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Stress Increases Susceptibility to Oxidative/Nitrosative Mucosal Damage in an Experimental Model of Colitis in Rats

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Abstract

Inflammatory bowel diseases (IBDs) are multifactorial processes. Clinical and animal studies indicate that emotional stress may contribute to the onset and progress of these diseases. On the other hand, enhanced free radical production in mucosal cells has been also implicated in the pathogenesis of IBD. Using an experimental model of colitis induced by intrarectal instillation of 2,4,6-trinitrobenzene sulfonic acid (TNBS) plus ethanol (vehicle), we sought to determine whether prior exposure to immobilization stress modifies the susceptibility to oxidative damage in colonic mucosa. Several groups of Wistar rats were used: control (C) and stressed (by immobilization of 6 hr every day during 10 days; S) groups and rats receiving a colitis-inducing dose of TNBS on day 5 (30 mg; TNBS30) and a noninflammatory dose of TNBS on day 5 (5 mg; TNBS5) with or without stress (prior exposure, days 0-5, and after, days 5-10). At the 10th day, colonic tissue was dissected and processed for biochemical studies. TNBS30 led to body weight loss, macroscopic colonic ulceration, and inflammation (determined by histological parameters and myeloperoxidase [MPO] activity) and to an increase in inducible nitric oxide synthase (NOS-2) activity and expression. TNBS5-instilled animals' body weight and biochemical inflammatory parameters were not significantly different from those in control animals. Interestingly, while stress did not modify body weight, macroscopic aspect of the mucosa, or NOS activity in animals receiving TNBS30, immobilization increased body weight loss, MPO levels, and malondialdehyde (MDA; an indicator of lipid peroxidation) levels after TNBS5. On the other hand, stress increased NOS-2 activity and immunohistochemical expression after instillation of TNBS5. Moreover, constitutive, Ca2+-dependent NOS activity decreased in stressed animals instilled with TNBS5 compared with nonstressed animals receiving TNBS5 (−28.5 ± 6.6%; P < 0.05). These findings indicate that previous exposure to stressful stimuli is a factor in susceptibility to oxidative damage in experimental colitis and support a possible protective effect of treatment of stress before and during the development of inflammation in the colon.

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REFERENCES

  1. Sullivan AJ: Psychogenic factors in ulcerative colitis. Am J Dig Dis Nutr 2:651-656, 1936

    Google Scholar 

  2. Levenstein S, Prantera C, Varvo V, Scribano ML, Berto E, Andreoli A, Luzi C: Psychological stress and disease activity in ulcerative colitis: a ultidimensional cross-sectional study. Am J Gastroenterol 89:1219-1225, 1994

    PubMed  Google Scholar 

  3. Levenstein S: Stress and ulcerative colitis: Convincing the doubting Thomases. Am J Gastroenterol 98:2112-2115, 2003

    PubMed  Google Scholar 

  4. Farrell RJ, Peppercorn MA: Ulcerative colitis. Lancet 359:331-340, 2002

    Article  PubMed  Google Scholar 

  5. Bitton A, Sewitch MJ, Peppercorn MA, deB Edwardes MD, Shah S, Ransil B, Locke SE: Psychosocial determinants of relapse in ulcerative colitis:Alongitudinal study. AmJ Gastroenterol 98:2203-2208, 2003

    Google Scholar 

  6. Hart A, Kamm MA: Review article: Mechanisms of initiation and perpetuation of gut inflammation by stress. Aliment Pharmacol Ther 16:2017-2028, 2002

    Article  PubMed  Google Scholar 

  7. Qu XW, Wang H, Rozenfeld RA, Huang W, Hsueh W: Type I nitric oxide synthase is the predominant NOS in rat small intestine. Regulation by platelet-activating factor. Biochim Biophys Acta 1451:211-277, 1999

    Article  PubMed  Google Scholar 

  8. Wilson LM, Baldwin AL: Environmental stress causes mast cell degranulation, endothelial and epithelialchanges, and edema in the rat intestinal mucosa. Microcirculation 6:189-198, 1999

    Article  PubMed  Google Scholar 

  9. Soderholm JD, Yates DA, Gareau MG, Yang PC, MacQueen G, Perdue MH: Neonatal maternal separation predisposes adult rats to colonic barrier dysfunction in response to mild stress. Am J Physiol Gastrointest Liver Physiol 283:G1257-G1263, 2002

    PubMed  Google Scholar 

  10. Soderholm JD, Yang PC, Ceponis P, Vohra A, Riddell R, Sherman PM, Perdue MH: Chronic stress induces mast celldependent bacterial adherence and initiates mucosal inflammation in rat intestine. Gastroenterology 123:1099-1108, 2002

    Article  PubMed  Google Scholar 

  11. Calz´a L, Giardino L, Ceccatelli S: NOS mRNA in the paraventricular nucleus of young and old rats after immobilization stress. Neuroreport 4:627-630, 1993

    PubMed  Google Scholar 

  12. Mizock BA: Alterations in carbohydrate metabolism during stress: A review of the literature. Am J Med 98:75-84, 1995

    Article  PubMed  Google Scholar 

  13. Liu J, Wang X, Shigenaga MK, Yeo HC, Mori A, Ames BS: Immobilization stress causes oxidative damage to lipid, protein and DNA in the brain of rats. FASEB J 10:1532-1538, 1996

    PubMed  Google Scholar 

  14. Leza JC, Salas E, Sawicki G, Russell JC, Radomski MW: The effects of stress on homeostasis in JCR-LA-cp rats: The role of nitric oxide J Pharmacol Exp Ther 286:1397-1403, 1998

    PubMed  Google Scholar 

  15. Olivenza R, Moro MA, Lizasoain I, Lorenzo P, Fern´andez AP, Rodrigo J, Bosc´a L, Leza JC: Chronic stress induces the expression of inducible nitric oxide synthase in rat brain cortex. J Neurochem 74:785-791, 2000

    Article  PubMed  Google Scholar 

  16. Boughton-Smith NK, Evans SM, Laszlo F, Whittle BJR, Moncada S: The induction of nitric oxide synthase and intestinal vascular permeability by endotoxin in the rat. Br J Pharmacol 110:1189-1195, 1993

    PubMed  Google Scholar 

  17. Middleton SJ, Sorthouse M, Hunter JD: Increased nitric oxide synthesis in ulcerative colitis. Lancet 341:465-466, 1993

    Article  PubMed  Google Scholar 

  18. Rachmilewitz D, Stamler JS, Bachwich D, Karmeli F, Ackerman Z, Podolsky DK: Enhanced colonic nitric oxide generation and nitric oxide synthase activity in ulcerative colitis and Crohn's disease. Gut 36:718-723, 1995

    PubMed  Google Scholar 

  19. Kimura H, Hokari R, Miura S, Shigematsu T, Hirokawa M, Akiba Y, Kurose I, Higuchi H, Fujimori H, Tsuzuki Y, Serizawa H, Ishii H: Increased expression of an inducible isoform of nitric oxide synthase and the formation of peroxynitrite in colonic mucosa of patients with active ulcerative colitis. Gut 42:180-187, 1998

    PubMed  Google Scholar 

  20. Col´on AL, Menchén LA, Lizasoain I, Leza JC, Menchén PL, Gonz´alez-Lara V, Moro MA, Lorenzo P: Inducible nitric oxide synthase activity is expressed not only in inflamed but also in normal colonic mucosa in patients with ulcerative colitis. A potential prognostic marker. Am J Gastroenterol 95:1371-1373, 2000

    PubMed  Google Scholar 

  21. Miller MJS, Sadowska-Krowicka H, Chotinaruemol S, Kakkis JL, Clark DA: Amelioration of chronic ileitis by nitric oxide synthase inhibition. J Pharmacol Exp Ther 264:11-16, 1993

    PubMed  Google Scholar 

  22. Miller MJS, Zhang XJ, Sadowska-Krowicka H, Chotinaruemol S, Meintyre JA, Clark DA, Bustamante SA: Nitric oxide release in response to gut injury. Scand J Gastroenterol 28:1490-154, 1993

    Google Scholar 

  23. Miller MJS, Thompson JH, Zhang XJ, Kakkis JL, Munshi UK, Sandoval M, Rossi JL, Eloby-Childress S, Beckman JS, Ye Yao Z, Rodi CP, Manning PT, Curie MG, Clark DA: Role of inducible nitric oxide synthase expression and peroxynitrite formation in guinea pig ileitis. Gastroenterology 109:1475-1483, 1995

    PubMed  Google Scholar 

  24. Boughton-Smith NK, Evans SM, Whittle BJR: Characterisation of nitric oxide synthase activity in the rat colonic mucosa and muscle after endotoxin and in a model of colitis. Agents Actions 41:C223-C225, 1994

    Google Scholar 

  25. Seo HG, Takata I, Nakamura M, Tatsumi H, Suzuki K, Fujii J, Taniguchi N: Induction of nitric oxide synthase and concomitant suppression of superoxide dismutases in experimental colitis in rats. Arch Biochem Biophys 324:41-47, 1995

    Article  PubMed  Google Scholar 

  26. Palmer RM, Bridge L, Foxwell NA, Moncada S: The role of nitric oxide in endothelial cell damage and its inhibition by glucocorticoids. Br J Pharmacol 105:11-12, 1992

    PubMed  Google Scholar 

  27. Tepperman BL, Vozzolo BL, Soper BD: Effect of neutropenia on gastric mucosal integrity and mucosal nitric oxidesynthesis in the rat. Dig Dis Sci 38:2056-2061, 1993

    PubMed  Google Scholar 

  28. Whittle BJR, Laszlo F, Evans SM, Moncada S: Induction of nitric oxide synthase and microvascular injury in the rat jejunum provoked by indometacin. Br J Pharmacol. 116:2286-2290, 1995

    PubMed  Google Scholar 

  29. Zingarelli B, Cuzzocrea S, Szabo C, Salzman A: Mercaptoethylguanidine, a combined inhibitor of nitric oxide synthase and peroxynitrite scavenger, reduces trinitrobenzene sulfonic acid-induced colonic damage in rats. J Pharmacol Exp Ther 287:1048-1055, 1998

    PubMed  Google Scholar 

  30. Menchén L, Col´on A, Moro MA, Leza JC, Lizasoain I, Menchén P, Alvarez E, Lorenzo P: N-(3-(Aminomethyl)benzyl)acetamidine, an inducible nitric oxide synthase inhibitor, decreases colonic inflammation induced by trinitrobenzene sulphonic acid in rats. Life Sci 69:479-491, 2001

    Article  PubMed  Google Scholar 

  31. Morris GP, Beck PL, Herridge MS, Depew WT, Szewczuk MR, Wallace JL: Hapten-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology 96:795-803, 1989

    PubMed  Google Scholar 

  32. Bell CJ, Gall DG, Wallace JL: Disruption of colonic electrolyte transport in experimental colitis. Am J Physiol 268(4, Pt 1):G622-G630, 1995

    PubMed  Google Scholar 

  33. Bradley PP, Priebat DA, Christensen RD: Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol 78:206-209, 1982

    Article  PubMed  Google Scholar 

  34. Salter M, Knowles RG, Moncada S:Widespread tissue distribution, species distribution and changes in activity of Ca2C-dependent and Ca2C-independent nitric oxide synthases. FEBS Lett 291:145-154, 1991

    Article  PubMed  Google Scholar 

  35. Rees DD, Cunha FQ, Assreuy J, Herman AG, Moncada S: Sequential induction of nitric oxide synthase by Corynebacterium parvum in different organs of the mouse. Br J Pharmacol 114:689-693, 1995

    PubMed  Google Scholar 

  36. Das NP, Ratty AK: Studies on the effects of the narcotic alkaloids, cocaine, morphine and codeine on nonenzymatic lipid peroxidation in rat brain mitochondria. Biochem Med Metab Biol 37: 256-264, 1987

    Google Scholar 

  37. Hill HD, Straka JG: Protein determination using bicinchoninic acid in the presence of sulfhydryl reagents. Anal Biochem 170:203-208, 1988

    PubMed  Google Scholar 

  38. Collins SM, McHugh K, Jacobson K, Khan I, Riddell R, Murase K, Weingarten HP: Previous inflammation alters the response of the rat colon to stress. Gastroenterology 111:1509-1515, 1996

    PubMed  Google Scholar 

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

    Article  PubMed  Google Scholar 

  40. Anton PA, Shanahan F: Neuroimmunomodulation in inflammatory bowel disease. How far from "bench" to "bedside"? Ann NY Acad Sci 840:723-734, 1998

    PubMed  Google Scholar 

  41. Milde AM, Murison R: A study of the effects of restraint stress on colitis induced by dextran sulphate sodium in singly housed rats. Integr Physiol Behav Sci 37:140-150, 2002

    PubMed  Google Scholar 

  42. Pfeiffer CJ, Qiu B, Lam SK: Reduction of colonic mucus by repeated short-term stress enhances experimental colitis in rats. J Physiol Paris 95:81-87, 2001

    Article  PubMed  Google Scholar 

  43. Qiu BS, Vallance BA, Blennerhassett PA, Collins SM: The role of CD4 C lymphocytes in the susceptibility of mice to stress-induced reactivation of experimental colitis. Nat Med 5:1178-1182, 1999

    Article  PubMed  Google Scholar 

  44. Stendahl O, Molin L, Lindroth M: Granulocyte-mediated release of histamine from mast cells. Effect of myeloperoxidase and its inhibition by antiinflammatory sulfone compounds. Int Arch Allergy Appl Immunol 70:277-284, 1983

    PubMed  Google Scholar 

  45. Grisham MB, Granger DN: Neutrophil-mediated mucosal injury. Role of reactive oxygen metabolites. Dig Dis Sci. 33(Suppl 3):6S-15S, 1988

    PubMed  Google Scholar 

  46. Cross RK, Wilson KT: Nitric oxide in inflammatory bowel disease. Inflamm Bowel Dis 9:179-189, 2003

    Article  PubMed  Google Scholar 

  47. McKenzie SJ, Baker MS, Buffinton GD, Doe WF: Evidence of oxidant-induced injury to epithelial cells during inflammatory bowel disease. J Clin Invest 98:136-141, 1996

    PubMed  Google Scholar 

  48. . Kruidenier L, Kuiper I, Van Duijn W, Mieremet-Ooms MA, van Hogezand RA, Lamers CB, Verspaget HW: Imbalanced secondary mucosal antioxidant response in inflammatory bowel disease. J Pathol 201:17-27, 2003

    Article  PubMed  Google Scholar 

  49. Lih-Brody L, Powell SR, Collier KP, Reddy GM, Cerchia R, Kahn E, Weissman GS, Katz S, loyd RA, McKinley MJ, Fisher SE, Mullin GE: Increased oxidative stress and decreased antioxidant defenses in mucosa of inflammatory bowel disease. Dig Dis Sci 41:2078-2086, 1996

    PubMed  Google Scholar 

  50. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA: Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc Nat Acad Sci USA 87:1620-1624, 1990

    PubMed  Google Scholar 

  51. . Esterbauer H, Schaur RJ, Zollner H: Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med 11:81-128, 1991

    Article  PubMed  Google Scholar 

  52. Mattson MP: Modification of ion homeostasis by lipid peroxidation: roles in neuronal degeneration and adaptive plasticity. Trends Neurosci 21:53-57, 1998

    Article  PubMed  Google Scholar 

  53. Madrigal JLM, Olivenza R, Moro MA, Lizasoain I, Lorenzo P, Rodrigo P, Leza JC: Glutathione depletion, lipid peroxidation and mitochondrial disfunction are induced by chronic stress in rat brain. Neuropsychopharmacology 24:420-429, 2001

    Article  PubMed  Google Scholar 

  54. D'Odorico A, Bortolan S, Cardin R, D'Inca' R, Martines D, Ferronato A, Sturniolo GC: Reduced plasma antioxidant concentrations and increased oxidative DNA damage in inflammatory bowel disease.Scand J Gastroenterol 36:1289-1294, 2001

    Article  PubMed  Google Scholar 

  55. Grisham MB, MacDermott RP, Deitch EA: Oxidant defense mechanisms in the human colon. Inflammation 14:669-680, 1990

    PubMed  Google Scholar 

  56. Wallace JL, Miller MJS: Nitric oxide in mucosal defense: A little goes a long away. Gastroenterology, 119:512-520, 2000

    PubMed  Google Scholar 

  57. Rolle U, Nemeth L, Puri P: Nitrergic innervation of the normal gut and in motility disorders of childhood. J Pediatr Surg 37:551-567, 2002

    Article  PubMed  Google Scholar 

  58. Percy WH, Burton MB, Rose K, Donovan V, Burakoff: In vitro changes in the properties or rabbit muscularis colonic mucosae in colitis. Gastroenterology 104:369-376, 1993

    PubMed  Google Scholar 

  59. Mizuta Y, Isomoto H, Takahashi T: Impaired nitrergic innervation in rat colitis induced by dextran sulfate sodium. Gastroenterology 118:714-723, 2000

    PubMed  Google Scholar 

  60. Depoortere I, Thijs T, Peeters TL: Generalized loss of inhibitory innervation reverses serotoninergic inhibition into excitation in a rabbit model of TNBS-colitis. Br J Pharmacol 135:2011-2019, 2002

    PubMed  Google Scholar 

  61. Lamas S, MichelT, BrennerBM, Marsden PA: Nitric oxide synthesis in endothelial cells: Evidence for a pathway inducible by TNF-alpha. Am J Physiol C634-C641, 1991

  62. Lelchuk R, Radomski MW, Martin JF, Moncada S: Constitutive and inducible nitric oxide synthases in human megakaryoblastic cells. J Pharmacol Exp Ther 262:12200-12204, 1992

    Google Scholar 

  63. De Alba J, C´ardenas A, Moro MA, Leza JC, Lorenzo P, Bosc´a L, Lizasoain I: Down-regulation of neuronal nitric oxide synthase by nitric oxide after oxygen-glucose deprivation in rat forebrain slices. J Neurochem 72:248-254, 1999

    Article  PubMed  Google Scholar 

  64. Huhmer AF, Gerber NC, de Montellano PR, Schoneich C: Peroxynitrite reduction of calmodulin stimulation of neuronal nitric oxide synthase. Chem Res Toxicol 9:484-491, 1996

    Article  PubMed  Google Scholar 

  65. Bandyopadhyay A, Chadker S, Rattan S: Regulation of inducible and neuronal nitric oxide synthase gene expression by interferongamma and VIP. Am J Physiol 272:C1790-C1797, 1997

    PubMed  Google Scholar 

  66. Gath I, Godtel-Armbrust U, Forsterman U: Expressional downregulation of neuronal-type NO synthase I in guinea pig skeletal muscle in response to bacterial lipopolysaccharide. FEBS Lett 410:319-323, 1997

    Article  PubMed  Google Scholar 

  67. Qu XW, Wang H, Rozenfeld RA, Huang W, Hsueh W: Type I nitric oxide synthase (NOS) is the predominant NOS in rat small intestine. Regulation by platelet-activating factor. Biochim Biophys Acta 1451:211-217, 1999

    Article  PubMed  Google Scholar 

  68. Kovacs KJ, Cai Y, Larson AA: Neuronal nitric oxide synthase mRNA is down-regulated, and constitutive NOS enzymatic activity decreased, in thoracic dorsal root ganglia and spinal cord of the rat by substance PN-terminal metabolite. Eur J Neurosci 14:577-584, 2001

    Article  PubMed  Google Scholar 

  69. Eliakim R, Karmeli F, Razin E, et al.: Role of platelet-activating factor in ulcerative colitis. Enhanced production during active disease and inhibition by sulphasalazine and prednisolone. Gastroenterology 95:1167-1172, 1988

    PubMed  Google Scholar 

  70. Bernstein CN, Robert ME, Eysselein VE: Rectal substance P concentrations are increased in ulcerative colitis but not in Crohn's disease. Am J Gastroenterol 88:908-913, 1993

    PubMed  Google Scholar 

  71. Menchén L, Col´on A, Madrigal JLM, Beltr´an L, Botella S, Lizasoain I, Leza JC, Moro MA, Menchén P, Cos E, Lorenzo P: Activity of inducible and neuronal nitric oxide synthases in colonic mucosa predicts progression of ulcerative colitis inflammatory extension. Am J Gastroenterol 99:1756-1764, 2004

    Article  PubMed  Google Scholar 

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  1. Departamento de Farmacología, Facultad de Medicina, Universidad Complutense Madrid, Madrid, 28040, Spain

    Arturo L. Colón, José L. M. Madrigal, Luis A. Menchén, María A. Moro, Ignacio Lizasoain, Pedro Lorenzo & Juan C. Leza

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  1. Arturo L. Colón
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  3. Luis A. Menchén
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Colón, A.L., Madrigal, J.L.M., Menchén, L.A. et al. Stress Increases Susceptibility to Oxidative/Nitrosative Mucosal Damage in an Experimental Model of Colitis in Rats. Dig Dis Sci 49, 1713–1721 (2004). https://doi.org/10.1023/B:DDAS.0000043391.64073.e4

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