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Increased Oxidative and Nitrative Stress in Human Stomach Associated with cagA+ Helicobacter pylori Infection and Inflammation

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Abstract

In order to study the role of Helicobacter pylori infection in gastric carcinogenesis, we have measured oxidized (carbonyls) and nitrated (nitrotyrosine-containing) proteins as markers for oxidative and nitrative stress in 216 human gastric biopsies using dot and western immunoblots and correlated the results with H. pylori, cagA status, expression of interleukin-8 and inducible nitric oxide synthase (iNOS) mRNAs, and gastric pathology. Higher levels of both oxidized and nitrated proteins were found in patients with either chronic gastritis or duodenal ulcer than in those with normal mucosa. The levels of modified proteins were significantly higher in inflamed samples infected with H. pylori, especially cagA+ strains, and in those with expression of interleukin-8 and iNOS mRNAs than in those negative for these parameters. These results indicate that infection with cagA+ H. pylori induces significant oxidative and nitrative stress in stomach mucosa, contributing to the pathogenesis of H. pylori-associated gastroduodenal diseases.

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REFERENCES

  1. IARC Working Group: Infection with Helicobacter pylori. In IARC Monographs on the Evaluation of Carcinogenic Risks to Humans-Schistosomes, Liver Flukes and Helicobacter pylori, Vol 61. Lyon, IARC, 1994, pp 177–240

    Google Scholar 

  2. Price AB: The Sydney system: Histological division. J Gastroenterol Hepatol 6:209–222, 1991

    Google Scholar 

  3. Drake IM, Mapstone NP, Schorah CJ, White KLM, Chalmers DM, Dixon MF, Axon ATR: Reactive oxygen species activity and lipid peroxidation in Helicobacter pylori associated gastritis: Relation to gastric mucosal ascorbic acid concentrations and effect of H. pylori eradication. Gut 42:768–771, 1998

    Google Scholar 

  4. Mannick EE, Bravo LE, Zarama G, Realpe JL, Zhang XJ, Ruiz B, Fontham ET, Mera R, Miller MJ, Correa P: Inducible nitric oxide synthase, nitrotyrosine, and apoptosis in Helicobacter pylori gastritis: Effect of antibiotics and antioxidants. Cancer Res 56:3238–3243, 1996

    Google Scholar 

  5. Baik SC, Youn HS, Chung MH, Lee WK, Cho MJ, Ko GH, Park CK, Kasai H, Rhee KH: Increased oxidative DNA damage in Helicobacter pylori-infected human gastric mucosa. Cancer Res 56:1279–1282, 1996

    Google Scholar 

  6. Farinati F, Cardin R, Degan P, Rugge M, Mario FD, Bonvicini P, Naccarato R: Oxidative DNA damage accumulation in gastric carcinogenesis. Gut 42:351–356, 1998

    Google Scholar 

  7. Fu S, Ramanujam KS, Wong A, Fantry GT, Drachenberg CB, James SP, Meltzer SJ, Wilson KT: Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis. Gastroenterology 116:1319–1329, 1999

    Google Scholar 

  8. Li CQ, Pignatelli B, Ohshima H: Coexpression of interleukin-8 and inducible nitric oxide synthase in gastric mucosa infected with cagA+ Helicobacter pylori. Dig Dis Sci 45:55–62, 2000

    Google Scholar 

  9. Pignatelli B, Bancel B, Estève J, Malaveille C, Calmels S, Correa P, Patricot LM, Laval M, Lyandrat N, Ohshima H: Inducible nitric oxide synthase, anti-oxidant enzymes and Helicobacter pylori infection in gastritis and gastric precancerous lesions in humans. Eur J Cancer Prev 7:439–447, 1998

    Google Scholar 

  10. Pignatelli B, Bancel B, Plummer M, Toyokuni S, Patricot L-M, Ohshima H: H. pylori eradication attenuates oxidative stress in human gastric mucosa. Am J Gastroenterol, In press, 2001

  11. Berlett BS, Stadtman ER: Protein oxidation in aging, disease, and oxidative stress. J Biol Chem 272:20313–20316, 1997

    Google Scholar 

  12. Shacter E, Williams JA, Lim M, Levine RL: Differential susceptibility of plasma proteins to oxidative modification: Examination by western blot immunoassay. Free Radic Biol. Med 17:429–437, 1994

    Google Scholar 

  13. Nakamura A, Goto S: Analysis of protein carbonyls with 2,4-dinitrophenyl hydrazine and its antibodies by immunoblot in two-dimensional gel electrophoresis. J Biochem Tokyo 119:768–774, 1996

    Google Scholar 

  14. Tamarit J, Cabiscol E, Ros J: Identification of the major oxidatively damaged proteins in Escherichia coli cells exposed to oxidative stress. J Biol Chem 273:3027–3032, 1998

    Google Scholar 

  15. Szabo C, O'Connor M, Salzman AL: Endogenously produced peroxynitrite induces the oxidation of mitochondrial and nuclear proteins in immunostimulated macrophages. FEBS Lett 409:147–150, 1997

    Google Scholar 

  16. Gray SF, Wyatt JI, Rathbone BJ: Simplified techniques for identifying Campylobacter pyloridis. J Clin Pathol 39:1279, 1986

    Google Scholar 

  17. Peek-RM J, Moss SF, Tham KT, Perez PG, Wang S, Miller GG, Atherton JC, Holt PR, Blaser MJ: Helicobacter pylori cagA+ strains and dissociation of gastric epithelial cell proliferation from apoptosis. J Natl Cancer Inst 89:863–868, 1997

    Google Scholar 

  18. Engstrand L, Nguyen AM, Graham DY, el Zaatari FA: Reverse transcription and polymerase chain reaction amplification of rRNA for detection of Helicobacter species. J Clin Microbiol 30:2295–2301, 1992

    Google Scholar 

  19. Beckman JS, Chen J, Ischiropoulos H, Crow JP: Oxidative chemistry of peroxynitrite. Methods Enzymol 233:229–240, 1994

    Google Scholar 

  20. Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER: Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478, 1990

    Google Scholar 

  21. Ohshima H, Celan I, Chazotte L, Pignatelli B, Mower HF: Analysis of 3-nitrotyrosine in biological fluids and protein hydrolyzates by high-performance liquid chromatography using a post-separation, on-line reduction column and electrochemical detection: Results with various nitrating agents. Nitric Oxide Biol Chem 3:132–141, 1999

    Google Scholar 

  22. Yan LJ, Orr WC, Sohal RS: Identification of oxidized proteins based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunochemical detection, isoelectric focusing, and microsequencing. Anal Biochem 263:67–71, 1998

    Google Scholar 

  23. Balabanli B, Kamisaki Y, Martin E, Murad F: Requirements for heme and thiols for the nonenzymatic modification of nitrotyrosine. Proc Natl Acad Sci USA 96:13136–13141, 1999

    Google Scholar 

  24. Poulsen HE, Prieme H, Loft S: Role of oxidative DNA damage in cancer initiation and promotion. Eur J Cancer Prev 7:9–16, 1998

    Google Scholar 

  25. Ohshima H, Bartsch H: Chronic infections and inflammatory processes as cancer risk factors: Possible role of nitric oxide in carcinogenesis. Mutat Res 305:253–264, 1994

    Google Scholar 

  26. Hahm KB, Lee KJ, Kim JH, Cho SW, Chung MH: Helicobacter pylori infection, oxidative DNA damage, gastric carcinogenesis, and reversibility by rebamipide. Dig Dis Sci 43:72S–77S, 1998

    Google Scholar 

  27. Ohshima H, Friesen M, Brouet I, Bartsch H: Nitrotyrosine as a new marker for endogenous nitrosation and nitration of proteins. Food Chem Toxicol 28:647–652, 1990

    Google Scholar 

  28. Beckman JS: Oxidative damage and tyrosine nitration from peroxynitrite. Chem Res Toxicol 9:836–844, 1996

    Google Scholar 

  29. Zingarelli B, Szabo C, Salzman AL: Reduced oxidative and nitrosative damage in murine experimental colitis in the asbsence of inducible nitric oxide synthase. Gut 45:199–209, 1999

    Google Scholar 

  30. Crabtree JE, Xiang Z, Lindley IJ, Tompkins DS, Rappuoli R, Covacci A: Induction of interleukin-8 secretion from gastric epithelial cells by a cagA negative isogenic mutant of Helicobacter pylori. J Clin Pathol 48:967–969, 1995

    Google Scholar 

  31. Tummuru MK, Sharma SA, Blaser MJ: Helicobacter pylori picB, a homologue of the Bordetella pertussis toxin secretion protein, is required for induction of IL-8 in gastric epithelial cells. Mol Microbiol 18:867–876, 1995

    Google Scholar 

  32. Liebler DC, Aust AE, Wilson GL, Copeland ES: Reactive oxidants from nitric oxide, oxidants and cellular signalling, and repair of oxidative DNA damage: A chemical pathology study section workshop. Mol Carcinog 22:209–220, 1998

    Google Scholar 

  33. Singer II, Kawka DW, Scott S, Weidner JR, Mumford RA, Riehl TE, Stenson WF: Expression of inducible nitric oxide synthase and nitrotyrosine in colonic epithelium in inflammatory bowel disease. Gastroenterology 111:871–885, 1996

    Google Scholar 

  34. Mecocci P, Fano G, Fulle S, MacGarvey U, Shinobu L, Polidori MC, Cherubini A, Vecchiet J, Senin U, Beal MF: Agedependent increases in oxidative damage to DNA, lipids, and proteins in human skeletal muscle. Free Radic Biol Med 26:303–308, 1999

    Google Scholar 

  35. Pansarasa O, Bertorelli L, Vecchiet J, Felzani G, Marzatico F: Age-dependent changes of antioxidant activities and markers of free radical damage in human skeletal muscle. Free Radic Biol Med 27:617–622, 1999

    Google Scholar 

  36. Ischiropoulos H: Biological tyrosine nitration: a pathophysiological function of nitric oxide and reactive oxygen species. Arch Biochem Biophys 356:1–11, 1998

    Google Scholar 

  37. Lamb NJ, Gutteridge JMC, Baker C, Evans TW, Quinlan GJ: Oxidative damage to proteins of bronchoalveolar lavage fluid in patients with acute respiratory distress syndrome: evidence for neutrophil-mediated hydroxylation, nitration, and chlorination. Crit Care Med 27:1738–1744, 1999

    Google Scholar 

  38. Singh VN, Gaby SK: Premalignant lesions: Role of antioxidant vitamins and beta-carotene in risk reduction and prevention of malignant transformation. Am J Clin Nutr 53:386S–390S, 1991

    Google Scholar 

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Authors
  1. Chun-Qi Li
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  2. Brigitte Pignatelli
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  3. Hiroshi Ohshima
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Li, CQ., Pignatelli, B. & Ohshima, H. Increased Oxidative and Nitrative Stress in Human Stomach Associated with cagA+ Helicobacter pylori Infection and Inflammation. Dig Dis Sci 46, 836–844 (2001). https://doi.org/10.1023/A:1010764720524

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  • DOI: https://doi.org/10.1023/A:1010764720524

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