ± 1 574) U/L vs (1 176 ± 124))U/L,(47.88 ± 15.85) μg/L vs (17.24 ± 1.99) μg/L,(132.48 ± 68.54) μg/L vs (23.51 ± 6.44) μg/L,(95.96 ± 30.84)μg/L vs (38.06 ± 17.73)U/L at 12 h],and the pathology scores of pancreas and small intestine were both significantly elevated [12 h:(12.2 ± 1.80) vs (4.68 ± 0.35),(2.58 ± 0.52) vs (0.58 ±0.26)] (P ＜0.05);the MLCK protein expression in small intestine mucosa was significantly increased in ANP group (12 h:0.1863 ± 0.0230 vs 0.1636 ± 0.0049),and the difference was statistically significant (P ＜0.05).The small intestine ultrastructure was seriously damaged and TJ was widened significantly in ANP Group.Conclusions The increased serum TNF alpha and IL-1β concentration and DAO activity and up-regulated MLCK protein expression in small intestine mucosa may damage the integrity of tight junction of intestinal epithelial cell and cause intestine mucosa barrier dysfunction.
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Objective To observe the effects of Dahuangfuzi decoction on the intestine barrier functional of acute necrotizing pancreatitis in rats. Methods The 60 rats were randomly divided into sham operation group ( n = 19 ), ANP group ( n = 21 ), and Dahuangfuzi treatment group ( n = 20). The rats of ANP group were induced by injecting 1 ml/kg of 4% sodium taurocholate into the pancreatiobiliary duct, and jejunal fistula was esablished. The rats of treatment group received Dahuangfuzi decoction (2 ml, repeated at 4 and 8 h)through jejunum distal stoma tube 0. 5 h after ANP induction. The other 2 groups received same amount of normal saline. Blood sample was collected through abdominal aorta, 24 h after ANP induction, and the serum amylase, endotoxin, D-lactate, plasma diamine oxidase (DAO) were detected. Pancreas, small intestine tissue was harvested for pathologic examination, index of intestinal epithelial damage was measured and ultrastructural changes in small intestinal mucosa was observed. Results The expression of serum amylase, endotoxin,D-lactate, DAO in sham operation group was ( 152 ± 32 ) U/L, (6.95 ± 2.10) pg/L, ( 3.96 ± 1.08 ) μg/mland ( 14.26 ± 2.67 ) μg/ml, while the corresponding values were ( 1549 ± 93 ) U/L, (40.48 ± 3.41 ) pg/L,( 12.34 ± 1.23 ) μg/ml and ( 80.28 ± 3.54) μg/ml in ANP group, and they were (655 ± 49 ) U/L, ( 19.55 ±2.50) pg/L, (6.75 ± 1.36 ) μg/mland ( 20.69 ± 7.53 ) μg/ml in treatment group. The values in ANP group were significantly higher than those in sham operation group. The values in treatment group were significantly lower than those in ANP group, but significantly higher than those in sham operation group ( P ＜ 0.05 or P ＜0. 01 ). The thickness and height of intestinal mucosa in ANP group were ( 389.44 ± 29.87 )μm and ( 16.52 ±3.73) μm, which were significantly lower than those in treatment group [(501.95 ± 45.38 )μm, (27.82 ±5.17)] μm, and in sham operation group [( 658.72 ± 57.49 ) μm, ( 35.49 ± 6.43 )μm, Index of intestional epitholial donage in ANP group was 3.72 ± 0.65 which is significently higher than those in theatment (2.12 ±0.37 ) and in sham operation group (0.85 ± 0.24). The intestinal mucosa histological and ultrastructural changes in Dahuangfuzi treatment group were better than those in ANP group. Conclusions Dahuangfuzidecoction can significantly decrease the damage of intestine barrier function in ANP rats.
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Objective: To evaluate the protective effect of N-acetylcysteine(NAC) on the small intestine of male rats after X ray irradiation of the whole abdominal region. Methods: Sixty male Spraque-Dawley rats were divided in normal control group (n=12), irradiation group(n=12), and NAC groups(50, 200, and 300 mg/kg, n=12). Irradiation injury was induced with X ray at a single dose of 1 000 cGy(source-to-skin distance 80 cm) for the abdominal regions after the animals had been anesthetized with sodium thiopental(40 mg/kg, i. p.). NAC was started 3 days before irradiation and administered for 3 more days after irradiation, and then rats were euthanized after 12 h fasting. The terminal ileum samples were collected for crypt survival assay and counting in ileal villi. The blood samples were collected for examination of superoxide dismutase(SOD) activity, and the contents of malondialdehyde(MDA) and glutathione hormone(GSH) in the small intestine. The plasma levels of D-lactate and diamine oxidase(DAO) were also measured. Results: The mucosal structure of the terminal ileum was damaged after irradiation. The plasma levels of D-lactate, DAO and the content of MDA were significantly increased; the activity of SOD and the content of GSH were significantly decreased(P<0.01). NAC at different dosages increased crypt survival rates and the number of ileal villi in the terminal ileum (P<0.05, P<0.01), enhanced the activities of SOD and content of GSH (P<0.05, P<0.01), and deceased the concentrations of D-lactate, DAO and MDA (P<0.05, P<0.01). Conclusion: NAC may protect the small intestine from irradiation-induced injury by protecting the intestinal mucosal barrier, eliminating oxygen free radicals, and maintaining the internal oxidative balance and the structure and function of intestinal mucous.
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Objective: To evaluate the protective effect of Shenfu injection (SFI) against intestinal barrier dysfunction and second hit in rats with severe acute pancreatitis (SAP). Methods: Fifty-four male Wistar rats were randomly divided into sham operation group (n = 6), SAP group (n = 24), and SAP + SFI group (10 ml/kg body wt, n = 24). Sham operation group underwent laparotomy only. SAP model was established by retrograde injection of 5% sodium taurocholate into the bilipancreatic duct of Wistar rats. SAP+SFI group was given SFI (10 ml/kg) intaperitoneally 2 h before SAP establishment. Rats were sacrificed at 3,6,12 and 24 hours after operation. The samples of pancreas, lung and liver were collected for measuring the levels of myeloperoxidase (MPO) at 3,6,12, and 24 h after injection of sodium taurocholate. The pathological changes of the terminal ilea were observed under light microscopy, and the blood levels of diamine oxidase (DAO), tumor necrosis factor alpha (TNF-α) and interleukin-6(IL-6) were also measured at different time points. Results: The blood levels of TNF-α, IL-6 and DAO in SAP group were significantly higher than those in the sham operation group at all time points (P<0.01). The levels of intrapulmonary, intrapancreatic and intrahepatic MPO in SAP group were significantly than those in the sham operation group at 6,12, and 24 hours (P<0.01). The contents of DAO, TNF-α and IL-6 were significantly decreased in SAP + SFI group compared with in SAP group at all time points (P<0.05 or P<0.01). The contents of intrapulmonary, intrapancreatic and intrahepatic MPO were significantly decreased in SAP+SFI group compared with SAP group at 6,12, and 24 h after operation (P<0.05 or P<0.01). Intestinal pathological damages were obviously milder in SAP+SFI group than that in SAP group at 24 h after operation. Conclusion: SFI can protect the small intestine mucosal barrier and other organs from second hit by reducing the polymorphonuclear leucocyte detaining and inhibiting TNF-α and IL-6.
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Objective To explore the changes in intestinal barrier function in college students with exercise-induced gastrointestinal syndrome(EIGS)for searching the related medical monitoring methods.Methods Twenty four college students underwent a six-day endurance training.Their blood D-lactic acid,lipid polysaccharide(LPS),and the activity of diamine oxidase(DAO) were tested before and after the training.Results (1) Blood D-lactic acid,LPS,and the activity of DAO increased significantly in 7 subjects with EIGS comparing to their baseline values(P<0.05):(2)The level of D-lactic acid,LPS,and the activity of DAO in the 7 subjects with EIGS were significantly higher than in those without EIGS(P<0.05).Conclusion The level of blood D-lactic acid,the content of lipid polysaccharide,and the activity of the diamine oxidase are closely related to the intestinal barrier function,and thus can be used as indicators for monitoring the stomach-intestine function during trsinjng.