Association of H pylori cagA and vacA genotypes and IL-8 gene polymorphisms with clinical outcome of infection in Iranian patients with gastrointestinal diseases

AIM： To find out if a functional promoter polymorphism in the IL-8 gene along with cagA status and polymorphisms in vacA gene influence the type of diseases in Iranian patients infected by Hpylori. METHODS： IL-8 -251 A/T polymorphism was genotyped by oligonucleotide allele specific PCR （ASO-PCR） in a sample of 233 patients with Hpylori infection undergoing upper gastrointestinal endoscopy. The presence of cagA gene and polymorphisms in vacA gene was also determined by PCR. Association of these genetic polymorphisms with the development of gastritis, peptic ulcers as well as gastric cancer was tested. RESULTS： When the patients with different clinical manifestations were compared according to the presence of cagA gene or various vacA genotypes, only the vacA genotypes were significantly different among gastritis, peptic ulcer and gastric cancer patients （χ^2= 17.8; P=0.001）. Furthermore, there was a significant difference in the frequency of IL-8 -251 A/T genotypes between patients with gastric cancer and benign diseases （χ^2=10.47;P=0.005） CONCLUSION： The IL-8 -251 A/T polymorphism and the polymorphisms in H pylor/ vacA gene are involved in limiting the infection outcome to gastritis and peptic ulcer or in favoring cancer onset in Iranian patients.

Evaluation of interleukin 8 +2767 A/T polymorphism in visceral leishmaniasis

Objective:To evaluated the relationship between the genetic variations at IL-8 +2767 position with VL pathogenesis among Iranian patients.Methods:Three groups including patients with VL clinical presentation and leishmania seropositive(n=124).patients seropositive but without clinical presentation(n=82) and healthy controls(n=63) were selected to conduct this cross-sectional study.Polymorphism at +2767 position of IL-8 was investigated using PCR-RPLP techniques.Anti-leishmania antibody titration was evaluated by the immunoflorescence technique.Results:We observed higher significant frequencies +2767 A/A and A/T genotypes in groups I compared to group 2 and healthy controls(P=0.001).Also,patients in Group 1 carriyng A/A genotype showed higher liter of antileshmania antibody than patients with A/T and T/T genotypes(P=0.05).The validity of the data was analyzed using Hardy-Weinberg equilibrium and one way analysis of variance(ANOVA),as well as x^2tests.Conclusions:Our findings indicate that the IL-8 +2767 polymorphism is significantly involved in impaired immune responses against VL and it could be considered as a risk factor for the VL progress.

Expression of CXC receptor 1 and 2 in esophageal mucosa of patients with reflux esophagitis

AIM: Interleukin 8 (IL-8) mediates neutrophil trafficking via its receptors. Recent studies have shown that IL-8 is likely involved in the development and progression of erosive reflux esophagitis (RE), yet little is known about the two distinct receptors, CXC receptor (CXCR)-1 and -2.The purpose of this study was to determine CXCR-1 and -2 messenger RNA expression levels in RE.METHODS: We studied 26 patients with RE and 15asymptomatic controls. Paired biopsy samples were taken from the esophagus 3 cm above the gastroesophageal junction; one biopsy was snap frozen for measurement of CXCR-1 and -2 mRNA levels by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and another was formalin-fixed for histopathological evaluation.We also examined the association of the expression levels of CXCR-1 and -2 mRNA with histopathological hallmarks of RE.RESULTS: The relative CXCR-1 and -2 mRNA expression levels were rather decreased in esophageal mucosa of patients with RE, compared to those in normal esophagus of controls. There were no significant difference in the relative mRNA expression levels of CXCR-1 and -2 among endoscopic grades of RE based on the Los Angeles classification. Each histopathological hallmark of GERD was not associated with the expression levels of CXCR-1 and -2 mRNA.CONCLUSION: Apart from overexpression of IL-8, the relative expression levels of CXCR-1 and -2 mRNA were rather lower than expected in the affected esophageal mucosa of patients with RE.

Expression and cellular localization of interleukin 8 mRNA and protein in the area of xenogenic bone implant

Objective: To observe the expression and cellular localization of interleukin 8 (IL 8) mRNA and protein in the area of xenogenic bone implant. Methods: The bovine cancellous bone granules were implanted into the thigh muscles of mice. The samples were taken 4, 7, 14 and 21 days after implantation. IL 8mRNA and protein in the site of implant were assayed by in situ hybridization and immunocytochemical techniques. Results: The expression of IL 8mRNA and protein were observed in all specimens 4, 7, 14 and 21 days after implantation. IL 8mRNA was expressed mainly by the neutrophils, monocytes, macrophages and fibroblasts at 7th day post implantation. Some mesenchymal cells, multinucleated giant cells, vascular endothelial cells and smooth muscle cells also expressed IL 8mRNA in the area of xenogenic bone implant at 14th and 21st days. Immunocytochemical studies demonstrated the same results as that of in situ hybridization. Conclusions: Many different kinds of cells express IL 8mRNA and secret IL 8 in the area of xenogenic bone implant, suggesting that IL 8 may play an important role in local immunity of xenogenic bone graft.

Effect of cigarette smoke extraction on the expression of found in inflammatory zone 1 in rat lung epithelial L2 cells

Background Found in inflammatory zone 1 （FIZZ1） protein increased in pulmonary epithelial cells and in limited amounts of other lung cells.FIZZ1 increased in murine model of smoke induced chronic obstructive pulmonary disease.However,the direct role of FIZZ1 produced by pulmonary epithelium stimulated with cigarette smoke extraction has not been determined.We examined the expression and function of FIZZ1 in rat lung epithelial L2 cells.Methods The rat lung epithelial L2 cells （CCL 149） were exposed to cigarette smoke extraction,expression of FIZZ1 mRNA was investigated by RT-PCR.Levels of FIZZ1 protein were detected by Western blotting and laser confocal microscope.CCL 149 cells were treated with different concentrations and for different time of recombinant protein FIZZ1.After treatment,the expression levels of interleukin 8 （IL-8） were detected by enzyme-linked immunosorbent assay （ELISA）.Results When CCL 149 cells were exposed to cigarette smoke extraction,FIZZ1 mRNA and protein levels expressed significantly higher than control group.Recombinant protein FIZZ1 promoted the expression of IL-8 in a dose and time dependent manner in a certain range.Conclusions Cigarette smoke extraction activates FIZZ1 at mRNA and protein levels in CCL 149 cells.Recombinant protein FIZZ1 induces the expression of IL-8 and may thus participate in the process of chronic obstructive pulmonary disease airway inflammation and airflow obstruction.Generally,immune cells such as macrophages,neutrophils and lymphocytes are unavoidably involved in airway inflammatory and immune responses to cigarette smoke,but it is still unclear whether their involvement in the pathogenesis of chronic obstructive pulmonary disease is based on the specific expression in lung epithelial cells of FIZZ1.

Anti-helicobacter pylori effect of total alkaloids of sophora alopecuroides in vivo

Background Helicobacterpylori （H.pylori） infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer.Total alkaloids of sophora alopecuroides （TASA） is widely used in herbal remedies to treat various infectious diseases,including stomach-associated diseases.This study is aimed at evaluating the antimicrobial activity of TASA on H.pylori-infected BALB/c mice mouse gastritis.Methods Totally 120 BALB/c mice were orally inoculated with H.pylori Bacterial liquid to construct BALB/c mice H.pylori infection gastritis animal model,after the model was successfully created.We randomly assigned 100 infected mice into 10 treatment groups,the first group （normal saline）; the second group （bismuth pectin）; the third group （omeprazole）; the fourth group （TASA 2 mg/d）; the fifth group （TASA 4 mg/d）; the sixth group （TASA 5 mg/d）; the seventh group （TASA ＋ bismuth pectin）; the eighth group （TASA ＋ omeprazole）; the ninth group （bismuth pectin ＋ clarithromycin ＋ metronidazole）;the tenth group （omeprazole ＋ clarithromycin ＋ metronidazole）,5 other non-infected mice as negative control.Mice were orally inoculated twice a day and 7 days continuously.Then the mice were killed 4 weeks after treatment,we used realtime PCR to detect 16sDNA of H.pylori to test both the colonization and the clearance mice of bacteria of each treatment.We applied hematoxylin and eosin （HE） staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 （IL-8）,cyclooxygenase 2 （COX-2）,and nuclear factor-kappa B （NF-KB） expression change after treatments.Results Firstly,we ensured that after 6-week intragastric administration,the bacteria colonization reached an exceed peak which is far higher than positive threshold （P ＜0.001）; secondly,after treatments,it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H.pylori as well as conventional triple therapy （P ＜0.001）; thirdly,HE staining showed that the inflammation on mice gastric mucosal membrane were also relieved obviously in TASA combined treatments and conventional triple therapy compared with normal saline treated mice,moreover,from immunohistochemistry results,H.pylori-induced IL-8,COX-2,and NF-KB were consistently suppressed in seventh,eighth,ninth,and tenth group to a certain extent.Conclusion These results open the possibility of taking TASA as an anti-inflammatory agent for H.pylori gastritis.