Expression Of Cytokines And Maerix Metalloproteinases Genes In Upper Gastrointestinal Diseases Infected With Helicobacter Pylori In Children And Adolescent

Part One Expression of IL-12B,IL-10,IFN-γmRNA in gastric mucosa of upper gastrointestinal disease infected with Helicobacter pylori in children and adolescent.Objective:The aim of this study was to determine the intensity of inflammation and the expression levels of interferon-γ,(IFN-y),interleukin-12B (IL-12B),interleukin-10(IL-10)response in children and adolescent with Helicobacter pylori(H.pylori)infection.Methods:Mucosal biopsies were obtained from 132 children and adolescent,aged 3 to 18 years,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin(H&E),and gastritis was graded according to the Sydney system. Serology,PCR,Urease Test and histology were used to assess H.pylori status. Cytokines expression in gastric mucosa was confirmed in 67 patients with chronic gastritis and 30 patients with duodenal ulcer by semiquantitative reverse transcription polymerase chain reaction Biopsy-based tests. Results:In H.pylori-infected children and adolescent,most patients showed slight to moderate chronic gastritis.In gastric antrum mucosa,the levels of IFN-γand IL-12B mRNAs were significantly higher in H.pylori-infected patients than in uninfected patients.In contrast,level of IL-10 mRNA was not different between H.pylori-infected and uninfected patients.The levels of all gastric mucosal cytokine mRNAs were significantly positive correlated with the degree of antrum chronic inflammation.Conclusions:In children and adolescent,H pylori-induced inflammatory response is associated with development of cell-mediated immunity of T-helper 1 type.The studies of the natural history of H.pylori infection after initial colonization in children and adolescent are critical to understanding evolution of the host gastric mucosal inflammatory response.Such information may be critical for future treatment strategies and in particular for the development of efficacious,targeted vaccines.Part Two Expression of matrix metalloproteinase-7 and matrix metalloproteinase-9 in gastric mucosa of upper gastrointestinal disease infected with Helicobacter pylori in children and adolescent.Objective Matrix metalloproteinases(MMPs)enzymes capable of degrading extracellular matrix components,are believed to be active in tissue remodeling associated with various physiologic processes and in pathologic conditions.The aim of this study was to determine the expression of MMP-7 and MMP-9 response in children with and without H.pylori infection.Methods:Gastric biopsies were obtained from children and adolescent with dyspeptic symptoms.MMPs expression was assessed in 66 patients with chronic gastritis and 30 patients with duodenal ulcer by semiquantitative reverse transcription polymerase chain reaction Biopsy-based tests and by immuno-histochemistry using specific antibodies.Biopsy specimens were stained with hematoxilin and eosin(H&E),and gastritis was graded according to the Sydney system.Serology,PCR,Urease Test and histology were used to assess H.pylori status.Results:H.pylori induced MMP-7 expression in gastric mucosa but had a little effect on MMP-9 in children and adolescent.MMP-7 mRNA expression was higher in H.pylori infected patients than in uninfected patients.However, level of MMP-9 mRNA was not different between H.pylori-infected and uninfected patients.The levels of gastric mucosal MMP-7 mRNA was significantly positive correlated with the degree of antrum chronic inflammation. Immunohistochemical expression of MMP-7 and MMP-9 were predominantly observed at gastric epithelial cells and inflammatory cells,respectively.Conclusions:We speculate that increased expression of MMP-7 production is an early event in the response to gastric Helicobacter infection,a feature that may favor the tissue remodeling and recruitment of immune cells early during infection.An enhanced understanding of the molecular mechanisms responsible for the activation of MMP-7 may lead to a new therapeutic strategy. Objective The purpose of this study was to investigate the relation between the promoter of IL-12B,MMP-7,MMP-9 gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without H.pylori infection in children and adolescent.Methods:Genomic DNA was obtained from peripheral blood or gastric biopsies of 100 patients with chronic gastritis and 32 patients with duodenal ulcer and 102 healthy subjects.The promoter of IL-12B+1188A/G, MMP-7-181A/G,MMP-9-1562 C/T gene polymorphisms were genotyped by polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP)and sequencing.The genotype distributions and allele frequencies were compared between patients and controls,and the association of genotypes with clinicopathological features was studied.Results:The genotype distributions and allele frequencies of IL-12B+ 1188A/G,MMP-7-181A/G,MMP-9-1562 C/T gene polymorphisms were similar in gastric upper gastrointestinal disease and healthy subjects.The C/C genotype genotype of MMP-9-1562C/T gene polymorphism was associated with Helicobacter pylori status.IL-12B,MMP-7 and MMP-9 gene polymorphisms did not affect their mRNA level expression and were not associated with the degree of antrum chronic inflammation.Conclusions:These data suggest that IL-12B,MMP-7 and MMP-9 gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.However,the C/C genotype of MMP-9-1562 C/T gene polymorphism might be associated with H.pylori infection.