ABSTRACT
ObjectiveTo study the changes of serum levels of tumor n(ee)rosis factor-α (TNF-α),diamine oxidase (DAO),the expression of tight junction protein-1 (zonula occludens 1,ZO 1 ) in acute necrotizing pancreatitis (ANP) rats after ulinastatin intervention.Methods SD male rats were randomly divided into sham operation (SO) group,ANP group and ulinastatin treatment group.ANP model was induced by injecting 5％ sodium taurocholate into biliary and pancreatic duct.The rats were sacrificed at 6 and 24 hours,and then the levels of TNF-α,DAO and pathology change in pancreatic and intestinal were determined.The expression of bowel mucosa ZO-1 mRNA and protein was detected by RT-PCR and immunohistochemistry.ResultsSix hours after ANP induction,massive pancreatic necrosis and inflammatory cells infiltration were present,while epithelium necrosis of villi in ileum,vessel hemorrhage and inflammatory cells infiltration was found.The pathologic injury of pancreas and ileum in ulinastatin group was reduced when compared with that in ANP group.The serum levels of TNF-α were ( 10.83 ± 0.96),( 181.89 ± 4.93 ),( 128.23 ± 2.40) ng/L in SO group,ANP group and ulinastatin group; and the activities of DAO were (354.79 ±3.67),(117.21 ±5.58),(282.98 ± 9.12 ) U/L; the expressions of ZO 1 protein were 10.00 ± 1.87,1.20 ± 0.84,5.80 ±2.86; and the expressions ofZO 1 mRNA were 0.878 ±0.015,0.466 ±0.023,0.778 ±0.033.The serum level of TNF-α in ANP group was significantly higher than that in SO group,while the activities of DAO,expressions of ZO 1 mRNA and protein in ileum were significantly lower than that in SO group ( P ＜ 0.05 ).The serum level of TNF-α in ulinastatin group was significantly lower than that in ANP group,while the activities of DAO,expressions of ZO 1 mRNA and protein in ileum were significantly higher than that in ANP group (P ＜ 0.05).ConclusionsUlinastatin may inhibit the over release of TNF-α and improve plasma DAO activity,then increase the expression ofZO-1 mRNA and protein,thus protect the intestinal mucosa barrier.
ABSTRACT
Aim To establish the adherence of H pylori to MKN45 cell model for screening and evaluating anti-adhesive drugs.Methods H pylori suspension was incubated with FITC solution in different conditions(FITC concentration and incubation time)to optimize the labeling conditions.At the same time,Gram staining was used to determine the optimal adhesion time of H pylori to MKN45 cells.Results H pylori suspension(1?106 cuf?ml-1)incubating with FITC(5 mg?L-1)for 1 h could achieve the optimal effect for FITC-labeling H pylori.Besides,Gram staining indicated that the optimal adhesion time for H pylori to MKN45 cells was 1 h.Based on the model,the crude Aloe polysaccharide was evaluated for its anti-adhesive activity and the results showed it could significantly reduce the adherence of H pylori to MKN45 cells at the concentration of 1 g?L-1(P