1. The role of the IL 6 190 C/T and IL/6 174 C/G polymorphisms in the development of gastritis in children Authors: Mărginean Maria Oana, Cosmin Oprea, Ioan Serban Fanfaret, Cristian Dan Mărginean Coordinators: Professor Bănescu Claudia, Assistant professor Pitea Ana Maria

2. Introduction Polymorphisms in the promoter region of the proinflammatory cytokine, interleukin (IL)-6 have been related to several chronic inflammatory diseases, including children gastritis Genetic variability: –of the gene IL-6-190 - allele T or C –of the gene IL/6 174 – allele C or G Asociated with various hereditary diseases, including gastritis

3. Introduction Gastritis inflammatory process in the gastric mucosa, with histopathological modifications commonly associated with peptic ulcer and duodenitis great importance and certain actuality in the field of pediatric pathology because of its increased incidence in children and its complications

4. Background Recent data suggests that gastritis may have an inflammatory etiology associates a low degree proinflammatory status Some cases of gastritis associated with H. pylori infection do not heal despite the correct application of treatment regimens due to the virulence of microorganisms in association with the hosts` genetic predisposition

5. Pathogenesis imbalance between aggressive factors and the defense mechanisms aggressive factors –acid secretion, pepsin secretion, stress, aspirin, NSAIDs, H. pylori defense mechanisms –mucus and bicarbonate secretion –mucosal blood flow –prostaglandines (PG)- stimulate other mechanisms

6. Pathogenesis The etiology of gastroduodenal diseases –environmental factors –genetic predisposition consisting of genetic susceptibility to infection –DNA repair mechanisms related to carcinogenesis host polymorphism –influence the pharmacokinetics –efficacy of treatment with proton pump inhibitors

7. Background Genetic polymorphisms of several inflammatory and immunoregulatory cytokines are investigated for possible association with risk for specific H. pylori- associated disease Il-6 is a multifunctional cytokine produced by immune or nonimmune cells, and functions as an inflammatory endocrine and metabolic function mediator There is a correlation between H. pylori and IL-6, as mRNA levels of this cytokine in the gastric mucosa have been correlated with the level of inflammation. Normally, IL-6 plays an important role in host defense mechanisms as a messenger between innate and adaptative systems by stimulating interferon gamma production in T-cells, by promoting immunoglobulin secretion in activated B-cells, and through polymorphoneutrophil activation

8. Objective to establish the role of IL 6 in children gastritis to establish correlations between the polymorphisms of the gene IL 6 190 C/T, IL 6 174 C/G and children gastritis to determine whether patients differ from children without digestive pathology regarding genotype polymorphisms distribution.

9. Material and method 123 children admited in the Pediatrics Clinic I –from january 2014 through march 2015 with symptoms such as: »abdominal pain »nausea, vomiting »Pirozis -we correlated these symptoms with endoscopical changes, histopatological changes and Helicobacter Pylori infection The patients were divided according to their histopathological changes into: a control group - 55 children without histopathological changes a study group – 68 children with histopathological changes The following were evaluated: - clinical symptoms - IL-6 190 T/C, IL 6 174 C/G polymorphism analysis (using specific primers)

10. Material and method Inclusion criteria: –Children with symptoms which were characteristic for gastritis (pirosis, nausea, vomiting) Control group – children without histopathological changes in the antral region Study group - children with histopathological changes of the gastric mucosa Exclusion criteria: –children with infectious diseases –children with an acuta abdomen –children without any symptoms than those which were already mentioned

11. Material and method Statistical calculation –for the entire statistical calculation Graph Pad 3.6 Software, San Diego, California, USA –T Student test was used to assess the differences between the means of continuous variables (expressed as mean ± SD) –The differences among constant variables and three genotype groups of the IL 6 190 C/T, IL/6 174 C/G and polymorphisms were estimated using ANOVA and Kruskal Wallis tests, an analysis which is appropriate for more than two groups.

12. Results Average age of the children included in the study: –9.67 ± 3.52 years in the control group –9.21± 2.11 years in the study group without a statistically significant diference between the two groups

13. Results The distribution of the cases according to the IL 6 190 C/T genotypes was the following: LotsIL 6 190Total CCCTTT Control group 49,1%38,2%12,7%100,0% Study group 35,3%44,1%20,6%100,0% We observed that the genotype CT was more frequent in the study group, while the genotype CC was more frequent in the control group, but with a statistically insignificant p value( p = 0.47/0.16).

14. Results- polymorphism IL 6 190 C/T No correlation between this polimorphism, clinical symptoms and histopatological changes was observed. p = 0.20

15. Results- polymorphism IL 6 190 T/C P = 0.34 P = 0.15

16. Results- polymorphism IL 6 190 C/T HPylori+HPylori-ORIC (95%)P VALUE TT REF219--- CT10412.310.46-11.60.48 CC7441.510.8-7.960.95 TC+CC17851.90.40-8.930.52 Allela T1479--- Allela C241291.050.51-2.50.89 The IL 6 190 C/T polimorphism does not correlate with the risk of developing an H. Pylori infection.

17. Results- polymorphism IL 6 174 C/G Lots IL 6 174TotalP CCCGGG Control group 21,8%34,5%43,6%100,0% 0.71 Study group 7,4%77,9%14,7%100,0% 0.16 The distribution of the cases according to the IL 6 174 C/G genotypes was the following: A higher frequence of the CG genotype of the IL 6 174 polymorphism was observed in the study group (p<0.05), while the genotype GG was more frequent in the control group (p < 0.05).

18. Results- polymorphism IL 6 174 C/G P = 0.03/0.05 Vomiting is correlated with the genotype CG and GG of the IL 6 174 C/G gene in the study group (with histopathological changes)

19. Rezultate polimorfism IL 6 174 C/G Loss of appetite is correlated with the genotypes CG and GG of the IL 6 174 C/G gene in the study group P = 0.001

20. There is a risk of developing an H Pylori infection for those who have the CG and GG polymorphism ( p < 0.05) HPylori+HPylori-ORIC (95%)P VALUE GG REF529--- CG14581.41.45-4.20.05 CC0170.150.007-2.80.15 GC+CC14751.080.5-3.250.94 Allela G24116--- Allela C14921.730.36-2.50.05 Results- polimorphism IL 6 174 C/G

21. Results- polymorphism IL 6 174 C/G Alelle C of the IL 6 174 C/G polymorphism is statistically significant correlated with H Pylori infection [OR of 1.73, 95% CI (0.36-2,50)] ( p = 0.05).

22. Conclusions IL 6 174 is an important parameter of inflammation, correlated with inflamation and child gastritis. Gastritis is more frequently found in the children which have the CG genotype of the IL 6 174 gene polymorphism Vomiting, appetite loss and histopathological changes were correlated especially with genotypes CG and GG of the IL 6 174 gene. The disease was not correlated with the IL 6 190 gene polymorphism. Further studies are warranted to investigate the relation between IL 6 polymorphism, H. pylori infection and response to terapy.