This is the third annual report of an international research network studying the cellular impact of laryngopharyngeal reflux (LPR) on laryngeal epithelium. The objective of this study was to investigate the presence of E-cadherin (epithelial cadherin; the intercellular junctional complex protein) in relation to the presence of (intracellular) pepsin and carbonic anhydrase isoenzyme III (CAIII).
Methods:
Fifty-four laryngeal biopsy specimens from 18 LPR patients were studied by immunohistochemistry and Western blotting for pepsin, E-cadherin, and CAIII. These data were compared to those from normal control subjects analyzed in another research study.
Results:
Intracellular pepsin was detected in LPR patients, but not in controls. E-cadherin expression was reduced in patients with LPR. Carbonic anhydrase III expression was not found in the vocal fold or in the majority of samples taken from the ventricle of LPR patients and was inversely associated with E-cadherin membranous expression.
Conclusions:
The findings of depleted E-cadherin and CAIII and the presence of pepsin appear to correlate with LPR. The reduced protective response indicated by the reduced expression of CAIII may play an important role in the disruption of the intercellular barrier associated with the down-regulation of E-cadherin.
KoufmanJA. Laryngopharyngeal reflux is different from classic gastroesophageal reflux disease. Ear Nose Throat J2002;81(suppl 2):7–9.
2.
OrlandoRC. Pathogenesis of gastroesophageal reflux disease. Am J Med Sci2003;326:274–8.
3.
HicksDMOursTMAbelsonTIVaeziMFRichterJE. The prevalence of hypopharynx findings associated with gastroesophageal reflux in normal volunteers. J Voice2002;16:564–79.
4.
LanasARoyoYOrtegoJMolinaMSainzR. Experimental esophagitis induced by acid and pepsin in rabbits mimicking human reflux esophagitis. Gastroenterolology1999;116:97–107.
LanasAISousaFLOrtegoJ. Aspirin renders the oesophageal mucosa more permeable to acid and pepsin. Eur J Gastroenterol Hepatol1995;7:1065–72.
7.
SchweitzerEJBassBLJohnsonLFHarmonJW. Sucralfate prevents experimental peptic esophagitis in rabbits. Gastroenterology1985;88:611–9.
8.
TayHPChaparalaRCHarmonJW. Bismuth subsalicylate reduces peptic injury of the oesophagus in rabbits. Gut1990;31:11–6.
9.
HopwoodD. Oesophageal damage and defence in reflux oesophagitis: Pathophysiological and cell biological mechanisms. Prog Histochem Cytochem1997;32:1–42.
10.
OrlandoRC. Gastroesophageal reflux disease: Offensive factors and tissue resistance. New York, NY: Dekker, 2000.
11.
GoldsteinJLWatkinsJLGreagerJALaydenTJ. The esophageal mucosal resistance: Structure and function of an unique gastrointestinal epithelial barrier. J Lab Clin Med1994;123:653–9.
12.
TobeyNACarsonJLAlkiekRAOrlandoRC. Dilated intercellular spaces: A morphological feature of acid reflux–damaged human esophageal epithelium. Gastroenterology1996;111:1200–5.
13.
OrlandoRC. Esophageal epithelial defenses against acid injury. Am J Gastroenterol1994;89(suppl):S48–S52.
14.
JohnstonNBulmerDGillGA. Cell biology of laryngeal epithelial defenses in health and disease: Further studies. Ann Otol Rhinol Laryngol2003;112:481–91.
15.
TobeyNAHosseiniSSCaymaz-BorCWyattHROrlandoGSOrlandoRC. The role of pepsin in acid injury to esophageal epithelium. Am J Gastroenterol2001;96:3062–70.
16.
KoufmanJA. The otolaryngologic manifestations of gastroesophageal reflux disease (GERD): A clinical investigation of 225 patients using ambulatory 24-hour pH monitoring and an experimental investigation of the role of acid and pepsin in the development of laryngeal injury. Laryngoscope1991;101(suppl 53).
17.
PostmaGNTomekMSBelafskyPCKoufmanJA. Esophageal motor function in laryngopharyngeal reflux is superior to that in classic gastroesophageal reflux disease. Ann Otol Rhinol Laryngol2001;110:1114–6.
18.
BellNJHuntRH. Role of gastric acid suppression in the treatment of gastro-oesophageal reflux disease. Gut1992;33:118–24.
19.
BremnerRMCrookesPFDeMeesterTRPetersJHSteinHJ. Concentration of refluxed acid and esophageal mucosal injury. Am J Surg1992;164:522–7.
20.
FarrowDCVaughanTLSweeneyC. Gastroesophageal reflux disease, use of H2 receptor antagonists, and risk of esophageal and gastric cancer. Cancer Causes Control2000;11:231–8.
Ismail-BeigiFHortonPFPopeCEII. Histological consequences of gastroesophageal reflux in man. Gastroenterology1970;58:163–74.
23.
TakeichiM. Cadherin cell adhesion receptors as a morphogenetic regulator. Science1991;251:1451–5.
24.
TakeichiM. Morphogenetic roles of classic cadherins. Curr Opin Cell Biol1995;7:619–27.
25.
SmithMEFPignatelliM. The molecular histology of neoplasia: The role of the cadherin/catenin complex. Histopathology1997;31:107–11.
26.
GumbinerBM. Regulation of cadherin adhesive activity. J Cell Biol2000;148:399–404.
27.
GumbinerBM. Cell adhesion: The molecular basis of tissue architecture and morphogenesis. Cell1996;84:345–57.
28.
SaloJALehtoVPKivilaaksoE. Morphological alterations in experimental esophagitis. Light microscopic and scanning and transmission electron microscopic study. Dig Dis Sci1983;28:440–8.
29.
ShimonoMClementiF. Intercellular junctions of oral epithelium. II. Ultrastructural changes in rat buccal epithelium induced by trypsin digestion. J Ultrastruct Res1977;59:101–12.
30.
AxfordSESharpNRossPE. Cell biology of laryngeal epithelial defenses in health and disease: Preliminary studies. Ann Otol Rhinol Laryngol2001;110:1099–108.
31.
TashianRE. The carbonic anhydrases: Widening perspectives on their evolution, expression and function. Bioessays1989;10:186–92.
32.
SlyWSHuPY. Human carbonic anhydrases and carbonic anhydrase deficiencies. Annu Rev Biochem1995;64:375–401.
33.
ParkkilaSParkkilaAK. Carbonic anhydrase in the alimentary tract. Roles of the different isozymes and salivary factors in the maintenance of optimal conditions in the gastrointestinal canal. Scand J Gastroenterol1996;31:305–17.
34.
ChristieKNThomsonCXueLLucocqJMHopwoodD. Carbonic anhydrase isoenzymes I, II, III, and IV are present in human esophageal epithelium. J Histochem Cytochem1997;45:35–40.
35.
TobeyNAPowellDWSchreinerVJOrlandoRC. Serosal bicarbonate protects against acid injury to rabbit esophagus. Gastroenterology1989;96:1466–77.
36.
HelmJFDoddsWJHoganWJSoergelKHEgideMSWoodCM. Acid neutralizing capacity of human saliva. Gastroenterology1982;83:69–74.
37.
BelafskyPCPostmaGNKoufmanJA. Validity and reliability of the reflux symptom index (RSI). J Voice2002;16:274–7.
38.
BelafskyPCPostmaGNKoufmanJA. The validity and reliability of the reflux finding score (RFS). Laryngoscope2001;111:1313–7.
39.
JohnstonNKnightJDettmarPWLivelyMOKoufmanJ. Pepsin and carbonic anhydrase isoenzyme III as diagnostic markers for laryngopharyngeal reflux disease. Laryngoscope2004;114:2129–34.
