Cyclooxygenase 1 and cyclooxygenase 2 expression is abnormally regulated in human nasal polyps

doi:10.1067/mai.2002.123534

Journal of Allergy and Clinical Immunology

Volume 109, Issue 5, May 2002, Pages 824-830

https://doi.org/10.1067/mai.2002.123534 Get rights and content

Abstract

Background: There is evidence that impairment of prostanoid metabolism might be involved in the pathogenesis of nasal polyps (NPs). Prostanoids are synthesized by 2 cyclooxygenase (Cox) enzymes, one constitutive (Cox-1) and another inducible (Cox-2). Objective: The aim of these studies was to investigate Cox-1 and Cox-2 regulation in NPs of aspirin-tolerant human patients compared with that seen in nasal mucosa (NM). Methods: Cultured explants from human NPs and healthy mucosa from patients undergoing polypectomy and corrective nasal surgery, respectively, were examined for Cox-1 and Cox-2 expression by means of semiquantitative competitive PCR and Western blotting. Results: Cox-1 mRNA was spontaneously upregulated in cultured NM but not in NPs. A spontaneous but delayed upregulation of Cox-2 mRNA was found in NPs (24 hours) compared with that seen in NM (6 hours). After cytokine stimulation (IFN-γ, IL-1β, and TNF-α), the induction of Cox-2 mRNA and protein was also faster in NM (1 hour) than in NPs (4 hours). Conclusion: These data showing an abnormal regulation of Cox-1 and Cox-2 in NPs from aspirin-tolerant patients reinforce the concept that prostanoid metabolism might be important in the pathogenesis of inflammatory nasal diseases and suggest a potential role for this alteration in the formation of NPs. (J Allergy Clin Immunol 2002;109:824-30.)

Section snippets

Materials

Penicillin-streptomycin, HEPES buffer, RPMI-1640 medium, Taq polymerase, Moloney murine leukemia virus reverse transcriptase, dithiothreitol, RNAse out, random primers, agarose, and PCR primers were from Life Technologies SA (Barcelona, Spain), and 24-well tissue culture clusters were from TPP (Barcelona, Spain). Amphotericin B was from Squibb (Esplugues de Llobregat, Spain). IL-1β, TNF-α, IFN-γ, and chloroform were from Sigma Chemical Co (Madrid, Spain). Isopropanol was from Panreac

Spontaneous regulation of Cox-1 and Cox-2 mRNA expression

Specimens from NM and NPs were collected and snap-frozen at −80°C at the time of the operation (0 hours) and after being in culture for 6, 24, 48, 72, 96, and 120 hours. Compared with hour 0 (NP, 0.24 ± 0.07; NM, 0.67 ± 0.15), spontaneous expression of Cox-1 mRNA in NM (n = 6), but not in NPs (n = 11), showed a time-dependent upregulation that was significant after 6 hours in culture (Fig 1).

. Spontaneous expression of genes encoding for Cox-1 and Cox-2 in NM (n = 7) and NPs (n = 11). *P < .05

Discussion

Our group previously reported low expression of Cox-2 in NPs of aspirin-sensitive asthmatic patients.¹⁸We now report that Cox-1 and Cox-2 are abnormally regulated in NPs obtained from aspirin-tolerant patients. The main findings of our study are as follows: (1) Cox-1 mRNA was spontaneously upregulated in cultured NM but not in NPs; (2) a spontaneous but delayed upregulation of Cox-2 mRNA was found in NPs compared with that in NM; and (3) the induction of Cox-2 mRNA and protein was also faster

References (41)

G Settipane et al.
Nasal polyps in asthma and rhinitis: a review of 6,037 patients
J Allergy Clin Immunol
(1977)
S Pinto et al.
Cyclooxygenase and lipoxygenase metabolite generation in nasal polyps
Prostaglandins Leukot Essent Fatty Acids
(1997)
D Auboeuf et al.
The use of reverse transcription-competitive polymerase chain reaction to investigate in vivo regulation of gene expression in small tissue samples
Anal Biochem
(1997)
A Ristimaki et al.
Induction of cyclooxygenase-2 by interleukin-1a. Evidence for post-transcriptional regulation
J Biol Chem
(1994)
W Smith et al.
Prostaglandin endoperoxide H synthases-1 and -2
Adv Immunol
(1996)
J Kitzler et al.
Analysis and quantification of splicing variants of TPA-inducible PGHS-1 mRNA in rat tracheal epithelial cells
Arch Biochem Biophys
(1995)
JL Wallace et al.
NSAID-induced gastric damage in rats: requirement for inhibition of both cyclooxygenase 1 and 2
Gastroenterology
(2000)
R Newton et al.
Superinduction of Cox-2 mRNA by cycloheximide and interleukin-1b involves increased transcription and correlates with increased NF-kB and JNK activation
FEBS Lett
(1997)
K Yamamoto et al.
Transcriptional roles of nuclear factor kappa B and nuclear factor-interleukin-6 in the tumor necrosis factor alpha-dependent induction of cyclooxygenase-2 in MC3T3-E1 cells
J Biol Chem
(1995)
P Economopoulos et al.
Glucocorticoids stimulate prostaglandin H synthase type-2 (PGHS-2) in the fibroblast cells in human amnion cultures
Mol Cell Endocrinol
(1996)

WL Smith et al.

Cyclooxygenases: structural, cellular, and molecular biology

Annu Rev Biochem

(2000)

J Raud et al.

Enhancement of acute allergic inflammation by indomethacin is reversed by prostaglandin E₂: apparent correlation with in vivo modulation of mediator release

Proc Natl Acad Sci U S A

(1998)

DW Gilroy et al.

Inducible cyclooxygenase may have antiinflammatory properties

Nat Med

(1999)

O Morteau et al.

Impaired mucosal defense to acute colonic injury in mice lacking cyclooxygenase-1 or cyclooxygenase-2

J Clin Invest

(2000)

RS Peebles et al.

Cyclooxygenase inhibition increases interleukin 5 and interleukin 13 production and airway responsiveness in allergic mice

Am J Respir Crit Care Med

(2000)

M. Hitchcock

Effect of inhibitors of prostaglandin synthesis and prostaglandins E₂ and F_2α on the immunologic release of mediators of inflammation from actively sensitized guinea-pig lung

J Pharmacol Exp Ther

(1978)

R. Djukanovic

Nasal polyps-a model of chronic respiratory mucosal inflammation

Clin Exp Allergy

(1995)

PL Larsen et al.

Origin and structure of nasal polyps

HW Baenkler et al.

Eicosanoids from biopsy of normal and polypous nasal mucosa

Rhinology

(1996)

SP Peters et al.

The pharmacological control of mediator release from human basophils and mast cells

Respiration

(1986)

Cited by (62)

Low E-prostanoid 2 receptor levels and deficient induction of the IL-1β/IL-1 type i receptor/COX-2 pathway: Vicious circle in patients with aspirin-exacerbated respiratory disease
2016, Journal of Allergy and Clinical Immunology
We hypothesized that the 2 reported alterations in aspirin-exacerbated respiratory disease (AERD), reduced expression/production of COX-2/prostaglandin (PG) E₂ and diminished expression of E-prostanoid (EP) 2 receptor, are closely linked.
We sought to determine the mechanisms involved in the altered regulation of the COX pathway in patients with AERD.
Fibroblasts were obtained from nasal mucosa; samples of control subjects (NM-C, n = 8) and from nasal polyps from patients with aspirin-exacerbated respiratory disease (NP-AERD, n = 8). Expression of the autocrine loop components regulating PGE₂ production and signaling, namely IL-1 type I receptor (IL-1RI), COX-2, microsomal prostaglandin E synthase 1 (mPGES-1), and EP receptors, was assessed at baseline and after stimulation with IL-1β, PGE₂, and specific EP receptor agonists.
Compared with NM-C fibroblasts, basal expression levels of IL-1RI and EP₂ receptor were lower in NP-AERD fibroblasts. IL-1β–induced IL-1RI, COX-2, and mPGES-1 expression levels were also lower in these cells. Levels of IL-1RI positively correlated with COX-2 and mPGES-1 expression in both NM-C and NP-AERD fibroblasts. Incubation with either exogenous PGE₂ or selective EP₂ agonist significantly increased expression of IL-1RI in NM-C fibroblasts and had hardly any effect on NP-AERD fibroblasts. Alterations in IL-1RI, COX-2, and mPGES-1 expression that were found in NP-AERD fibroblasts were corrected when EP₂ receptor expression was normalized by transfection of NP-AERD fibroblasts.
Altered expression of EP₂ in patients with AERD contributes to deficient induction of IL-1RI, reducing the capacity of IL-1β to increase COX-2 and mPGES-1 expression, which results in low PGE₂ production. This impairment in the generation of PGE₂ subsequently reduces its ability to induce IL-1RI.
Rhinosinusitis and Nasal Polyps in Aspirin-Exacerbated Respiratory Disease
2013, Immunology and Allergy Clinics of North America
Citation Excerpt :
Because Cox-2 expression increases in inflammatory processes, a concomitant alteration in the expression of Cox-2 would be expected to complement the changes in PGE2 levels detected in NP. In line with the results reported with PGE2, Cox-2 expression has been shown to be downregulated in NP of patients with and without aspirin hypersensitivity25,41–45 through a potential Nuclear Factor κ B–mediated downregulation.46 However, other studies failed to find any difference in the expression of Cox-2 in the NP of aspirin-intolerant, aspirin-tolerant, and control individuals.47
Current view on nasal polyps management in Samter's triad patients
2012, Otolaryngologia Polska
The nasal polyps associated with Samter's triad are often very extensive, difficult to treat, with great tendency to recurrence. In this paper the current opinion on nasal polyps management in aspirin triad patients was presented. Pathogenesis: Opinions on pathogenesis of these disease was remembered as well as its epidemiology. Diagnostic methods: The available diagnostic methods were presented. Treatment options: The available preservative treatment options was analyzed including aspirin desensitization. The role of surgical treatment, functional endoscopic sinus surgery was analyzed.
The effect of hypoxia and cycloxygenase inhibitors on nasal polyp derived fibroblasts
2011, American Journal of Otolaryngology - Head and Neck Medicine and Surgery
Citation Excerpt :
COX exists in two isoforms: COX-1, which is constitutively expressed in most tissues, and COX-2, which is inducible by a variety of agents such as inflammation and tissue damage. Altered regulation of both isoenzyems has been associated with the development of airway inflammation and NP [21-25]. COX dysregulation is also associated with aspirin intolerance, which is often accompanied by asthma and NP [22].
The pathogenesis of chronic rhinosinusitis with nasal polyposis is unknown. Chronic inflammation along with local tissue hypoxia may effect polyp's growth. Activation of Cycloxygenases may also be involved. COX-2 up-regulates in response to different stimuli including hypoxia. Its activation is associated with enhanced cell proliferation. Histologically, besides inflammatory cells, increased stromal fibrosis is seen in nasal polyposis. The aims of this study were to test whether hypoxia amplifies nasal polyp fibroblasts proliferation, whether treatment with various COX inhibitors could influence fibroblasts, and whether this effect may be modulated in response to different oxygenation conditions.
Polyp fibroblasts were incubated under hypoxic or normoxic conditions with or without NSAIDs at different concentrations for 12 or 24 hours. Cell proliferation was quantified using BrdU ELISA. Metabolic activity was evaluated using MTT assay. Cell death was measured using Annexin V staining and FACS scan.
No significant difference was found between proliferation of fibroblasts treated under hypoxia or normoxia. Cells incubated with indomethacin proliferated in a slightly enhanced manner compared with non-treated cells. Celecoxib inhibited fibroblast proliferation (P < .001) but did not influence cell survival. Metabolic activity of cells treated with celecoxib was significantly reduced (P < .003), unlike cells treated with indomethacin or rofecoxib.
Hypoxia does not affect fibroblasts proliferation. It may contribute to nasal polyposis pathogenesis in other ways. The anti-proliferative effect of celecoxib may be associated with cell cycle arrest rather than with pro-apoptotic activity. Celecoxib may be considered for treating nasal polyposis.
Reduced expression of COXs and production of prostaglandin E<inf>2</inf> in patients with nasal polyps with or without aspirin-intolerant asthma
2011, Journal of Allergy and Clinical Immunology
Citation Excerpt :
In one study COX-2 was reportedly upregulated in mast cells from airways of patients with AIA,23 whereas other studies reported no differences in expression of COX-2 in cells from upper25,26 or lower airways24 of patients with AIA, patients with ATA, or patients with chronic rhinosinusitis compared with healthy control subjects. The static characteristics of immunohistochemical analyses used in these studies23-26 might account for their discrepancies with findings from studies that measured COX-2 kinetics levels by means of immunoblotting or RT-PCR assays.21,35 Discrepancies might also result from the use of polyclonal versus mAbs to quantify levels of COX-2.36
Researchers have debated whether regulation of the COX enzymes (COX-1 and COX-2), which mediate production of prostaglandins (PGs), affects the pathogenesis of nasal polyps (NPs) and aspirin-intolerant asthma (AIA).
We investigated the roles of PGE₂, COX-1 and COX-2, and PGE₂ receptors in the development of NPs and AIA by measuring their expression in fibroblasts derived from nasal mucosa (NM) and NPs.
Fibroblasts were isolated from the NM of subjects without asthma who had septal deviation, turbinate hypertrophy, or both (control subjects, n = 7); NPs of aspirin-tolerant nonasthmatic patients (n = 7); and NPs of patients with asthma who were intolerant of aspirin (n = 7). Polyp samples were collected during endoscopic surgery. Cultures were stimulated with IL-1β (10 ng/mL) for 72 hours. We used ELISA, immunoblotting, and immunofluorescence analyses to measure secretion of PGE₂, expression of COX-1 and COX-2, and expression of the PGE₂ receptors EP1 to EP4.
Compared with NM from control subjects, PGE₂ concentrations were significantly lower in IL-1β–stimulated fibroblasts from patients with NPs who were tolerant to aspirin and even lower in polyps from patients with AIA. Similarly, IL-1β exposure induced the expression of COX-1 and COX-2 in fibroblasts from NM of control subjects, had only moderate effects on fibroblasts from NPs of aspirin-tolerant nonasthmatic patients, and almost no effect on fibroblasts from NPs of patients with AIA. IL-1β also induced expression of EP2 in fibroblasts from control NM but not in fibroblasts from NPs of aspirin-tolerant nonasthmatic patients or those with AIA.
Alterations in the COX pathway (ie, reduced production of PGE₂ and lack of upregulation of COX-1, COX-2, and EP2 under conditions of inflammation) are associated with NPs in patients with or without AIA.
Nasal Polyps: Pathogenesis and Treatment Implications
2011, Otolaryngologic Clinics of North America

View all citing articles on Scopus

^☆: Supported by grants from Fondo de Investigaciones de la Seguridad Social (95-0595 and 99-0133), Sociedad Española de Neumología y Cirugía Torácica (SEPAR), Sociedad Española de Alergología e Inmunología Clínica (SEAIC), and Generalitat de Catalunya (1998SGR112).

^☆☆: Reprint requests: Cesar Picado, Servei de Pneumologia I Allèrgia Respiratòria, Villarroel 170, 08036 Barcelona, Catalonia, Spain.

View full text

Mechanisms of AllergyCyclooxygenase 1 and cyclooxygenase 2 expression is abnormally regulated in human nasal polyps☆,☆☆

Abstract

Section snippets

Materials

Spontaneous regulation of Cox-1 and Cox-2 mRNA expression

Discussion

J Allergy Clin Immunol

Prostaglandins Leukot Essent Fatty Acids

Anal Biochem

J Biol Chem

Adv Immunol

Arch Biochem Biophys

Gastroenterology

FEBS Lett

J Biol Chem

Mol Cell Endocrinol

Cyclooxygenases: structural, cellular, and molecular biology

Annu Rev Biochem

Enhancement of acute allergic inflammation by indomethacin is reversed by prostaglandin E2: apparent correlation with in vivo modulation of mediator release

Proc Natl Acad Sci U S A

Inducible cyclooxygenase may have antiinflammatory properties

Nat Med

Impaired mucosal defense to acute colonic injury in mice lacking cyclooxygenase-1 or cyclooxygenase-2

J Clin Invest

Cyclooxygenase inhibition increases interleukin 5 and interleukin 13 production and airway responsiveness in allergic mice

Am J Respir Crit Care Med

Effect of inhibitors of prostaglandin synthesis and prostaglandins E2 and F2α on the immunologic release of mediators of inflammation from actively sensitized guinea-pig lung

J Pharmacol Exp Ther

Nasal polyps-a model of chronic respiratory mucosal inflammation

Clin Exp Allergy

Origin and structure of nasal polyps

Eicosanoids from biopsy of normal and polypous nasal mucosa

Rhinology

The pharmacological control of mediator release from human basophils and mast cells

Respiration

Mechanisms of Allergy
Cyclooxygenase 1 and cyclooxygenase 2 expression is abnormally regulated in human nasal polyps☆,☆☆

Enhancement of acute allergic inflammation by indomethacin is reversed by prostaglandin E₂: apparent correlation with in vivo modulation of mediator release

Effect of inhibitors of prostaglandin synthesis and prostaglandins E₂ and F_2α on the immunologic release of mediators of inflammation from actively sensitized guinea-pig lung