Polysorbate 80 and Helicobacter pylori: a microbiological and ultrastructural study

BMC Microbiology

Elena Moretti⁴  Adriano Spreafico²  Giulia Collodel⁴  Annalisa Santucci³  Giovanni Cavallo⁵  Roberto Marcolongo⁶  Natale Figura¹ 

[1] Department of Internal Medicine, University of Siena, Siena, Italy;Department of Clinical Medicine and Immunological Sciences, University of Siena, Siena, Italy;Biotechnology Department, University of Siena, Siena, Italy;Department of Biomedical Sciences, Applied Biology Section, University of Siena, Siena, Italy;Qi s.r.l, Pomezia, Rome, Italy;Centre for Biochemical and Clinical Study of Rheumatic Diseases, Siena, Italy

关键词: Transmission electron microscopy;    Polysorbate 80;    Chemoresistances;    Antibiotics;    Bacteria;   

Others  :  1221746
DOI  :  10.1186/1471-2180-12-217

 received in 2012-05-11, accepted in 2012-09-14,  发布年份 2012

【 摘 要 】

Background

The frequent occurrence of chemoresistant strains reduces the chances of eradication of H. pylori infection and prompted the investigation of non-antibiotic substances active against this organism. Some surfactants enhance the effectiveness of antibiotics for their permeabilizing properties towards bacteria. We examined the antimicrobial activity to H. pylori of the surfactant polysorbate 80, used alone and in association with amoxicillin, clarithromycin, metronidazole, levofloxacin and tetracycline. We also aimed to study the ultrastructural alterations caused upon H. pylori by polysorbate 80, alone and in combination with antibiotics. Twenty-two H. pylori strains were tested using the broth dilution method. After incubation, broth from each dilution was subcultured onto agar enriched with foetal bovine serum to determine the minimum bactericidal concentration (MBC). Synergistic effect of polysorbate 80 with antibiotics was investigated by the broth dilution and disc diffusion techniques. Ultrastructural alterations of organisms treated with polysorbate 80, alone and in association with antibiotics were analyzed by transmission electron microscopy.

Results

MBCs of polysorbate 80 ranged from 2.6 (1.1) μg/ml to 32 (0) μg/ml. Polysorbate 80 exerted a synergistic effect when associated with metronidazole and clarithromycin: polysorbate 80 and metronidazole MBCs decreased by ≥ 4 fold; clarithromycin MBCs for two resistant strains decreased by 20 and 1000 times. The principal alteration caused by polysorbate 80 consisted in the detachment of the outer membrane of bacteria.

Conclusions

The bactericidal activity of polysorbate 80 and the synergistic effect of the association with metronidazole and clarithromycin could be useful in the treatment of H. pylori infection.

【 授权许可】

   
2012 Figura et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Telford JL, Covacci A, Rappuoli R, Ghiara P: Immunobiology of Helicobacter pylori infection. Curr Opin Immunol 1997, 9:498-503.
• [2]Walker MM, Crabtree JE: Helicobacter pylori infection and the pathogenesis of duodenal ulceration. Ann N Y Acad Sci 1998, 859:96-111.
• [3]Mégraud F: Basis for the management of drug-resistant Helicobacter pylori infection. Drugs 2004, 64:1893-1904.
• [4]Kusters JG, van Vliet AH, Kuipers EJ: Pathogenesis of Helicobacter pylori infection. Clin Microbiol Rev 2006, 19:449-490.
• [5]O’Connor A, Gisbert JP, McNamara D, O'Morain C: Treatment of Helicobacter pylori infection 2010. Helicobacter 2010, 15:46-52.
• [6]Gatta L, Vakil N, Leandro G, Di Mario F, Vaira D: Sequential therapy or triple therapy for Helicobacter pylori infection: systematic review and meta-analysis of randomized controlled trials in adults and children. Am J Gastroenterol 2009, 104:3069-3079.
• [7]Malfertheiner P, Bazzoli F, Delchier JC, Celiñski K, Giguère M, Rivière M, Mégraud F, Pylera Study Group: Helicobacter pylori eradication with a capsule containing bismuth subcitrate potassium, metronidazole, and tetracycline given with omeprazole versus clarithromycin-based triple therapy: a randomised, open-label, non-inferiority, phase 3 trial. Lancet 2011, 377:905-913.
• [8]Parente F, Cucino C, Bianchi PG: Treatment options for patients with Helicobacter pylori infection resistant to one or more eradication attempts. Dig Liver Dis 2003, 35:523-528.
• [9]De Francesco V, Giorgio F, Hassan C, Manes G, Vannella L, Panella C, Ierardi E, Zullo A: Worldwide H. pylori antibiotic resistance: a systematic review. J Gastrointestin Liver Dis 2010, 19:409-414.
• [10]Selgrad M, Malfertheiner P: Treatment of Helicobacter pylori. Curr Opin Gastroenterol 2011, 27:565-570.
• [11]Fischbach L, Evans EL: Meta-analysis: the effect of antibiotic resistance status on the efficacy of triple and quadruple first-line therapies for Helicobacter pylori. Aliment Pharmacol Ther 2007, 26:343-357.
• [12]Laine L, Fennerty MB, Osato M, Sugg J, Suchower L, Probst P, Levine JG: Esomeprazole-based Helicobacter pylori eradication therapy and the effect of antibiotic resistance: results of three US multicenter, double-blind trials. Am J Gastroenterol 2000, 95:3393-3398.
• [13]Treiber G, Malfertheiner P, Klotz U: Treatment and dosing of Helicobacter pylori infection: when pharmacology meets clinic. Expert Opin Pharmacother 2007, 8:329-350.
• [14]Vítor JMB, Vale FF: Alternative therapies for Helicobacter pylori: probiotics and phytomedicine. FEMS Immunol Med Microbiol 2011, 63:153-164.
• [15]Zhang H, Shen Y, Weng P, Zhao G, Feng F, Zheng X: Antimicrobial activity of a food-grade fully dilutable microemulsion against Escherichia coli and Staphylococcus aureus. Int J Food Microbiol 2009, 135:211-215.
• [16]Stoops JK, Arora R, Armitage L, Song L, Blackburn MR, Krueger GR, Risin SA: Certain surfactants show promise in the therapy of pulmonary tuberculosis. In Vivo 2010, 24:687-694.
• [17]Huesca M, Gold B, Sherman P, Lewin P, Lingwood C: Therapeutics used to alleviate peptic ulcers inhibit H. pylori receptor binding in vitro. Zentralbl Bakteriol 1993, 280:244-252.
• [18]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 2004, 10:1088-1094.
• [19]Björkholm B, Sjolund M, Falk PG, Berg OG, Engstrand L, Andersson DI: Mutation frequency and biological cost of antibiotic resistance in Helicobacter pylori. Proc Natl Acad Sci USA 2001, 98:14607-14612.
• [20]Dorer MS, Fero J, Salama NR: DNA damage triggers genetic exchange in Helicobacter pylori. PLoS Pathog 2010, 6:e1001026.
• [21]Fischer W, Windhager L, Rohrer S, Karnholz A, Hoffmann R, Zimmer R, Haas R: Strain-specific genes of Helicobacter pylori: genome evolution driven by a novel type IV secretion system and genomic island transfer. Nucleic Acids Res 2010, 38:6089-6101.
• [22]Ndip RN, Malange Tarkang AE, Mbullah SM, Luma HN, Malongue A, Ndip LM, Nyongbela K, Wirmum C, Efange SM: In vitro anti-Helicobacter pylori activity of extracts of selected medicinal plants from North West Cameroon. J Ethnopharmacol 2007, 114:452-457.
• [23]Williams J, Odum J, Lewis RW, Brady AM: The oral administration of polysorbate 80 to the immature female rat does not increase uterine weight. Toxicol Lett 1997, 91:19-24.
• [24]Ema M, Hara H, Matsumoto M, Hirata-Koizumi M, Hirose A, Kamata E: Evaluation of developmental neurotoxicity of polysorbate 80 in rats. Reprod Toxicol 2008, 25:89-99.
• [25]Parker H, Keenan JI: Composition and function of Helicobacter pylori outer membrane vesicles. Microbes Infect 2012, 14:9-16.
• [26]Armstrong JA, Wee SH, Goodwin CS, Wilson DH: Response of Campylobacter pyloridis to antibiotics, bismuth and an acid-reducing agent in vitro–an ultrastructural study. J Med Microbiol 1987, 24:343-350.
• [27]Chey WD, Wong BC: American College of Gastroenterology guideline on the management of Helicobacter pylori infection. Am J Gastroenterol 2007, 102:1808-1825.
• [28]Marais A, Bilardi C, Cantet F, Mendz GL, Mégraud F: Characterization of the genes rdxA and frxA involved in metronidazole resistance in Helicobacter pylori. Res Microbiol 2003, 154:137-144.
• [29]Tsugawa H, Suzuki H, Muraoka H, Ikeda F, Hirata K, Matsuzaki J, Saito Y, Hibi T: Enhanced bacterial efflux system is the first step to the development of metronidazole resistance in Helicobacter pylori. Biochem Biophys Res Commun 2011, 404:656-660.
• [30]van Amsterdam K, Bart A, van der Ende A: A Helicobacter pylori TolC efflux pump confers resistance to metronidazole. Antimicrob Agents Chemother 2005, 49:1477-1482.
• [31]Liu ZQ, Zheng PY, Yang PC: Efflux pump gene hefA of Helicobacter pylori plays an important role in multidrug resistance. World J Gastroenterol 2008, 14:5217-5222.
• [32]Paulsen IT, Chen J, Nelson KE, Saier MH Jr: Comparative genomics of microbial drug efflux systems. J Mol Microbiol Biotechnol 2001, 3:145-150.
• [33]Johnson JM, Church GM: Alignment and structure prediction of divergent protein families: periplasmic and outer membrane proteins of bacterial efflux pumps. J Mol Biol 1999, 287:695-715.
• [34]Delcour AH: Outer membrane permeability and antibiotic resistance. Biochim Biophys Acta 2009, 1794:808-816.
• [35]Vaara M: Agents that increase the permeability of the outer membrane. Microbiol Rev 1992, 56:395-411.
• [36]Savage PB: Multidrug-resistant bacteria: overcoming antibiotic permeability barriers of gram-negative bacteria. Ann Med 2001, 33:167-171.
• [37]Mahachai V, Sirimontaporn N, Tumwasorn S, Thong-Ngam D, Vilaichone RK: Sequential therapy in clarithromycin-sensitive and –resistant Helicobacter pylori based on polymerase chain reaction molecular test. J Gastroenterol Hepatol 2011, 26:825-828.
• [38]Bina JE, Alm RA, Uria-Nickelsen M, Thomas SR, Trust TJ, Hancock RE: Helicobacter pylori uptake and efflux: basis for intrinsic susceptibility to antibiotics in vitro. Antimicrob Agents Chemother 2000, 44:248-254.
• [39]Nikaido H: Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 2003, 67:593-656.