Molecular and Cellular Biochemistry

, Volume 362, Issue 1–2, pp 43–53 | Cite as

Antinociceptive effect of VSL#3 on visceral hypersensitivity in a rat model of irritable bowel syndrome: a possible action through nitric oxide pathway and enhance barrier function

  • Cong Dai
  • Stefano Guandalini
  • De-Hui Zhao
  • Min Jiang


Irritable bowel syndrome (IBS) is a functional bowel disorder characterized by visceral hypersensitivity and altered bowel function. There are increasing evidences suggested that VSL#3 probiotics therapy has been recognized as an effective method to relieve IBS-induced symptoms. The aim of this study was to examine the effects of VSL#3 probiotics on visceral hypersensitivity (VH), nitric oxide (NO), fecal character, colonic epithelium permeability, and tight junction protein expression. IBS model was induced by intracolonic instillation of 4% acetic acid and restraint stress in rats. After subsidence of inflammation on the seventh experimental day, the rats were subjected to rectal distension, and then the abdominal withdrawal reflex and the number of fecal output were measured, respectively. Also, colonic permeability to Evans blue was measured in vivo, and tight junction protein expression was studied by immunohistochemistry and immunoblotting method. Rats had been pretreated with VSL#3 or aminoguanidine (NOS inhibitor) or VSL#3+ aminoguanidine before measurements. The rats at placebo group showed hypersensitive response to rectal distension (P < 0.05) and defecated more stools than control rats (P < 0.05), whereas VSL#3 treatment significantly attenuated VH and effectively reduced defecation. Aminoguanidine reduced the protective effects of VSL#3 on VH. A pronounced increase in epithelial permeability and decreased expression of tight junction proteins (occludin, ZO-1) in placebo group were prevented by VSL#3, but not aminoguanidine. VSL#3 treatment reduce the hypersensitivity, defecation, colonic permeability and increase the expression of tight junction proteins (occludin, ZO-1). As the part of this effect was lowered by NOS inhibitor, NO might play a role in the protective effect of VSL#3 to some extent.


Irritable bowel syndrome Probiotic VSL#3 Intestinal barrier Tight junction Visceral hypersensitivity Nitric oxide 


  1. 1.
    Quigley EM, Flourie B (2007) Probiotics and irritable bowel syndrome: a rationale for their use and an assessment of the evidence to date. Neurogastroenterol Motil 19(3):166–172. doi: 10.1111/j.1365-2982.2006.00879.x PubMedCrossRefGoogle Scholar
  2. 2.
    McCarthy J, O’Mahony L, O’Callaghan L, Sheil B, Vaughan EE, Fitzsimons N, Fitzgibbon J, O’Sullivan GC, Kiely B, Collins JK, Shanahan F (2003) Double blind, placebo controlled trial of two probiotic strains in interleukin 10 knockout mice and mechanistic link with cytokine balance. Gut 52(7):975–980PubMedCrossRefGoogle Scholar
  3. 3.
    O’Mahony L, McCarthy J, Kelly P, Hurley G, Luo F, Chen K, O’Sullivan GC, Kiely B, Collins JK, Shanahan F, Quigley EM (2005) Lactobacillus and bifidobacterium in irritable bowel syndrome: symptom responses and relationship to cytokine profiles. Gastroenterology 128(3):541–551PubMedCrossRefGoogle Scholar
  4. 4.
    Verdu EF, Bercik P, Verma-Gandhu M, Huang XX, Blennerhassett P, Jackson W, Mao Y, Wang L, Rochat F, Collins SM (2006) Specific probiotic therapy attenuates antibiotic induced visceral hypersensitivity in mice. Gut 55(2):182–190. doi: 10.1136/gut.2005.066100 PubMedCrossRefGoogle Scholar
  5. 5.
    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(9179):635–639PubMedCrossRefGoogle Scholar
  6. 6.
    Gionchetti P, Rizzello F, Venturi A, Campieri M (2000) Probiotics in infective diarrhoea and inflammatory bowel diseases. J Gastroenterol Hepatol 15(5):489–493PubMedCrossRefGoogle Scholar
  7. 7.
    Sullivan A, Nord CE (2005) Probiotics and gastrointestinal diseases. J Intern Med 257(1):78–92. doi: 10.1111/j.1365-2796.2004.01410.x PubMedCrossRefGoogle Scholar
  8. 8.
    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(1):37–45PubMedCrossRefGoogle Scholar
  9. 9.
    Isolauri E, Majamaa H, Arvola T, Rantala I, Virtanen E, Arvilommi H (1993) Lactobacillus casei strain GG reverses increased intestinal permeability induced by cow milk in suckling rats. Gastroenterology 105(6):1643–1650PubMedGoogle Scholar
  10. 10.
    Salminen S, Isolauri E, Salminen E (1996) Clinical uses of probiotics for stabilizing the gut mucosal barrier: successful strains and future challenges. Antonie Van Leeuwenhoek 70(2–4):347–358PubMedCrossRefGoogle Scholar
  11. 11.
    Lu L, Walker WA (2001) Pathologic and physiologic interactions of bacteria with the gastrointestinal epithelium. Am J Clin Nutr 73(6):1124S–1130SPubMedGoogle Scholar
  12. 12.
    Resta-Lenert S, Barrett KE (2003) Live probiotics protect intestinal epithelial cells from the effects of infection with enteroinvasive Escherichia coli (EIEC). Gut 52(7):988–997PubMedCrossRefGoogle Scholar
  13. 13.
    Montalto M, Maggiano N, Ricci R, Curigliano V, Santoro L, Di Nicuolo F, Vecchio FM, Gasbarrini A, Gasbarrini G (2004) Lactobacillus acidophilus protects tight junctions from aspirin damage in HT-29 cells. Digestion 69(4):225–228. doi: 10.1159/000079152 PubMedCrossRefGoogle Scholar
  14. 14.
    Gaudier E, Michel C, Segain JP, Cherbut C, Hoebler C (2005) The VSL# 3 probiotic mixture modifies microflora but does not heal chronic dextran-sodium sulfate-induced colitis or reinforce the mucus barrier in mice. J Nutr 135(12):2753–2761PubMedGoogle Scholar
  15. 15.
    Mennigen R, Nolte K, Rijcken E, Utech M, Loeffler B, Senninger N, Bruewer M (2009) Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction protein expression and preventing apoptosis in a murine model of colitis. Am J Physiol Gastrointest Liver Physiol 296(5):G1140–G1149. doi: 10.1152/ajpgi.90534.2008 PubMedCrossRefGoogle Scholar
  16. 16.
    Kim HJ, Camilleri M, McKinzie S, Lempke MB, Burton DD, Thomforde GM, Zinsmeister AR (2003) A randomized controlled trial of a probiotic, VSL#3, on gut transit and symptoms in diarrhoea-predominant irritable bowel syndrome. Aliment Pharmacol Ther 17(7):895–904PubMedCrossRefGoogle Scholar
  17. 17.
    Kim HJ, Vazquez Roque MI, Camilleri M, Stephens D, Burton DD, Baxter K, Thomforde G, Zinsmeister AR (2005) A randomized controlled trial of a probiotic combination VSL# 3 and placebo in irritable bowel syndrome with bloating. Neurogastroenterol Motil 17(5):687–696. doi: 10.1111/j.1365-2982.2005.00695.x PubMedCrossRefGoogle Scholar
  18. 18.
    Guandalini S, Magazzu G, Chiaro A, La Balestra V, Di Nardo G, Gopalan S, Sibal A, Romano C, Canani RB, Lionetti P, Setty M (2010) VSL#3 improves symptoms in children with irritable bowel syndrome: a multicenter, randomized, placebo-controlled, double-blind, crossover study. J Pediatr Gastroenterol Nutr 51(1):24–30. doi: 10.1097/MPG.0b013e3181ca4d95 PubMedCrossRefGoogle Scholar
  19. 19.
    Camilleri M, Heading RC, Thompson WG (2002) Clinical perspectives, mechanisms, diagnosis and management of irritable bowel syndrome. Aliment Pharmacol Ther 16(8):1407–1430PubMedCrossRefGoogle Scholar
  20. 20.
    Alaradi O, Barkin JS (2002) Irritable bowel syndrome: update on pathogenesis and management. Med Princ Pract 11(1):2–17PubMedCrossRefGoogle Scholar
  21. 21.
    Frieling T (2006) Functional and inflammatory bowel disorders. Med Klin (Munich) 101(Suppl 1):139–142Google Scholar
  22. 22.
    Crentsil V (2005) Will corticosteroids and other anti-inflammatory agents be effective for diarrhea-predominant irritable bowel syndrome? Med Hypotheses 65(1):97–102. doi: 10.1016/j.mehy.2004.07.042 PubMedCrossRefGoogle Scholar
  23. 23.
    Kuiken SD, Klooker TK, Tytgat GN, Lei A, Boeckxstaens GE (2006) Possible role of nitric oxide in visceral hypersensitivity in patients with irritable bowel syndrome. Neurogastroenterol Motil 18(2):115–122. doi: 10.1111/j.1365-2982.2005.00731.x PubMedCrossRefGoogle Scholar
  24. 24.
    Delvaux M (2002) Role of visceral sensitivity in the pathophysiology of irritable bowel syndrome. Gut 51(Suppl 1):i67–i71PubMedCrossRefGoogle Scholar
  25. 25.
    Coutinho SV, Urban MO, Gebhart GF (1998) Role of glutamate receptors and nitric oxide in the rostral ventromedial medulla in visceral hyperalgesia. Pain 78(1):59–69PubMedCrossRefGoogle Scholar
  26. 26.
    Coutinho SV, Gebhart GF (1999) A role for spinal nitric oxide in mediating visceral hyperalgesia in the rat. Gastroenterology 116(6):1399–1408PubMedCrossRefGoogle Scholar
  27. 27.
    Barbara G (2006) Mucosal barrier defects in irritable bowel syndrome. Who left the door open? Am J Gastroenterol 101(6):1295–1298. doi: 10.1111/j.1572-0241.2006.00667.x PubMedCrossRefGoogle Scholar
  28. 28.
    Barbara G, Cremon C, Pallotti F, De Giorgio R, Stanghellini V, Corinaldesi R (2009) Postinfectious irritable bowel syndrome. J Pediatr Gastroenterol Nutr 48(Suppl 2):S95–S97. doi: 10.1097/MPG.0b013e3181a15e2e PubMedCrossRefGoogle Scholar
  29. 29.
    Fanning AS, Jameson BJ, Jesaitis LA, Anderson JM (1998) The tight junction protein ZO-1 establishes a link between the transmembrane protein occludin and the actin cytoskeleton. J Biol Chem 273(45):29745–29753PubMedCrossRefGoogle Scholar
  30. 30.
    Gonzalez-Mariscal L, Betanzos A, Nava P, Jaramillo BE (2003) Tight junction proteins. Prog Biophys Mol Biol 81(1):1–44PubMedCrossRefGoogle Scholar
  31. 31.
    Brookes SJ (2001) Classes of enteric nerve cells in the guinea-pig small intestine. Anat Rec 262(1):58–70. doi: 10.1002/1097-0185(20010101)262:1<58:AID-AR1011>3.0.CO;2-V PubMedCrossRefGoogle Scholar
  32. 32.
    La JH, Kim TW, Sung TS, Kang JW, Kim HJ, Yang IS (2003) Visceral hypersensitivity and altered colonic motility after subsidence of inflammation in a rat model of colitis. World J Gastroenterol 9(12):2791–2795PubMedGoogle Scholar
  33. 33.
    Al-Chaer ED, Kawasaki M, Pasricha PJ (2000) A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development. Gastroenterology 119(5):1276–1285PubMedCrossRefGoogle Scholar
  34. 34.
    Kitajima S, Takuma S, Morimoto M (1999) Changes in colonic mucosal permeability in mouse colitis induced with dextran sulfate sodium. Exp Anim 48(3):137–143PubMedCrossRefGoogle Scholar
  35. 35.
    Ukena SN, Singh A, Dringenberg U, Engelhardt R, Seidler U, Hansen W, Bleich A, Bruder D, Franzke A, Rogler G, Suerbaum S, Buer J, Gunzer F, Westendorf AM (2007) Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by enhancing mucosal integrity. PLoS One 2(12):e1308. doi: 10.1371/journal.pone.0001308 PubMedCrossRefGoogle Scholar
  36. 36.
    Kreiss C, Toegel S, Bauer AJ (2004) Alpha2-adrenergic regulation of NO production alters postoperative intestinal smooth muscle dysfunction in rodents. Am J Physiol Gastrointest Liver Physiol 287(3):G658–G666. doi: 10.1152/ajpgi.00526.2003 PubMedCrossRefGoogle Scholar
  37. 37.
    Hierholzer C, Kalff JC, Billiar TR, Bauer AJ, Tweardy DJ, Harbrecht BG (2004) Induced nitric oxide promotes intestinal inflammation following hemorrhagic shock. Am J Physiol Gastrointest Liver Physiol 286(2):G225–G233. doi: 10.1152/ajpgi.00447.2002 PubMedCrossRefGoogle Scholar
  38. 38.
    Garthwaite J, Charles SL, Chess-Williams R (1988) Endothelium-derived relaxing factor release on activation of NMDA receptors suggests role as intercellular messenger in the brain. Nature 336(6197):385–388. doi: 10.1038/336385a0 PubMedCrossRefGoogle Scholar
  39. 39.
    Fritz E, Hammer J, Schmidt B, Eherer AJ, Hammer HF (2003) Stimulation of the nitric oxide-guanosine 3′,5′-cyclic monophosphate pathway by sildenafil: effect on rectal muscle tone, distensibility, and perception in health and in irritable bowel syndrome. Am J Gastroenterol 98(10):2253–2260. doi: 10.1111/j.1572-0241.2003.07661.x PubMedGoogle Scholar
  40. 40.
    Kuiken SD, Vergeer M, Heisterkamp SH, Tytgat GN, Boeckxstaens GE (2002) Role of nitric oxide in gastric motor and sensory functions in healthy subjects. Gut 51(2):212–218PubMedCrossRefGoogle Scholar
  41. 41.
    Rodella L, Rezzani R, Agostini C, Bianchi R (1998) Induction of NADPH-diaphorase activity in the rat periaqueductal gray matter after nociceptive visceral stimulation. Brain Res 793(1–2):333–336PubMedCrossRefGoogle Scholar
  42. 42.
    Vizzard MA, Erdman SL, de Groat WC (1995) Increased expression of neuronal nitric oxide synthase (NOS) in visceral neurons after nerve injury. J Neurosci 15(5 Pt 2):4033–4045PubMedGoogle Scholar
  43. 43.
    Bonaz B (2003) Visceral sensitivity perturbation integration in the brain-gut axis in functional digestive disorders. J Physiol Pharmacol 54(Suppl 4):27–42PubMedGoogle Scholar
  44. 44.
    Fooks LJ, Gibson GR (2002) Probiotics as modulators of the gut flora. Br J Nutr 88(Suppl 1):S39–S49. doi: 10.1079/BJN2002628 PubMedCrossRefGoogle Scholar
  45. 45.
    Verdu EF, Bercik P, Collins SM (2009) Effect of probiotics on gastrointestinal function: evidence from animal models. Therap Adv Gastroenterol 2(4):31–35. doi: 10.1177/1756283X09337645 PubMedCrossRefGoogle Scholar
  46. 46.
    Matsuo K, Zhang X, Ono Y, Nagatomi R (2009) Acute stress-induced colonic tissue HSP70 expression requires commensal bacterial components and intrinsic glucocorticoid. Brain Behav Immun 23(1):108–115. doi: 10.1016/j.bbi.2008.07.013 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Cong Dai
    • 1
  • Stefano Guandalini
    • 2
  • De-Hui Zhao
    • 1
  • Min Jiang
    • 1
  1. 1.Department of GastroenterologyFirst Affiliated Hospital, China Medical UniversityShenyang CityChina
  2. 2.Section of GastroenterologyCeliac Disease Center, University of Chicago Comer Children’s HospitalChicagoUSA

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