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Molecular and Cellular Biochemistry

, Volume 270, Issue 1–2, pp 125–130 | Cite as

Antimicrobial effects of antioxidants with and without clarithromycin on Helicobacter pylori

  • Archana Chatterjee
  • Debasis Bagchi
  • Taharat Yasmin
  • Sidney J. Stohs
Article

Abstract

Increasing resistance to currently used antimicrobials has resulted in the evaluation of other agents that have antimicrobial activity against Helicobacter pylori. H. pylori American Type Culture Collection (ATCC) strain 49503 (a toxin-producing strain known to be associated with gastric cancer) was grown, a cell suspension prepared in 2 mL PBS and diluted 10-fold. One hundred μL of this cell suspension was added to vitamin C 0.5%, vitamin E 0.5%, garcinol 100 μg/mL, Protykin® (containing 50% rans-resveratrol) 100 μg/mL and garcinol + Protykin® 100 μg/mL in Lennox broth, and incubated for 16 h under microaerophilic conditions. Three replicates of 10 μL from each 10−7 dilution tube were plated, colonies were counted after 16 h, and growth of H. pylori was confirmed by the CLO® test. These colony counts were compared to control cultures without the addition of any antioxidants. The experiments were then repeated with the addition of 15 μg/mL of clarithromycin to experimental and control samples. Enhanced killing of H. pylori by 37.6% was noted when vitamin C was added, which increased to 66% when clarithromycin was added, compared to controls (p < 0.05). With garcinol and Protykin® alone there was 91.4 and 87% killing of H. pylori, respectively, while a combination of garcinol + Protykin® resulted in 90.8% killing compared to controls (p < 0.05). When clarithromycin was added, there was 76.3% increased killing with garcinol alone, 55.3% with Protykin® alone, and 73.7% with garcinol + Protykin® compared to controls (containing clarithromycin) (p < 0.05). Vitamin E had no effect on H. pylori growth compared to controls. We conclude from this study that some antioxidants such as vitamin C, garcinol and Protykin®, but not vitamin E, may have potential as antimicrobial agents against H. pylori. (Mol Cell Biochem 270: 125–130, 2005)

Key words

antioxidants clarithromycin Helicobacter pylori 

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References

  1. 1.
    Wang J, Brooks EG, Bamford KB, Denning TL, Pappo J, Ernst PB: Negative selection of T cells by Helicobacter pylori as a model for bacterial strain selection by immune evasion. J Immunol 167: 926–934, 2001PubMedGoogle Scholar
  2. 2.
    Yang YJ, Yang JC, Jeng YM, Chang MH, Ni YH: Prevalence and rapid identification of clarithromycin-resistant Helicobacter pylori isolates in children. Pediatr Infect Dis J 20: 662–666, 2001Google Scholar
  3. 3.
    Moss SF: The carcinogenic effect of H. pylori on the gastric epithelial cell. J Physiol Pharmacol 50: 847–856, 1999Google Scholar
  4. 4.
    Duck WM, Sobel J, Pruckler JM, Song Q, Swerdlow D, Friedman C, Sulka A, Swaminathan B, Taylor T, Hoekstra M, Griffin P, Smoot D, Peek R, Metz DC, Bloom PB, Goldschmidt S, Parsonnet J, Triadafilopoulos G, Perez-Perez GI, Vakil N, Ernst P, Czinn S, Dunne D, Gold BD: Antimicrobial resistance incidence and risk factors among Helicobacter pylori-infected persons, United States. Emerg Infect Dis 10: 1088–1094, 2004Google Scholar
  5. 5.
    Thyagarajan SP, Ray P, Das BK, Ayyaragi A, Khan AA, Dharmalingam S, Rao UA, Rajasambandam P, Ramathilagam B, Bhasin D, Sharma MP, Naik SR, Habibullah CM: Geographical differences in antimicrobial resistance pattern of Helicobacter pylori clinical isolates from Indian patients: Multicentric study. J Gastroenterol Hepatol 18: 1373–1378, 2003Google Scholar
  6. 6.
    Passaro DJ, Chosy EJ, Parsonnet J: Helicobacter pylori: Consensus and controversy. Clin Infect Dis 35: 298–304, 2002CrossRefGoogle Scholar
  7. 7.
    Gold BD: Helicobacter pylori infections: Epidemiology and current issues in diagnosis and management. Pediatric News (Suppl): 3–11, 2003Google Scholar
  8. 8.
    Toracchio S, Marzio L: Primary and secondary antibiotic resistance of Helicobacter pylori strains isolated in central Italy during the years 1998–2002. Dig Liver Dis 35: 541–545, 2003Google Scholar
  9. 9.
    Chatterjee A, Yasmin T, Bagchi D, Stohs SJ: The bactericidal effects of Lactobacillus acidophilus, garcinol and Protykin® compared to clarithromycin, on Helicobacter pylori. Mol Cell Biochem 243: 29–35, 2003Google Scholar
  10. 10.
    Sharon N, Ofek I: Fighting infectious diseases with inhibitors of microbial adhesion to host tissues. Crit Rev Food Sci Nutr 42: 267–272, 2002Google Scholar
  11. 11.
    Burger O, Weiss E, Sharon N, Tabak M, Neeman I, Ofek I: Inhibition of Helicobacter pylori adhesion to human gastric mucus by a high-molecular-weight constituent of cranberry juice. Crit Rev Food Sci Nutr 42: 279–284, 2002PubMedGoogle Scholar
  12. 12.
    Burger O, Ofek I, Tabak M, Weiss EI, Sharon N, Neeman I: A high molecular mass constituent of cranberry juice inhibits Helicobacter pylori adhesion to human gastric mucus. FEMS Immunol Med Microbiol 29: 295–301, 2000Google Scholar
  13. 13.
    Nariman F, Eftekhar F, Habibi Z, Falsafi T: Anti-Helicobacter pylori activities of six Iranian plants. Helicobacter 9: 146–151, 2004Google Scholar
  14. 14.
    Bagchi D, Bhattacharya G, Stohs SJ: Production of reactive oxygen species by gastric cells in association with Helicobacter pylori. Free Radic Res 24: 439–450, 1996Google Scholar
  15. 15.
    Bagchi D, McGinn TR, Ye X, Bagchi M, Krohn RL, Chatterjee A, Stohs SJ: Helicobacter pylori-induced oxidative stress and DNA damage in a primary culture of human gastric mucosal cells. Dig Dis Sci 47: 1405–1412, 2002Google Scholar
  16. 16.
    Park JH, Kim SY, Kim DW, Lee WG, Rhee KH, Youn HS: Correlation between Helicobacter pylori infection and vitamin C levels in whole blood, plasma and gastric juice, and the pH of gastric juice in Korean children. J Pediatr Gastroenterol Nutr 37: 53–62, 2003Google Scholar
  17. 17.
    Sasazuki S, Sasaki S, Tsubono Y, Okubo S, Hayashi M, Kakizoe T, Tsugane S: The effect of 5-year vitamin C supplementation on serum pepsinogen level and Helicobacter pylori infection. Cancer Sci 94: 378–382, 2003Google Scholar
  18. 18.
    Sjunnesson H, Sturegard E, Willen R, Wadstrom T: High intake of selenium, beta-carotene, and vitamins A, C, and E reduces growth of Helicobacter pylori in the guinea pig. Comp Med 51: 418–423, 2001Google Scholar
  19. 19.
    Sanderson MJ, White KL, Drake IM, Schorah CJ: Vitamin E and carotenoids in gastric biopsies: the relation to plasma concentrations in patients with and without Helicobacter pylori gastritis. Am J Clin Nutr 65:101–106, 1997Google Scholar
  20. 20.
    Yamaguchi F, Ariga T, Yoshimura Y, Nakazawa H: Anti-oxidative and anti-glycation activity of garcinol from Garcinia indica fruit rind. J Agric Food Chem 48: 180–185, 2000Google Scholar
  21. 21.
    Mahady GB, Pendland S: Resveratrol inhibits growth of Helicobacter pylori in vitro. Amer J Gastroenterol 95:1849, 2000Google Scholar
  22. 22.
    Daroch F, Hoeneisen M, Gonzalez CL, Kawaguchi F, Salgado F, Solar H, Garcia A: In vitro antibacterial activity of Chilean red wines against Helicobacter pylori. Microbios 104: 79–85, 2001Google Scholar
  23. 23.
    Cover TL, Blaser MJ: Purification and characterization of the vacuolating toxin from Helicobacter pylori. J Biol Chem 267: 10570–10575, 1992PubMedGoogle Scholar
  24. 24.
    Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, Joshi SS, Pruess HG: Free radicals and grape seed proanthocyanidin extract: Importance in human health and disease prevention. Toxicology 148: 187–197, 2000Google Scholar
  25. 25.
    Chatterjee A, Yasmin T, Bagchi D, Stohs SJ: Inhibition of Helicobacter pylori in vitro by various berry extracts, with enhanced susceptibility to clarithromycin. Mol Cell Biochem 265: 19–26, 2004Google Scholar
  26. 26.
    Marshall BJ, Warren JR, Francis GJ, Langton SR, Goodwin CS, Blincow ED: Rapid urease test in the management of Campylobacter pyloridis-associated gastritis. Am J Gastroenterol 82: 200–210, 1987Google Scholar
  27. 27.
    Blaser MJ, Berg DE: Helicobacter pylori genetic diversity and risk of human disease. J Clin Invest 107: 767–773, 2001Google Scholar
  28. 28.
    Wilkinson M: Helicobacter pylori: An overview. Br J Biomed Sci 58: 59–60, 2001Google Scholar
  29. 29.
    Brown LM: Helicobacter pylori: Epidemiology and routes of transmission. Epidemiol Rev 22: 283–297, 2000Google Scholar
  30. 30.
    Leung WK, Graham DY: Clarithromycin for Helicobacter pylori infection. Expert Opin Pharmacother 1: 507–514, 2000Google Scholar
  31. 31.
    Aguilar GR, Ayala G, Fierros-Zarate G: Helicobacter pylori: Recent advances in the study of its pathogenicity and prevention. Salud Publica Mex 43: 237–247, 2001Google Scholar
  32. 32.
    Kozu T, Saito D: Helicobacter pylori and gastric disease. Rinsho Byori 49: 121–125, 2001Google Scholar
  33. 33.
    Wong WM, Gu Q, Wang WH, Fung FM, Berg DE, Lai KC, Xia HH, Hu WH, Chan CK, Chan AO, Yuen MF, Hui CK, Lam SK, Wong BC: Effects of primary metronidazole and clarithromycin resistance to Helicobacter pylori on omeprazole, metronidazole, and clarithromycin triple-therapy regimen in a region with high rates of metronidazole resistance. Clin Infect Dis 37: 882–889, 2003Google Scholar
  34. 34.
    Hao Q, Li Y, Zhang ZJ, Liu Y, Gao H: New mutation points in 23S rRNA gene associated with Helicobacter pylori resistance to clarithromycin in northeast China. World J Gastroenterol 10: 1075–1077, 2004Google Scholar
  35. 35.
    Elviss NC, Owen RJ, Xerry J, Walker AM, Davies K: Helicobacter pylori antibiotic resistance patterns and genotypes in adult dyspeptic patients from a regional population in North Wales. J Antimicrob Chemother 54: 435–440, 2004Google Scholar
  36. 36.
    Jung HK, Lee KE, Chu SH, Yi SY: Reactive oxygen species activity, mucosal lipoperoxidation and glutathione in Helicobacter pylori-infected gastric mucosa. J Gastroenterol Hepatol 16: 1336–1340, 2001Google Scholar
  37. 37.
    Ramarao N, Gray-Owen SD, Meyer TF: Helicobacter pylori induces but survives the extracellular release of oxygen radicals from professional phagocytes using its catalase activity. Mol Microbiol 38:103–113, 2000Google Scholar
  38. 38.
    Davies GR, Simmonds NJ, Stevens TR, Sheaff MT, Banatvala N, Laurenson IF, Blake DR, Rampton DS: Helicobacter pylori stimulates antral mucosal reactive oxygen metabolite production in vivo. Gut 35: 179–185, 1994Google Scholar
  39. 39.
    Salim AS: The relationship between Helicobacter pylori and oxygen-derived free radicals in the mechanism of duodenal ulceration. Intern Med 32: 359–364, 1993Google Scholar
  40. 40.
    Yang J, Kato K, Noguchi K, Dairaku N, Koike T, Iijima K, Imatani A, Sekine H, Ohara S, Sasano H, Shimosegawa T: Tochu (Eucommia ulmoides) leaf extract prevents ammonia and vitamin C deficiency induced gastric mucosal injury. Life Sci 73: 3245–3256, 2003Google Scholar
  41. 41.
    Haruma K, Ito M: Clinical significance of mucosal-protective agents: Acid, inflammation, carcinogenesis and rebamipide. Aliment Pharmacol Ther 18: 153–159, 2003Google Scholar
  42. 42.
    Yanagawa Y, Yamamoto Y, Hara Y, Shimamura T: A combination effect of epigallocatechin gallate, a major component of green tea catechins, with antibiotics on Helicobacter pylori growth in vitro. Curr Microbiol 47: 244–249, 2003Google Scholar
  43. 43.
    Berspalov VG, Shcherbakov AM, Kalinovskii VP, Novik VI, Chepik OF, Aleksandrov VA, Sobenin IA, Orekhov AN: Study of the antioxidant drug “Karinat” in patients with chronic atrophic gastritis. Vopr Onkol 50: 81–85, 2004Google Scholar
  44. 44.
    Wong JE, Ito Y, Correa P, Peeters KC, van de Velde CJ, Sasako M, Macdonald J: Therapeutic strategies in gastric cancer. J Clin Oncol 21(23 Suppl): 267S–269S, 2003Google Scholar
  45. 45.
    Correa P, Piazuelo MB, Camargo MC: The future of gastric cancer prevention. Gastric Cancer 7: 9–16, 2004Google Scholar
  46. 46.
    Mahady GB, Pendland SL, Chadwick LR: Resveratrol and red wine extracts inhibit the growth of cagA+ strains of Helicobacter pylori in vitro. Am J Gastronterol 98: 1440–1441, 2003Google Scholar
  47. 47.
    Sato M, Maulik G, Bagchi D, Das DK: Myocardial protection by Protykin®, a novel extract of trans-resveratrol and emodin. Free Radic Res 32: 135–144, 2000Google Scholar
  48. 48.
    Zhang H, Wakisaka N, Maeda O, Yamamoto T: Vitamin C inhibits the growth of a bacterial risk factor for gastric carcinoma: Helicobacter pylori. Cancer 80: 1897–1903, 1997Google Scholar
  49. 49.
    Wang X, Willen R, Wadström T: Astaxanthin-rich algal meal and vitamin C inhibit Helicobacter pylori infection in BALB/cA mice. Antimicrob Agents Chemother 44: 2452–2457, 2000CrossRefGoogle Scholar
  50. 50.
    Sobala GM, Schorah CJ, Shires S, Lynch DA, Gallacher B, Dixon MF, Axon AT: Effect of eradication of Helicobacter pylori on gastric juice ascorbic acid concentrations. Gut 34: 1038–1041, 1993Google Scholar
  51. 51.
    Banerjee S, Hawksby C, Miller S, Dahill S, Beattie AD, McColl KE: Effect of Helicobacter pylori and its eradication on gastric juice ascorbic acid. Gut 35: 317–322, 1994Google Scholar
  52. 52.
    Drake IM, Davies MJ, Mapstone NP, Dixon MF, Schorah CJ, White KL, Chalmers DM, Axon AT: Ascorbic acid may protect against human gastric cancer by scavenging mucosal oxygen radicals. Carcinogenesis 17: 559–562, 1996Google Scholar
  53. 53.
    Everett SM, Drake IM, White KL, Mapstone NP, Chalmers DM, Schorah CJ, Axon AT: Antioxidant vitamin supplements do not reduce reactive oxygen species activity in Helicobacter pylori gastritis in the short term. Br J Nutr 87: 3–11, 2002Google Scholar
  54. 54.
    Chuang CH, Sheu BS, Huang AH, Yang HB, Wu JJ: Vitamin C and E supplements to lansoprazole-amoxicillin-metronidazole triple therapy may reduce the eradication rate of metronidazole-susceptible Helicobacter pylori infection. Helicobacter 7: 310–316, 2002Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Archana Chatterjee
    • 1
    • 2
    • 3
  • Debasis Bagchi
    • 2
  • Taharat Yasmin
    • 2
  • Sidney J. Stohs
    • 2
  1. 1.Department of Pediatrics, Division of Pediatric Infectious DiseasesCreighton University Medical CenterOmahaUSA
  2. 2.Department of Pharmacy SciencesCreighton University Medical CenterOmahaUSA
  3. 3.Department of PediatricsCreighton University Health Science CenterOmahaUSA

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