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Journal of Gastroenterology

Published in:

01-11-2016 | Original Article—Alimentary Tract

Serotonin disrupts esophageal mucosal integrity: an investigation using a stratified squamous epithelial model

Authors: Liping Wu, Tadayuki Oshima, Toshihiko Tomita, Yoshio Ohda, Hirokazu Fukui, Jiro Watari, Hiroto Miwa

Published in: Journal of Gastroenterology | Issue 11/2016

Abstract

Background

Serotonin regulates gastrointestinal function, and mast cells are a potential nonneuronal source of serotonin in the esophagus. Tight junction (TJ) proteins in the esophageal epithelium contribute to the barrier function, and the serotonin signaling pathway may contribute to epithelial leakage in gastroesophageal reflux disease. Therefore, the aim of this study was to investigate the role of serotonin on barrier function, TJ proteins, and related signaling pathways.

Methods

Normal primary human esophageal epithelial cells were cultured with use of an air–liquid interface system. Serotonin was added to the basolateral compartment, and transepithelial electrical resistance (TEER) was measured. The expression of TJ proteins and serotonin receptor 7 (5-HT₇) was assessed by Western blotting. The involvement of 5-HT₇ was assessed with use of an antagonist and an agonist. The underlying cellular signaling pathways were examined with use of specific blockers.

Results

Serotonin decreased TEER and reduced the expression of TJ proteins ZO-1, occludin, and claudin 1, but not claudin 4. A 5-HT₇ antagonist blocked the serotonin-induced decrease in TEER, and a 5-HT₇ agonist decreased TEER. Inhibition of p38 mitogen-activated protein kinase (MAPK) reduced the serotonin-induced decrease in TEER. Inhibition of p38 MAPK blocked the decrease of ZO-1 levels, whereas extracellular-signal-regulated kinase (ERK) inhibition blocked the decrease in occludin levels. Cell signaling pathway inhibitors had no effect on serotonin-induced alterations in claudin 1 and claudin 4 levels. Serotonin induced phosphorylation of p38 MAPK and ERK, and a 5-HT₇ antagonist partially blocked serotonin-induced phosphorylation of p38 MAPK but not that of ERK.

Conclusions

Serotonin disrupted esophageal squamous epithelial barrier function by modulating the levels of TJ proteins. Serotonin signaling pathways may mediate the pathogenesis of gastroesophageal reflux disease.

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Gershon MD. Review article: serotonin receptors and transporters—roles in normal and abnormal gastrointestinal motility. Aliment Pharmacol Ther. 2004;20(Suppl 7):3–14.CrossRefPubMed

Bertrand PP, Bertrand RL. Serotonin release and uptake in the gastrointestinal tract. Auton Neurosci. 2010;153:47–57.CrossRefPubMed

Hornung JP. The human raphe nuclei and the serotonergic system. J Chem Neuroanat. 2003;26:331–43.CrossRefPubMed

Watanabe H, Rose MT, Aso H. Role of peripheral serotonin in glucose and lipid metabolism. Curr Opin Lipidol. 2011;22:186–91.CrossRefPubMed

Tozzi A, Staiano A, Paparo F, et al. Characterization of the inflammatory infiltrate in peptic oesophagitis. Dig Liver Dis. 2001;33:452–8.CrossRefPubMed

Moons LM, Kusters JG, Bultman E, et al. Barrett’s oesophagus is characterized by a predominantly humoral inflammatory response. J Pathol. 2005;207:269–76.CrossRefPubMed

Heidmann DE, Metcalf MA, Kohen R, et al. Four 5-hydroxytryptamine₇ (5-HT₇) receptor isoforms in human and rat produced by alternative splicing: species differences due to altered intron-exon organization. J Neurochem. 1997;68:1372–81.CrossRefPubMed

Li HF, Liu JF, Zhang K, et al. Expression of serotonin receptors in human lower esophageal sphincter. Exp Ther Med. 2015;9:49–54.PubMed

Forster C. Tight junctions and the modulation of barrier function in disease. Histochem Cell Biol. 2008;130:55–70.CrossRefPubMedPubMedCentral

10.

Kushnir-Sukhov NM, Brown JM, Wu Y, et al. Human mast cells are capable of serotonin synthesis and release. J Allergy Clin Immunol. 2007;119:498–9.CrossRefPubMed

11.

Kushnir-Sukhov NM, Gilfillan AM, Coleman JW, et al. 5-Hydroxytryptamine induces mast cell adhesion and migration. J Immunol. 2006;177:6422–32.CrossRefPubMed

12.

Yang L, Cai H, Tou J, et al. The role of the 5-hydroxytryptamine pathway in reflux-induced esophageal mucosal injury in rats. World J Surg Oncol. 2012;10:219.CrossRefPubMedPubMedCentral

13.

Kimura Y, Shiozaki H, Hirao M, et al. Expression of occludin, tight-junction-associated protein, in human digestive tract. Am J Pathol. 1997;151:45–54.PubMedPubMedCentral

14.

Rendon-Huerta E, Valenzano MC, Mullin JM, et al. Comparison of three integral tight junction barrier proteins in Barrett’s epithelium versus normal esophageal epithelium. Am J Gastroenterol. 2003;98:1901–3.CrossRefPubMed

15.

Richter JE. Role of the gastric refluxate in gastroesophageal reflux disease: acid, weak acid and bile. Am J Med Sci. 2009;338:89–95.CrossRefPubMed

16.

Chen X, Oshima T, Tomita T, et al. Acidic bile salts modulate the squamous epithelial barrier function by modulating tight junction proteins. Am J Physiol Gastrointest Liver Physiol. 2011;301:G203–9.CrossRefPubMed

17.

Oshima T, Gedda K, Koseki J, et al. Establishment of esophageal-like non-keratinized stratified epithelium using normal human bronchial epithelial cells. Am J Physiol Cell Physiol. 2011;300:C1422–9.CrossRefPubMed

18.

Shan J, Oshima T, Muto T, et al. Epithelial-derived nuclear IL-33 aggravates inflammation in the pathogenesis of reflux esophagitis. J Gastroenterol. 2015;50:414–23.CrossRefPubMed

19.

Chen X, Oshima T, Shan J, et al. Bile salts disrupt human esophageal squamous epithelial barrier function by modulating tight junction proteins. Am J Physiol Gastrointest Liver Physiol. 2012;303:G199–208.CrossRefPubMed

20.

Oshima T, Koseki J, Chen X, et al. Acid modulates the squamous epithelial barrier function by modulating the localization of claudins in the superficial layers. Lab Investig. 2012;92:22–31.CrossRefPubMed

21.

Oshima T, Sasaki M, Kataoka H, et al. Wip1 protects hydrogen peroxide-induced colonic epithelial barrier dysfunction. Cell Mol Life Sci. 2007;64:3139–47.CrossRefPubMed

22.

Bueno L, Fioramonti J. Protease-activated receptor 2 and gut permeability: a review. Neurogastroenterol Motil. 2008;20:580–7.CrossRefPubMed

23.

Raymond JR, Mukhin YV, Gelasco A, et al. Multiplicity of mechanisms of serotonin receptor signal transduction. Pharmacol Ther. 2001;92:179–212.CrossRefPubMed

24.

Kim DY, Camilleri M. Serotonin: a mediator of the brain-gut connection. Am J Gastroenterol. 2000;95:2698–709.PubMed

25.

Pehlivanov N, Sarosiek I, Whitman R, et al. Effect of cisapride on nocturnal transient lower oesophageal sphincter relaxations and nocturnal gastro-oesophageal reflux in patients with oesophagitis: a double-blind, placebo-controlled study. Aliment Pharmacol Ther. 2002;16:743–7.CrossRefPubMed

26.

Willis S, Allescher HD, Stoschus B, et al. Double blind placebo controlled study on the effect of the nitric oxide donor molsidomin and the 5-HT3 antagonist ondansetron on human esophageal motility. Z Gastroenterol. 1994;32:632–6.PubMed

27.

Koutsoumbi P, Epanomeritakis E, Tsiaoussis J, et al. The effect of erythromycin on human esophageal motility is mediated by serotonin receptors. Am J Gastroenterol. 2000;95:3388–92.CrossRefPubMed

28.

Grossi L, Ciccaglione AF, Marzio L. Effect of the 5-HT1 agonist sumatriptan on oesophageal motor pattern in patients with ineffective oesophageal motility. Neurogastroenterol Motil. 2003;15:9–14.CrossRefPubMed

29.

Barnette MS, Grous M, Manning CD, et al. 5-Hydroxytryptamine (5-HT) and SK&F 103829 contract canine lower esophageal sphincter smooth muscle by stimulating 5-HT2 receptors. Receptor. 1992;2:155–67.PubMed

30.

Szczesniak MM, Fuentealba SE, Zhang T, et al. Modulation of esophageal afferent pathways by 5-HT3 receptor inhibition. Neurogastroenterol Motil. 2013;25:383–e293.CrossRefPubMed

31.

Liu H, Irving HR, Coupar IM. Expression patterns of 5-HT7 receptor isoforms in the rat digestive tract. Life Sci. 2001;69:2467–75.CrossRefPubMed

32.

Iceta R, Mesonero JE, Aramayona JJ, et al. Expression of 5-HT1A and 5-HT7 receptors in Caco-2 cells and their role in the regulation of serotonin transporter activity. J Physiol Pharmacol. 2009;60:157–64.PubMed

33.

Guseva D, Holst K, Kaune B, et al. Serotonin 5-HT7 receptor is critically involved in acute and chronic inflammation of the gastrointestinal tract. Inflamm Bowel Dis. 2014;20:1516–29.CrossRefPubMed

34.

Zou BC, Dong L, Wang Y, et al. Expression and role of 5-HT7 receptor in brain and intestine in rats with irritable bowel syndrome. Chin Med J (Engl). 2007;120:2069–74.

35.

Prins NH, Briejer MR, Van Bergen PJ, et al. Evidence for 5-HT7 receptors mediating relaxation of human colonic circular smooth muscle. Br J Pharmacol. 1999;128:849–52.CrossRefPubMedPubMedCentral

36.

Stull MA, Pai V, Vomachka AJ, et al. Mammary gland homeostasis employs serotonergic regulation of epithelial tight junctions. Proc Natl Acad Sci U S A. 2007;104:16708–13.CrossRefPubMedPubMedCentral

37.

Pai VP, Horseman ND. Biphasic regulation of mammary epithelial resistance by serotonin through activation of multiple pathways. J Biol Chem. 2008;283:30901–10.CrossRefPubMedPubMedCentral

38.

Witte AB, D’Amato M, Poulsen SS, et al. Duodenal epithelial transport in functional dyspepsia: role of serotonin. World J Gastrointest Pathophysiol. 2013;4:28–36.CrossRefPubMedPubMedCentral

39.

Piche T, Barbara G, Aubert P, et al. Impaired intestinal barrier integrity in the colon of patients with irritable bowel syndrome: involvement of soluble mediators. Gut. 2009;58:196–201.CrossRefPubMed

40.

Martinez C, Lobo B, Pigrau M, et al. Diarrhoea-predominant irritable bowel syndrome: an organic disorder with structural abnormalities in the jejunal epithelial barrier. Gut. 2013;62:1160–8.CrossRefPubMed

41.

Coeffier M, Gloro R, Boukhettala N, et al. Increased proteasome-mediated degradation of occludin in irritable bowel syndrome. Am J Gastroenterol. 2010;105:1181–8.CrossRefPubMed

42.

Bertiaux-Vandaele N, Youmba SB, Belmonte L, et al. The expression and the cellular distribution of the tight junction proteins are altered in irritable bowel syndrome patients with differences according to the disease subtype. Am J Gastroenterol. 2011;106:2165–73.CrossRefPubMed

43.

Zhang L, Liu G, Han X, et al. Inhibition of p38 MAPK activation attenuates esophageal mucosal damage in a chronic model of reflux esophagitis. Neurogastroenterol Motil. 2015;27:1648–56.CrossRefPubMed

44.

Costantini TW, Peterson CY, Kroll L, et al. Role of p38 MAPK in burn-induced intestinal barrier breakdown. J Surg Res. 2009;156:64–9.CrossRefPubMedPubMedCentral

45.

Wu HL, Gao X, Jiang ZD, et al. Attenuated expression of the tight junction proteins is involved in clopidogrel-induced gastric injury through p38 MAPK activation. Toxicology. 2013;304:41–8.CrossRefPubMed

46.

Ju Y, Wang T, Li Y, et al. Coxsackievirus B3 affects endothelial tight junctions: possible relationship to ZO-1 and F-actin, as well as p38 MAPK activity. Cell Biol Int. 2007;31:1207–13.CrossRefPubMed

47.

Kevil CG, Oshima T, Alexander B, et al. H₂O₂-mediated permeability: role of MAPK and occludin. Am J Physiol Cell Physiol. 2000;279:C21–30.PubMed

48.

Samak G, Aggarwal S, Rao RK. ERK is involved in EGF-mediated protection of tight junctions, but not adherens junctions, in acetaldehyde-treated Caco-2 cell monolayers. Am J Physiol Gastrointest Liver Physiol. 2011;301:G50–9.CrossRefPubMedPubMedCentral

49.

Kim EK, Choi EJ. Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta. 2010;1802:396–405.CrossRefPubMed

Title: Serotonin disrupts esophageal mucosal integrity: an investigation using a stratified squamous epithelial model
Authors: Liping Wu
Tadayuki Oshima
Toshihiko Tomita
Yoshio Ohda
Hirokazu Fukui
Jiro Watari
Hiroto Miwa
Publication date: 01-11-2016
Publisher: Springer Japan
Published in: Journal of Gastroenterology / Issue 11/2016
Print ISSN: 0944-1174
Electronic ISSN: 1435-5922
DOI: https://doi.org/10.1007/s00535-016-1195-z

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Results

Conclusions

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