Skip to main content
SpringerLink
Log in

Lactobacillus fermentum, a probiotic capable to release glutathione, prevents colonic inflammation in the TNBS model of rat colitis

  • Original Article
  • Published:
International Journal of Colorectal Disease Aims and scope Submit manuscript

Abstract

Background and aims

Inflammatory bowel disease is associated with intestinal oxidative stress. In the present study we test the preventative effect of Lactobacillus fermentum, a probiotic that produces per se glutathione, in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis.

Methods

Colitis was induced in rats by intracolonic administration of 10 mg of TNBS dissolved in 0.25 ml of 50% ethanol. L. fermentum was administered orally (5×108 CFU suspended in 0.5 ml of skim milk) to a group of rats for 3 weeks, starting 2 weeks before colitis induction. Colonic damage was evaluated both histologically and biochemically, and the colonic luminal contents were used for bacterial studies as well as for short chain fatty acid (SCFA) production.

Results

L. fermentum treatment resulted in an amelioration of the inflammatory response in colitic rats as evidenced histologically and by a significant reduction of colonic MPO activity (P<0.05). The probiotic partially counteracted the colonic glutathione depletion induced by the inflammatory process. In addition, probiotic-treated colitic rats showed significant lower colonic tumour necrosis factor (TNF)α levels (P<0.01) and inducible nitric oxide synthase (iNOS) expression when compared to non-treated rats. Finally, the probiotic induced growth of Lactobacilli species and production of SCFA in colonic contents in comparison with control colitic rats.

Conclusion

Administration of the probiotic L. fermentum facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with increased levels of glutathione as well as with amelioration of the production of some of the mediators involved in the inflammatory response of the intestine, such as TNFα and NO.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Fiocchi C (1998) Inflammatory bowel disease: aetiology and pathogenesis. Gastroenterology 115:182–205

    Article  PubMed  CAS  Google Scholar 

  2. Shanahan F (2000) Probiotics and inflammatory bowel disease: is there a scientific rationale? Inflamm Bowel Dis 6:107–115

    Article  PubMed  CAS  Google Scholar 

  3. Chung PY, Peppercorn MA (1999) Antibiotics in inflammatory bowel disease. Drugs Today 35:89–103

    CAS  PubMed  Google Scholar 

  4. Taurog JD, Richardson JA, Croft JT, Simmons WA, Zhou M, Fernandez-Sueiro JL, Balish E, Hammer RE (1994) The germ-free state prevents development of gut and joint inflammatory disease in HLA-B27 transgenic rats. J Exp Med 180:2359–2364

    Article  PubMed  CAS  Google Scholar 

  5. Venturi A, Gionchetti P, Rizzello F, Johansson R, Zucconi E, Brigidi P, Matteuzzi D, Campieri M (1999) Impact on the composition of the faecal flora by a new probiotic preparation: preliminary data on maintenance treatment of patients with ulcerative colitis. Aliment Pharmacol Ther 13:1103–1108

    Article  PubMed  CAS  Google Scholar 

  6. Rembacken BJ, Snelling AM, Hawkey PM, Chalmers DM, Axon AT (1999) Non-pathogenic Escherichia coli versus mesalazine for the treatment of ulcerative colitis: a randomised trial. Lancet 354:635–639

    Article  PubMed  CAS  Google Scholar 

  7. Gionchetti P, Rizzello F, Helwig U, Venturi A, Lammers KM, Brigidi P, Vitali B, Poggioli G, Miglioli M, Campieri M (2003) Prophylaxis of pouchitis onset with probiotic therapy: a double-blind, placebo-controlled trial. Gastroenterology 124:1202–1209

    Article  PubMed  Google Scholar 

  8. Gionchetti P, Rizzello F, Venturi A, Brigidi P, Matteuzzi D, Bazzocchi G, Poggioli G, Miglioli M, Campieri M (2000) Oral bacteriotherapy as maintenance treatment in patients with chronic pouchitis: a double-blind, placebo-controlled trial. Gastroenterology 119:305–309

    Article  PubMed  CAS  Google Scholar 

  9. Tannock GW, Munro K, Harmsen HJ, Welling GW, Smart J, Gopal PK (2000) Analysis of the fecal microflora of human subjects consuming a probiotic product containing Lactobacillus rhamnosus DR20. Appl Environ Microbiol 66:2578–2588

    Article  PubMed  CAS  Google Scholar 

  10. Ulisse S, Gionchetti P, D’Alo S, Russo FP, Pesce I, Ricci G, Rizzello F, Helwig U et al (2001) Expression of cytokines, inducible nitric oxide synthase, and matrix metalloproteinases in pouchitis: effects of probiotic treatment. Am J Gastroenterol 96:2691–2699

    Article  PubMed  CAS  Google Scholar 

  11. Schultz M, Linde HJ, Lehn N, Zimmermann K, Grossmann J, Falk W, Scholmerich J (2003) Immunomodulatory consequences of oral administration of Lactobacillus rhamnosus strain GG in healthy volunteers. J Dairy Res 70:165–173

    Article  PubMed  CAS  Google Scholar 

  12. Madsen K, Cornish A, Soper P, McKaigney C, Jijon H, Yachimec C, Doyle J, Jewell L, De Simone C (2001) Probiotic bacteria enhance murine and human intestinal epithelial barrier function. Gastroenterology 121:580–591

    Article  PubMed  CAS  Google Scholar 

  13. Otte JM, Podolsky DK (2004) Functional modulation of enterocytes by gram-positive and gram-negative microorganisms. Am J Physiol 286:G613–G626

    CAS  Google Scholar 

  14. Steidler L, Hans W, Schotte L, Neirynck S, Obermeier F, Falk W, Fiers W, Remaut E (2000) Treatment of murine colitis by Lactococcus lactis secreting interleukin-10. Science 289:1352–1355

    Article  PubMed  CAS  Google Scholar 

  15. Vandenbroucke K, Hans W, Van Huysse J, Neirynck S, Demetter P, Remaut E, Rottiers P, Steidler L (2004) Active delivery of trefoil factors by genetically modified Lactococcus lactis prevents and heals acute colitis in mice. Gastroenterology 127:502–513

    Article  PubMed  CAS  Google Scholar 

  16. Lamine F, Fioramonti J, Bueno L, Nepveu F, Cauquil E, Lobysheva I, Eutamene H, Theodorou V (2004) Nitric oxide released by Lactobacillus farciminis improves TNBS-induced colitis in rats. Scand J Gastroenterol 39:37–45

    Article  PubMed  CAS  Google Scholar 

  17. Pavlick KP, Laroux FS, Fuseler J, Wolf RE, Gray L, Hoffman J, Grisham MB (2002) Role of reactive metabolites of oxygen and nitrogen in inflammatory bowel disease. Free Radic Biol Med 33:311–322

    Article  PubMed  CAS  Google Scholar 

  18. Buffinton GD, Doe WF (1995) Depleted mucosal antioxidant defences in inflammatory bowel disease. Free Radic Biol Med 19:911–918

    Article  PubMed  CAS  Google Scholar 

  19. Galvez J, Coelho G, Crespo ME, Cruz T, Rodriguez-Cabezas ME, Concha A, Gonzalez M, Zarzuelo A (2001) Intestinal anti-inflammatory activity of morin on chronic experimental colitis in the rat. Aliment Pharmacol Ther 15:2027–2039

    Article  PubMed  CAS  Google Scholar 

  20. Camuesco D, Comalada M, Rodriguez-Cabezas ME, Nieto A, Lorente MD, Concha A, Zarzuelo A, Galvez J (2004) The intestinal anti-inflammatory effect of quercitrin is associated with an inhibition in iNOS expression. Br J Pharmacol, DOI 10.1038/sj.bjp.0705941

  21. Gonzalez R, Sanchez de Medina F, Galvez J, Rodriguez-Cabezas ME, Duarte J, Zarzuelo A (2001) Dietary vitamin E supplementation protects the rat large intestine from experimental inflammation. Int J Vitam Nutr Res 71:243–250

    Article  PubMed  CAS  Google Scholar 

  22. Travis SPL, Jewel DP (1994) Salicylates for ulcerative colitis—their mode of action. Pharmacol Ther 63:135–161

    Article  PubMed  CAS  Google Scholar 

  23. Rachmilewitz D, Karmeli F, Okon E (1995) Sulfhydryl blocker-induced rat colonic inflammation is ameliorated by inhibition of nitric oxide synthase. Gastroenterology 109:98–106

    Article  PubMed  CAS  Google Scholar 

  24. Loguercio C, D’Argenio G, Delle Cave M, Cosenza V, Della Valle N, Mazzacca G, Del Vecchio Blanco C (2003) Glutathione supplementation improves oxidative damage in experimental colitis. Dig Liver Dis 35:635–641

    Article  PubMed  CAS  Google Scholar 

  25. Ardite E, Sans M, Panes J, Romero FJ, Pique JM, Fernandez-Checa JC (2000) Replenishment of glutathione levels improves mucosal function in experimental colitis. Lab Invest 80:735–744

    PubMed  CAS  Google Scholar 

  26. Jurjus AR, Khoury NN, Reimund JM (2004) Animal models of inflammatory bowel disease. J Pharmacol Toxicol Methods 50:81–92

    Article  PubMed  CAS  Google Scholar 

  27. Martin R, Langa S, Reviriego C, Jiminez E, Marin ML, Xaus J, Fernandez L, Rodriguez JM (2003) Human milk is a source of lactic acid bacteria for the infant gut. J Pediatr 143:754–758

    Article  PubMed  CAS  Google Scholar 

  28. Morris GP, Beck PL, Herridge W, Depew W, Szewczuk MR, Wallace JL (1989) Hapten induced model of chronic inflammation and ulceration in rat colon. Gastroenterology 96:795–803

    PubMed  CAS  Google Scholar 

  29. Bell CJ, Gall DG, Wallace JL (1995) Disruption of colonic electrolyte transport in experimental colitis. Am J Physiol 268:G622–G630

    PubMed  CAS  Google Scholar 

  30. Camuesco D, Peran L, Comalada M, Nieto A, Di Stasi LC, Rodriguez-Cabezas ME, Concha A, Zarzuelo A, Galvez J (2005) Preventative effects of lactulose in the trinitrobenzenesulphonic acid model of rat colitis. Inflamm Bowel Dis 11:265–271

    Article  PubMed  Google Scholar 

  31. Krawisz JE, Sharon P, Stenson WF (1984) Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology 87:1344–1350

    PubMed  CAS  Google Scholar 

  32. Anderson ME (1985) Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol 113:548–555

    Article  PubMed  CAS  Google Scholar 

  33. Newton GL, Arnold K, Price MS, Sherrill C, Delcardayre SB, Aharanowitz Y, Cohen G, Davies J et al (1996) Distribution of thiols in microorganisms: mycothiol is a mayor thiol in most Actinomycetes. J Bacteriol 178:1990–1995

    PubMed  CAS  Google Scholar 

  34. Fahey RC, Brown WC, Adams WB, Worsham MB (1978) Occurrence of glutathione in bacteria. J Bacteriol 133:1126–1129

    PubMed  CAS  Google Scholar 

  35. Galvez J, Garrido M, Rodriguez-Cabezas ME, Ramis I, Sanchez de Medina F, Merlos M, Zarzuelo A (2003) The intestinal anti-inflammatory activity of UR-12746S on reactivated experimental colitis is mediated through downregulation of cytokine production. Inflamm Bowel Dis 9:363–371

    Article  PubMed  Google Scholar 

  36. Rodriguez-Cabezas ME, Galvez J, Lorente MD, Concha A, Camuesco D, Azzouz S, Osuna A, Redondo L, Zarzuelo A (2002) Dietary fiber down-regulates colonic tumor necrosis factor alpha and nitric oxide production in trinitrobenzenesulfonic acid-induced colitic rats. J Nutr 132:3263–3271

    PubMed  CAS  Google Scholar 

  37. Miralles-Barrachina O, Savoye G, Belmonte-Zalar L, Hochain P, Ducrotte P, Hecketsweiler B, Lerebours E, Dechelotte P (1999) Low levels of glutathione in endoscopic biopsies of patients with Crohn’s colitis: the role of malnutrition. Clin Nutr 18:313–317

    Article  PubMed  CAS  Google Scholar 

  38. Schultz M, Sartor RB (2000) Probiotics and inflammatory bowel diseases. Am J Gastroenterol 95:S19–S21

    Article  Google Scholar 

  39. Veljaca M, Lesch CA, Pllana R, Sanchez B, Chan K, Guglietta A (1995) BPC-15 reduces trinitrobenzene sulfonic-induced colonic damage in rats. J Pharmacol Exp Ther 272:417–422

    PubMed  CAS  Google Scholar 

  40. Ajuebor MN, Zagorski J, Kunkel SL, Strieter RM, Hogaboam CM (2004) Contrasting roles for CXCR2 during experimental colitis. Exp Mol Pathol 76:1–8

    Article  PubMed  CAS  Google Scholar 

  41. Grisham MB, Olkmer C, Tso P, Yamada T (1991) Metabolism of trinitrobenzene sulfonic acid by rat colon produces reactive oxygen species. Gastroenterology 101:540–547

    PubMed  CAS  Google Scholar 

  42. Wu G, Fang YZ, Yang S, Lupton JR, Turner ND (2004) Glutathione metabolism and its implications for health. J Nutr 134:489–492

    PubMed  CAS  Google Scholar 

  43. Guo X, Wang WP, Ko JK, Cho CH (1999) Involvement of neutrophils and free radicals in the potentiating effects of passive cigarette smoking on inflammatory bowel disease in rats. Gastroenterology 117:884–892

    Article  PubMed  CAS  Google Scholar 

  44. Rachmilewitz D, Karmeli F, Okon E, Bursztyn M (1995) Experimental colitis is ameliorated by inhibition of nitric oxide synthase activity. Gut 37:247–255

    PubMed  CAS  Google Scholar 

  45. Hogaboam CM, Jacobson K, Collins SM Blennerhassett MG (1995) The selective beneficial effects of nitric oxide inhibition in experimental colitis. Am J Physiol 268:G673–G684

    PubMed  CAS  Google Scholar 

  46. Kimura H, Miura S, Shigematsu T, Ohkubo N, Tsuzuki Y, Kurose I, Higuchi H, Akiba Y et al (1997) Increased nitric oxide production and inducible nitric oxide synthase activity in colonic mucosa of patients with active ulcerative colitis and Crohn’s disease. Dig Dis Sci 42:1047–1054

    Article  PubMed  CAS  Google Scholar 

  47. Ikeda I, Kasajima T, Ishiyama S, Shimojo T, Takeo Y, Nishikawa T, Kameoka S, Hiroe M, Mitsunaga A (1997) Distribution of inducible nitric oxide synthase in ulcerative colitis. Am J Gastroenterol 92:1339–1341

    PubMed  CAS  Google Scholar 

  48. McCafferty DM, Miampamba M, Sihota E, Sharkey KA, Kubes P (1999) Role of inducible nitric oxide synthase in trinitrobenzene sulphonic acid induced colitis in mice. Gut 45:864–873

    Article  PubMed  CAS  Google Scholar 

  49. Van Deventer SJH (1997) Tumor necrosis factor and Crohn’s disease. Gut 40:443–448

    PubMed  Google Scholar 

  50. Rutgeerts P, Van Assche G, Vermeire S (2004) Optimizing anti-TNF treatment in inflammatory bowel disease. Gastroenterology 126:1593–1610

    Article  PubMed  CAS  Google Scholar 

  51. Borruel N, Carol M, Casellas F, Antolin M, de Lara F, Espin E, Naval J, Guarner F, Malagelada JR (2002) Increased mucosal tumour necrosis factor alpha production in Crohn’s disease can be downregulated ex vivo by probiotic bacteria. Gut 51:659–664

    Article  PubMed  CAS  Google Scholar 

  52. Schottelius AJ, Baldwin AS Jr (1999) A role for transcription factor NF-kappa B in intestinal inflammation. Int J Colorectal Dis 14:18–28

    Article  PubMed  CAS  Google Scholar 

  53. Segain JP, Raingeard de la Bletiere D, Bourreille A, Leray V, Gervois N, Rosales C, Ferrier L, Bonnet C et al (2000) Butyrate inhibits inflammatory responses through NFκB inhibition: implications for Crohn’s disease. Gut 47:397–403

    Article  PubMed  CAS  Google Scholar 

  54. Inan MS, Rasoulpour RJ, Yin L, Hubbard AK, Rosenberg DW, Giardina C (2000) The luminal short-chain fatty acid butyrate modulates NF-κB activity in a human colonic epithelial cell line. Gastroenterology 118:724–734

    Article  PubMed  CAS  Google Scholar 

  55. Rogler G, Brand K, Vogl D, Page S, Hofmeister R, Andus T, Knuechel R, Baeuerle PA et al (1998) Nuclear factor kappaB is activated in macrophages and epithelial cells of inflamed intestinal mucosa. Gastroenterology 115:357–369

    Article  PubMed  CAS  Google Scholar 

  56. Garcia-Lafuente A, Antolin M, Guarner F, Crespo E, Salas A, Forcada P, Laguarda M, Gavalda J et al (1997) Incrimination of anaerobic bacteria in the induction of experimental colitis. Am J Physiol 272:G10–G15

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the Spanish Ministry of Science and Technology (SAF2002-02592) and by Instituto de Salud ‘Carlos III’ (PI021732), with funds from the European Union, and by Junta de Andalucia (CTS 164). Mònica Comalada is a recipient of Juan de la Cierva Program from Spanish Ministry of Science and Technology. Laura Perán is a recipient from Puleva Foundation (Spain).

Author information

Authors and Affiliations

  1. Department of Pharmacology, School of Pharmacy, University of Granada, Campus Universitario La Cartuja s/n, 18071, Granada, Spain

    Laura Peran, Desiree Camuesco, Monica Comalada, Antonio Zarzuelo & Julio Galvez

  2. Health and Progress Foundation, Granada, Spain

    Ana Nieto

  3. Department of Pathology, Hospital Universitario Virgen de las Nieves, Granada, Spain

    Angel Concha

  4. Department of Immunology and Animal Sciences, Puleva Biotech, S.A., Granada, Spain

    José Luis Adrio, Mónica Olivares & Jordi Xaus

Authors
  1. Laura Peran
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Desiree Camuesco
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Monica Comalada
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana Nieto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Angel Concha
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. José Luis Adrio
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mónica Olivares
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jordi Xaus
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Antonio Zarzuelo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Julio Galvez
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Julio Galvez.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Peran, L., Camuesco, D., Comalada, M. et al. Lactobacillus fermentum, a probiotic capable to release glutathione, prevents colonic inflammation in the TNBS model of rat colitis. Int J Colorectal Dis 21, 737–746 (2006). https://doi.org/10.1007/s00384-005-0773-y

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00384-005-0773-y

Keywords

Search

Navigation