Abstract
Recent investigation of the intestine following ischemia and reperfusion (I/R) has revealed that nitric oxide synthase (NOS) neurons are more strongly affected than other neuron types. This implies that NO originating from NOS neurons contributes to neuronal damage. However, there is also evidence of the neuroprotective effects of NO. In this study, we compared the effects of I/R on the intestines of neuronal NOS knockout (nNOS−/−) mice and wild-type mice. I/R caused histological damage to the mucosa and muscle and infiltration of neutrophils into the external muscle layers. Damage to the mucosa and muscle was more severe and greater infiltration by neutrophils occurred in the first 24 h in nNOS−/− mice. Immunohistochemistry for the contractile protein, α-smooth muscle actin, was used to evaluate muscle damage. Smooth muscle actin occurred in the majority of smooth muscle cells in the external musculature of normal mice but was absent from most cells and was reduced in the cytoplasm of other cells following I/R. The loss was greater in nNOS−/− mice. Basal contractile activity of the longitudinal muscle and contractile responses to nerve stimulation or a muscarinic agonist were reduced in regions subjected to I/R and the effects were greater in nNOS−/− mice. Reductions in responsiveness also occurred in regions of operated mice not subjected to I/R. This is attributed to post-operative ileus that is not significantly affected by knockout of nNOS. The results indicate that deleterious effects are greater in regions subjected to I/R in mice lacking nNOS compared with normal mice, implying that NO produced by nNOS has protective effects that outweigh any damaging effect of this free radical produced by enteric neurons.
Similar content being viewed by others
References
Bauer AJ, Boeckxstaens GE (2004) Mechanisms of postoperative ileus. Neurogastroenterol Motil 16:54–60
Beck PL, Xavier R, Wong J, Ezedi I, Mashimo H, Mizoguchi A, Mizoguchi E, Bhan AK, Podolsky DK (2004) Paradoxical roles of different nitric oxide synthase isoforms in colonic injury. Am J Physiol 286:G137–G147
Ferrer-Sueta G, Radi R (2009) Chemical biology of peroxynitrite: kinetics, diffusion, and radicals. ACS Chem Biol 4:161–177
Furness JB, Costa M (1979) Projections of intestinal neurons showing immunoreactivity for vasoactive intestinal polypeptide are consistent with these neurons being the enteric inhibitory neurons. Neurosci Lett 15:199–204
Furness JB, Kunze WAA, Clerc N (1999) Nutrient tasting and signaling mechanisms in the gut. II. The intestine as a sensory organ: neural, endocrine, and immune responses. Am J Physiol 277:G922–G928
Gabella G (1974) Special muscle cells and their innervation in the mammalian small intestine. Cell Tissue Res 153:63–77
Granger N, Korthuis RJ (1995) Physiologic mechanisms of postischemic tissue injury. Annu Rev Physiol 57:311–332
Grisham MB, Granger DN, Lefer DJ (1998) Modulation of leukocyte-endothelial interactions by reactive metabolites of oxygen and nitrogen: relevance to ischemic heart disease. Free Radic Biol Med 25:404–433
Hierholzer C, Kalff JC, Audolfsson G, Billiar TR, Tweardy DJ, Bauer AJ (1999) Molecular and functional contractile sequelae of rat intestinal ischemia/reperfusion injury. Transplantation 68:1244–1254
Hoffman RA, Zhang G, Nussler NC, Gleixner SL, Ford HR, Simmons RL, Watkins SC (1997) Constitutive expression of inducible nitric oxide synthase in the mouse ileal mucosa. Am J Physiol Gastrointest Liver Physiol 272:G383–G392
Huang PL, Dawson TM, Bredt DS, Snyder SH, Fishman MC (1993) Targeted disruption of the neuronal nitric oxide synthase gene. Cell 75:1273–1286
Kalff JC, Schraut WH, Simmons RL, Bauer AJ (1998a) Surgical manipulation of the gut elicits an intestinal muscularis inflammatory response resulting in postsurgical ileus. Ann Surg 228:652–663
Kalff JC, Schwarz NT, Walgenbach KJ, Schraut WH, Bauer AJ (1998b) Leukocytes of the intestinal muscularis: their phenotype and isolation. J Leukoc Biol 63:683–691
Kalff JC, Carlos TM, Schraut WH, Billiar TR, Simmons RL, Bauer AJ (1999) Surgically induced leukocytic infiltrates within the rat intestinal muscularis mediate postoperative ileus. Gastroenterology 117:378–387
Knowles CH, De Giorgio R, Kapur RP, Bruder E, Farrugia G, Geboes K, Gershon MD, Hutson J, Lindberg G, Martin JE, Meier-Ruge WA, Milla PJ, Smith VV, Vandervinden JM, Veress B, Wedel T (2009) Gastrointestinal neuromuscular pathology: guidelines for histological techniques and reporting on behalf of the Gastro 2009 International Working Group. Acta Neuropathol 118:271–301
Kobayashi Y (2010) The regulatory role of nitric oxide in proinflammatory cytokine expression during the induction and resolution of inflammation. J Leukoc Biol 88:1157–1162
Kosonen O, Kankaanranta H, Malo-Ranta U, Moilanen E (1999) Nitric oxide-releasing compounds inhibit neutrophil adhesion to endothelial cells. Eur J Pharmacol 382:111–117
Kubes P, McCafferty DM (2000) Nitric oxide and intestinal inflammation. Am J Med 109:150–158
Kubes P, Suzuki M, Granger DN (1991) Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA 88:4651–4655
Kurose I, Kubes P, Wolf R, Anderson DC, Paulson J, Miyasada M, Granger DN (1993) Inhibition of nitric oxide production. Mechanisms of vascular albumin leakage. Circ Res 73:164–171
Lipton P (1999) Ischemic cell death in brain neurons. Physiol Rev 79:1431–1568
Livingston EH, Passaro EP Jr (1990) Postoperative ileus. Dig Dis Sci 35:121–132
Love S (1999) Oxidative stress in brain ischemia. Brain Pathol 9:119–131
Lubbers T, Buurman W, Luyer M (2010) Controlling postoperative ileus by vagal activation. World J Gastroenterol 16:1683–1687
Luo CC, Chen HM, Chiu CH, Lin JN, Chen JC (2001) Effect of NG-nitro-L-arginine methyl ester on intestinal permeability following intestinal ischemia-reperfusion injury in a rat model. Biol Neonate 80:60–63
Mallick IH, Yang W, Winslet MC, Seifalian AM (2004) Ischemia—reperfusion injury of the intestine and protective strategies against injury. Dig Dis Sci 49:1359–1377
Massberg S, Messmer K (1998) The nature of ischemia/reperfusion injury. Transplant Proc 30:4217–4223
Masuda E, Kawano S, Nagano K, Tsuji S, Takei Y, Tsujii M, Oshita M, Michida T, Kobayashi I, Nakama A, Fusamoto H, Kamada T (1995) Endogenous nitric oxide modulates ethanol-induced gastric mucosal injury in rats. Gastroenterology 108:58–64
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
Miller MJS, Munshi UK, Sadowska Krowicka H, Kakkis JL, Zhang XJ, Eloby-Childress S, Clark DA (1994) Inhibition of calcium-dependent nitric oxide synthase causes ileitis and leukocytosis in guinea pigs. Dig Dis Sci 39:1185–1192
Moro MA, Cárdenas A, Hurtado O, Leza JC, Lizasoain I (2004) Role of nitric oxide after brain ischaemia. Cell Calcium 36:265–275
Owens GK, Kumar MS, Wamhoff BR (2004) Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev 84:767–801
Payne D, Kubes P (1993) Nitric oxide donors reduce the rise in reperfusion-induced intestinal mucosal permeability. Am J Physiol Gastrointest Liver Physiol 265:G189–G195
Pfeiffer CJ, Qiu BS (1995) Effects of chronic nitric oxide synthase inhibition on TNB-induced colitis in rats. J Pharm Pharmacol 47:827–832
Pique JM, Whittle BJR, Esplugues JV (1989) The vasodilator role of endogenous nitric oxide in the rat gastric microcirculation. Eur J Pharmacol 174:293–296
Pontell L, Sharma P, Rivera LR, Thacker M, Tan YH, Brock JA, Furness JB (2011) Damaging effects of ischemia/reperfusion on intestinal muscle. Cell Tissue Res 343:411–419
Qu X-W, Wang H, De Plaen IG, Rozenfeld RA, Hsueh W (2001) Neuronal nitric oxide synthase (NOS) regulates the expression of inducible NOS in rat small intestine via modulation of nuclear factor kappa B. FASEB J 15:439–446
Rivera LR, Thacker M, Castelucci P, Bron R, Furness JB (2009) The reactions of specific neuron types to intestinal ischemia in the guinea pig enteric nervous system. Acta Neuropathol 118:261–270
Rivera LR, Poole DP, Thacker M, Furness JB (2011a) The involvement of nitric oxide synthase neurons in enteric neuropathies. Neurogastroenterol Motil 23:980–988
Rivera LR, Thacker M, Pontell L, Cho H-J, Furness JB (2011b) Deleterious effects of intestinal ischemia/reperfusion injury in the mouse enteric nervous system are associated with protein nitrosylation. Cell Tissue Res 344:111–123
Taha MO, Miranda-Ferreira R, Fagundes AL, Fagundes DJ, Simões RS, Santos JM, Souza PDF, Júnior ISO, Marchini A, Gomes IT, Monteiro HP, Mendonça LO, Caricati-Neto A (2010) Effects of L-nitro-arginine methyl ester, an inhibitor of nitric oxide biosynthesis, on intestinal ischemia/reperfusion injury in rabbits. Transplant Proc 42:457–460
Togashi H, Sasaki M, Frohman E, Taira E, Ratan RR, Dawson TM, Dawson VL (1997) Neuronal (type I) nitric oxide synthase regulates nuclear factor κB activity and immunologic (type II) nitric oxide synthase expression. Proc Natl Acad Sci USA 94:2676–2680
Torihashi S, Horowitz B, Pollock JS, Ward SM, Xue C, Kobayashi S, Sanders KM (1996) Expression of nitric oxide synthase in mucosal cells of the canine colon. Histochem Cell Biol 105:33–41
Türler A, Kalff JC, Heeckt P, Abu-Elmagd KM, Schraut WH, Bond GJ, Moore BA, Brünagel G, Bauer AJ (2002) Molecular and functional observations on the donor intestinal muscularis during human small bowel transplantation. Gastroenterology 122:1886–1897
Woodruff TM, Thundyil J, Tang S-C, Sobey CG, Taylor SM, Arumugam TV (2011) Pathophysiology, treatment, and animal and cellular models of human ischemic stroke. Mol Neurodegener 6:1–19
Acknowledgments
We thank Associate Professor James Brock and Ms. Dorota Ferens for their helpful advice on the isometric tension recording experiments, Dr. Trung Nguyen for statistical advice and Mr. Gene Venables for his assistance in surgery and genotyping.
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by a grant from the National Health and Medical Research Council of Australia (grant 1006035).
Rights and permissions
About this article
Cite this article
Rivera, L.R., Pontell, L., Cho, HJ. et al. Knock out of neuronal nitric oxide synthase exacerbates intestinal ischemia/reperfusion injury in mice. Cell Tissue Res 349, 565–576 (2012). https://doi.org/10.1007/s00441-012-1451-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-012-1451-3