IL-6 and IL-8 in H. pylori-associated Gastritis (Siregar GA, et al. Indones Biomed J. : 163-6 DOI: 10. 18585/inabj. RESEARCH ARTICLE Comparison of IL-6. IL-8 Concentrations in pylori- and non-H. pylori-associated Gastritis Gontar Alamsyah Siregar1,E. Sahat Halim2. Ricky Rivalino Sitepu2 Division of Gastroentero-Hepatology. Department of Internal Medicine. Faculty of Medicine. University of Sumatera Utara/ Adam Malik General Hospital. Jl. Bunga Lau No. Medan. Indonesia Department of Internal Medicine. Faculty of Medicine. University of Sumatera Utara/ Adam Malik General Hospital. Jl. Bunga Lau No. Medan. Indonesia Corresponding author. E-mail: gontarsiregar@gmail. Abstract Abstrak ACKGROUND: Helicobacter pylori is a noninvasive microorganism causing intense gastric mucosal inlammatory and immune reaction. The gastric mucosal levels of the proinlammatory cytokines Interleukin 6 (IL-. and IL-8 have been reported to be increased in H. pylori infection, but the serum levels in H. pylori infection is still controversial. The purpose of this study was to investigate the serum levels of IL-6 and IL-8 in pylori infection. ATAR BELAKANG: Helicobacter pylori merupakan mikroorganisme non invasif yang menyebabkan inlamasi mukosa gaster dan reaksi Kadar sitokin proinlamasi Interleukin 6 (IL-. dan IL-8 di mukosa gaster telah dilaporkan meningkat pada pasien yang terinfeksi H. Sementara kadar sitokin proinlamasi di serum masih kontroversi. Tujuan penelitian ini untuk mengetahui kadar serum IL-6 dan IL-8 pada gastritis yang berkaitan dengan infeksi H. METHODS: A cross sectional study was done on eighty consecutive gastritis patients admitted to endoscopy units at Adam Malik General Hospital and Permata Bunda Hospital. Medan. Indonesia from May-October 2014. Histopathology was performed for the diagnosis of gastritis. Rapid urease test for diagnosis of H. pylori infection. Serum samples were obtained to determine circulating IL-6 and IL-8. Univariate and bivariate analysis . ndependent t tes. were done. METODE: Studi potong lintang secara prospektif terhadap 80 pasien gastritis secara konsekutif yang datang ke unit endoskopi Rumah Sakit Umum Adam Malik dan Rumah Sakit Permata Bunda. Medan. Indonesia dari Mei-Oktober Diagnosis gastritis dilakukan secara histopatologi. Rapid urease test digunakan untuk diagnosis infeksi H. Dilakukan pemeriksaan kadar serum IL-6 dan IL8. Dilakukan analisis univariat dan bivariat . ndependent t RESULTS: There were 41. 25% patients infected with H. Circulatory IL-6 levels were signiicantly higher in pylori-infected patients compared to H. pylori negative, but there were no differences between serum levels of IL-8 in H. pylori positive and negative patients. CONCLUSION: The immune response to H. promotes systemic inlammation, which was relected in an increased level of serum IL-6. Serum levels of IL-8 were not signiicantly different between H. pylori positive and HASIL: Terdapat 41,25% pasien terinfeksi H. Kadar serum IL-6 lebih tinggi secara signiikan pada pasien yang terinfeksi H. pylori dibandingkan H. pylori negatif, sementara tidak ada perbedaan signiikan antara kadar serum IL-8 pada pasien dengan H. pylori positif dan negatif. KESIMPULAN: Respons imun terhadap H. menyebabkan inlamasi sistemik, seperti terlihat pada peningkatan kadar serum IL-6 secara signiikan. Kadar serum IL-8 tidak berbeda secara signiikan pada kedua The Indonesian Biomedical Journal. Vol. No. December 2014, p. KEYWORDS: Helicobacter pylori, gastritis. IL-6. IL-8. Print ISSN: 2085-3297. Online ISSN: 2355-9179 KATA KUNCI: Helicobacter pylori, gastritis. IL-6. IL-8. Indones Biomed J. : 163-6 Introduction Helicobacter pylori is the most common bacterial infection of humans that is speciic for gastric epithelial cells. Depending on geographic location. pylori is inhabitant of half of the world populationAos stomach. pylori plays a crucial role in the development of gastrointestinal diseases such as chronic gastritis, gastric ulcer, and duodenal ulcer mucosa associated lymphoid tissue lymphoma and gastric pylori is a non-invasive microorganism causing intense gastric mucosal inlammatory and immune . Cytokines are responsible for orchestrating the immune response. pylori-induced gastric mucosal cytokine overproduction has been clearly documented Local production of proinlammatory cytokines such as interleukin 6 (IL-. and IL-8 are thought to play a central role in the recruitment of inlammatory cells to the gastric mucosa in the presence of H. Whether this inlammation is limited to gastric mucosa or causes systemic inlammation remains unclear. The stomach has a large surface area and continuous spillover of locally produced cytokines into the blood stream is a There are few and conlicting data on circulatory cytokine levels in patients with H. pylori infection. The purpose of the present study was to compare the serum levels of IL-6 and IL-8 as proinlammatory cytokines in H. pylori- and non-H. pylori-associated gastritis patients. Methods Patient selection The present study was a cross sectional study on gastritis patients at Adam Malik General Hospital and Permata Bunda Hospital. Medan. Indonesia from May-October 2014. Eighty consecutive patients with gastritis admitted to endoscopy units were included in the study. All patients gave informed consent and the study was approved by the local ethic committee. No. 331/KOMET/FK USU/2014. None of the patients had received antibiotics, bismuth compounds. H2 antagonists, proton pump inhibitors or immune modulating drugs within the last four weeks before endoscopy. Patients with evidence of malignancy, immunosuppression, metabolic disorders, or gastrointestinal hemorrhage and patients who had a history of gastric surgery were excluded. Diagnosis of gastritis Diagnosis of gastritis was evaluated from biopsy of the mucosa of gastric antrum and body. The biopsy specimens were ixed in 10% formalin and embedded in parafin. The samples were stained using Hematoxylin-Eosin and were evaluated by an anatomic pathologist from Faculty of Medicine. University of Sumatera Utara. Diagnosis of H. pylori was considered positive from the positive results of rapid urease test. Result of the rapid urease test was read within 24 hours. Serum levels of IL-6 and IL-8 Venous blood was drawn using a serum separator tube and allowed to clot for 30-45 minutes at room temperature before centrifugation for 15 minutes at approximately 1,000g. Serum was immediately stored frozen in aliquots at -20AC until assays for IL-6 and IL-8 were performed. IL-6 was assayed with Quantikine ELISA Human IL-6 Immunoassay (R&D System. Minneapolis. MN. USA), while IL-8 was assayed with Quantikine HS ELISA Human CXCL8/IL-8 Immunoassay (R&D Syste. Statistical analysis SPSS version 22 (SPSS Inc. Chicag. was used for the The data were analyzed using univariate and bivariate analysis with 95% conidence interval. Results are expressed as the mean /standard deviation. Bivariate analysis was carried out using independent t test. p60 5%) 18 . Job Self employed 11 . 75%) 15 . 75%) 26 . Employee 5 . Farmer 3 . Housewife 9 . Other 5 . Nutritional status Normal Under- & Over-weight 17 . Total 27 . 75%) 43 . 25%) 47 . 75%) 44 . Figure 1. Mean Levels of IL-6 and IL-8 in H. pylori ( ) and pylori (-) Patients. Table 2. Serum level of IL-6 and IL-8 in H. pylori ( ) and pylori (-) Serum Level H. pylori positive H. pylori negative Independent . ean SD) t test . g/m. ean SD) IL-6 06 9. IL-8 *p<0. Discussion Mean age of gastritis subjects in this study was 49. (SD) years old, which is considered a productive age group. In addition, the age groups with the most cases of gastritis were those in the age group of 46-60 and 30-45. This result is in accordance with the results from the previous studies such as that by Garg B, et al. , which reported the mean age of gastritis patients was 47 years and the study by Mustapha SK, et al. , which reported a mean age of 47. 2 years. The majority of previous studies on cytokines have focused on patients with severe gastritis or peptic ulcer. this study, a group of patients with gastritis without major gastric diseases, such as peptic ulcer and gastric cancer, was examined with the purpose to describe gastritis in patients with or without H. pylori infections. Both clinical trial and population based epidemiologic studies achieved conlicting results about blood levels of inlammation markers in H. pylori infection. This study concern about serum levels of IL-6 and IL-8 as proinlammatory cytokine in H. pyloriinfected patients without systemic diseases. pylori is the major recognized etiological agent inducing gastric inlammatory responses. These responses can be considered to have two components: chronic inlammatory responses associated with increased numbers of mononuclear cells in the lamina propria iniltration, including lymphocytes, monocytes/macrophages, and plasma cells. and acute inlammatory response characterized by intraepithelial and interstitial neutrophilic. These two responses may be regulated differentially following induction of cytokines involved in the inlammatory cascade. IL-6 may be involved in chronic inlammatory changes through its broad effects on growth and differentiation of mononuclear cells, including T and B lymphocytes and macrophages. IL-6 correlated strongly with the mononuclear cell density. ,7,. Yamaoka et al. showed that mucosal IL-6 levels were higher in early gastric cancer with active H. pylori infection than without H. pylori infection. Lindholm et al. also detect increased levels of mucosal IL-6 in the H. pyloriinfected subjects. This study found that circulating level The Indonesian Biomedical Journal. Vol. No. December 2014, p. of IL-6 was signiicantly higher in infected group compared to H. pylori negative. Other study reported the same result. Nakagawa et al. reported that higher H. pylori IgG levels were signiicantly associated with higher serum IL-6 levels among H. pylori-infected individuals. Several investigators have showed that IL-8 is an important mediator in H. pylori-associated gastritis. contrast to IL-6. IL-8 is speciically implicated in the pathogenesis of infectious and inlammatory conditions associated with neutrophil iniltration because of its potent chemotactic and stimulatory activity for neutrophils. IL-8 is the most likely substance responsible for inducing further steps of the signal transducing pathway because it is upregulated in epithelial cells due to H. pylori infection. Lindholm et al. found increased levels of mucosal IL-8 in the H. pylori-infected subjects. Fan et al. showed that the concentrations of IL-8 in gastric juice and gastric biopsy homogenates were substantially greater in patients infected with H. pylori, but there were no signiicant differences of IL-8 concentrations between H. pylori positive and negative . Cichoz-Lach et al. found that mean level of IL-8 in patients with biliary gastritis was higher than the control group, and was more increased in H. pylori infected patients than in uninfected ones. In erosive gastritis accompanied by H. pylori infection mean level of IL-8 was higher than in uninfected patients for which the level was normal. This study also found that serum levels of IL-8 were higher in H. pylori-associated gastritis, but not signiicant compared to H. pylori negative. Kim et al. showed that pylori strains that express cytotoxin-associated gene A (CagA) were found to upregulate epithelial IL-8 secretion and gene expression, a direct CagA effect on IL-8 induction by gastric epithelial cells. We did not determine the CagA status of our patients. This can be our limitation for this study, so we possibly had patients infected with less pathogenic strains of this bacterium such as with CagAnegative H. This might be a reason that there were no signiicant differences between serum levels of IL-8 in pylori positive and negative. Our indings suggest that a strong immune response to H. pylori enhanced the systemic inlammation, which was relected in an increased level of serum IL-6. Print ISSN: 2085-3297. Online ISSN: 2355-9179 References