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Miwa et al., J Otol Rhinol 2015, S1:1
http://dx.doi.org/10.4172/2324-8785.S1-004
Journal of Otology &
Rhinology
Research Article
A SCITECHNOL JOURNAL
Peroxide Tone in Human Inferior
Nasal Turbinate with Allergy
Peroxide; TBA: Thiobarbituric Acid; TBA-RS: Thiobarbituric Acid
Reactive Substances; RAST: Radioallergosorbent Test; TBA:
Thiobarbituric Acid
Masato Miwa1*, Noritsugu Ono1, Daisuke Sasaki1,
Shiozawa1, Mayumi Miwa2 and Katsuhisa Ikeda1
Introduction
1Department
Akihito
of Otorhinolaryngology, Juntendo University Faculty of Medicine,
Tokyo, Japan
2Harimazaka
Clinic, 4-20-2 Koishikawa, Tokyo, Japan
*Corresponding
113-8421,
author: Masato Miwa, MD, 2-1-1 Hongo, Bunkyo-ku, Tokyo
Japan, Tel: +81-3-5802-1094; Fax: +81-3-5689-0547; E-mail:
[email protected]
Rec date: Nov 17, 2014 Acc date: Jan 16, 2015 Pub date: March 07, 2015
Abstract
Background: The nose is chronically exposed to oxidative
stress, which can easily lead to reactive oxygen species
(ROS)-mediated damage and lipid oxidative damage of the
upper airway. ROS may also participate in various diseases,
including those of the airway, although many details are not yet
known.
ROS is generated by various enzymatic reactions and chemical
processes. Superoxide dismutase (SOD), catalase, and
glutathione
peroxidase
(GSH-Px)
are
representative
scavengers. ROS are also generated through arachidonic acid
cascades. One of the primary prostaglandins (PG), PGD2, is
the major PG in most types of tissue, including the nose. We
attempted to evaluate the peroxide tone by measuring these
factors.
Methods: A total of 42 Japanese patients with and without
nasal allergies were enrolled in this study. We determined the
contents of lipid peroxide (LPO) and PGD2, as well as the
activities of SOD, catalase, and GSH-Px of the anterior portion
of the mucosa of the inferior turbinate, obtained by inferior
turbinotomy.
The nose is chronically exposed to oxidative stress, leading to
reactive oxygen species (ROS)-mediated damage and lipid oxidative
breakage of the upper airway. Many researchers have demonstrated
that ROS can have a variety of physiological and deleterious effects
within the airway [1]. It appears that ROS may participate in various
diseases including those of the nose, although many details are not yet
known.
ROS is a general term that includes a large variety of free oxygen
radicals, generated by various enzymatic reactions and chemical
processes. Superoxide dismutase (SOD, EC1.15.1.1) converts
superoxide anion to hydrogen peroxide. Catalase (EC1.11.1.6)
converts hydrogen peroxide into oxygen and water. Glutathione
peroxidase (GSH-Px, EC1.11.1.9) inactivates hydrogen peroxide. ROS
are also generated through arachidonic acid cascades. Peroxinitrite can
activate both cyclooxygenase -1 and -2 [2]. Studies have demonstrated
that prostaglandin (PG) D2is the major PG in most types of tissue,
representing 50-96% of the total primary PGs [3]. We previously
demonstrated the Immunohistochemical localization of PGD
synthetase in nasal mucosa [4].
We couldn't find any studies in the literature that have conducted a
comprehensive analysis of the metabolic pathway of ROS in human
inferior nasal turbinate, the front line of the respiratory tract. In order
to evaluate the relationship between the arachidonic acid cascade and
oxygen radical formation,and demonstrate the peroxide tone in
patients with allergic rhinitis, we investigated the contents of lipid
peroxide (LPO) and PGD2, as well as the activities of representative
antioxidant enzymes, such as SOD, GSH-Px and catalase in mucosa of
human inferior turbinate obtained from patients with and without
nasal allergy.
Methods
Abbreviations:
The subjects in this study were 42 patients in Juntendo University
Faculty of Medicineand Harimazaka Clinic, who was all underwent
bilateral inferior turbinotomy for chronic rhinitis with and without
nasal allergy,due to nasal obstruction. The subjects were between 20 to
59 years of age, of either gender. The diagnosis of allergic rhinitis was
based on clinical symptoms, a nasal smear test for eosinophils and
Immuno CAP for house dust. A total of 22 patients were diagnosed
with allergic rhinitis. The anterior portion of the mucosa of the
inferior turbinate was obtained by inferior turbinotomy under general
or local anesthesia from patients with and without nasal allergy. None
of the subjects had abnormal blood examination data, including serum
lipids. None of the subjects were taking systemic antihistamine or
steroids prior to the surgery. On removal, specimens were put into
liquid nitrogen and preserved in a deep freezer at − 80°C until the
biochemical analysis was conducted. Because the study dealt with
removed tissue and did not affect the patients themselves, the
institutional review board granted exemption from the formal
approval process.
ROS: Reactive Oxygen Species; SOD: Superoxide Dismutase; GSHPx: Glutathione Peroxidase; PG: Primary Prostaglandins; LPO: Lipid
LPO was measured using the fluorescent method with
malondialdehyde as a standard, as described by Ohishi [5]. LPO
Results: LPO and PGD2 contents increased significantly in the
nasal allergy group. No statistically significant difference
appeared in the activities of SOD, catalase, and GSH-Px were
demonstrated in the nasal allergy group compared with the
subjects without nasal allergy.
Conclusion: The imbalance of the peroxide tone in the nasal
mucosa caused by stimulation of the cyclooxygenase pathway
of the arachidonic acid cascade and ROS formation was
demonstrated. Moreover, the increased number of ROS was
not well metabolized in the nasal mucosa with allergy.
Keywords: Allergic rhinitis; Reactive oxygen species; Prostaglandin
D2; Nasal mucosa; Superoxide dismutase; Catalase, Glutathione
peroxidase; Lipid peroxide
All articles published in Journal of Otology & Rhinology are the property of SciTechnol and is protected by copyright
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Citation:
Miwa M, Ono N, Sasaki D, Shiozawa A, Miwa M, et al. (2015) Peroxide Tone in Human Inferior Nasal Turbinate with Allergy. J Otol Rhinol S1:1.
doi:http://dx.doi.org/10.4172/2324-8785.S1-004
contents were expressed as thiobarbituric acid (TBA) reactive
substances. The assay of the total SOD activity in the nasal mucosa was
performed using nitrous acid, as described by Oyanagui [6].The assay
of the GSH-Px was performed employing Hochstein's method. The
assay of the catalase was performed employing Thomson's method [7].
The extraction of the PGs was performed as described by Powell
[8]. The concentrations of the PGs were measured by HPLC-RIA [3].
Protein measurement was made by the Lowry method [9].
Data were expressed as mean ± SEM. The paired t test was used to
ascertain whether there was a statistically significant difference in the
peroxide tone between rhinitis with and without nasal allergy. The
chosen level of significance was 0.05 in all tests.
Results
The results for the LPO content were expressed as thiobarbituric
acid reactive substances (TBA-RS). The TBA-RS value was 2.08 ±
0.28nmol/mg protein in subjects with allergic rhinitis and 1.20 ± 0.19
nmol/mg protein in the subjects with non-allergic rhinitis, showing a
statistically significant difference (Figure 1). The total SOD activity
was 27.8 ± 2.3 ng/mg protein in subjects with allergic rhinitis and 30.2
± 2.1 ng/mg protein in subjects with non-allergic rhinitis (Figure 2).
No statistically significant difference was seen between the total
SOD activities in the subjects with allergic rhinitis, compared with the
subjects with non-allergic rhinitis.
Figure 2: The results of Total SOD activity in human nasal mucosa
with and without allergic rhinitis. The total SOD activity was 27.8 ±
2.3 ng/mg protein in subjects with allergic rhinitis and 30.2 ± 2.1
ng/mg protein in subjects with non-allergic rhinitis. No statistically
significant difference was seen between the total SOD activity in the
subjects with allergic rhinitis, compared with the subjects with nonallergic rhinitis.
GSH-Px activity was 43.7 ± 4.14 U/mg protein in subjects with
allergic rhinitis and 45.8 ± 2.23 U/mg protein in those with nonallergic rhinitis (Figure 3). No statistically significant difference was
seen between the GSH-Pxactivity in the subjects with allergic rhinitis,
compared with the subjects with non-allergic rhinitis.
Figure 1: The results concerning thelipid peroxide content in
human nasal mucosa with and without allergic rhinitis. LPO are
expressed as thiobarbituric acid reactive substance values. The
TBA-RS value was 2.08 ± 0.28 nmol/mg protein in subjects with
allergic rhinitis and 1.20 ± 0.19 nmol/mg protein in the subjects
with non-allergic rhinitis showing a statistically significant
difference.
Volume S1 • Issue 1 • S1-004
Figure 3: The results of GSH-Px activity in human nasal mucosa
with and without allergic rhinitis. GSH-Px activity was 43.7 ± 4.14
U/mg protein in subjects with allergic rhinitis and 45.8 ± 2.23
U/mg protein in those with non-allergic rhinitis. No statistically
significant difference was seenbetween the GSH-Pxactivity in the
subjects with allergic rhinitis, compared with the subjects with nonallergic rhinitis.
The catalase activity was 16.4 ± 2.54 U/mg protein in the subjects
with allergic rhinitis and 12.0 ± 1.01 U/mg protein in the subjects with
non-allergic rhinitis (Figure 4). No statistically significant difference
was seen between the catalase activities in the subjects with allergic
rhinitis, compared with the subjects with non-allergic rhinitis. The
PGD2 content was 4.67 ± 0.58 ng/g tissues in the subjects with allergic
rhinitis and 2.54 ± 0.27 ng/g tissues in those with non-allergic rhinitis
(Figure 5). A significantly higher level of PGD2 was found in the nasal
• Page 2 of 5 •
Citation:
Miwa M, Ono N, Sasaki D, Shiozawa A, Miwa M, et al. (2015) Peroxide Tone in Human Inferior Nasal Turbinate with Allergy. J Otol Rhinol S1:1.
doi:http://dx.doi.org/10.4172/2324-8785.S1-004
mucosa of the subjects with allergic rhinitis, compared with the
subjects with non-allergic rhinitis.
Figure 4: The results concerning thecatalase activity in human nasal
mucosa with and without allergic rhinitis. Thecatalase activity was
16.4 ± 2.54 U/mg protein in the subjects with allergic rhinitis and
12.0 ± 1.01 U/mg protein in the subjects with non-allergic rhinitis.
No statistically significant difference was seen between the catalase
activity in the subjects with allergic rhinitis compared with the
subjects with non-allergic rhinitis.
Figure 5: The results concerning theprostaglandin D2 content in
human nasal mucosa with and without allergic rhinitis. The PGD2
content was 4.67 ± 0.58 ng/g tissues in the subjects with allergic
rhinitis and 2.54 ± 0.27 ng/g tissues in those with non-allergic
rhinitis. A significantly higher level of PGD2 was found in the nasal
mucosa of the subjects with allergic rhinitis compared with the
subjects with non-allergic rhinitis.
Discussion
It is considered that various chemical mediators, including ROS, are
interrelated in the pathogenesis of nasal allergies. Moreover, many
enzymes and interact with each other in the metabolism of eicosanoid
Volume S1 • Issue 1 • S1-004
formation. A redox-active protein, thioredoxin, plays a crucial role in
the metabolism of the antioxidant system, regulating the reduction/
oxidation balance by scavenging ROS, which are implicated in the
mechanism of asthma [10]. In order to evaluate the effect of ROS in
the onset and development of diseases, it may be considered that a
comprehensive analysis of the metabolic pathway in the formation of
ROS is necessary.
Scheme 1: Schematic drawing of the formation and possible
interaction between oxygen species inhuman nasal mucosa. An
increase in the lipid peroxide (LPO, shown as LCCO here) and PG
(prostaglandin) D2 contents, and a decrease in activities of
superoxide dismutase (SOD ), catalase, and glutathione peroxidase
(GSH-Px) were demonstrated in the inferior turbinate of patients
with allergic rhinitis.
Immune modulation targeting critical Th2 effector molecules has
shown promise in allergic disease, particularly strategies directed
against IgE, but also prostaglandin D2 (PGD2) and leukotriene D4
(LTD4). Targeting such molecules may provide better benefit-to-risk
profiles over cytokine-directed therapies as they appear to be more
specific to allergic driven processes [11]. Matsuoka et al. [12] have
shown that PGD2 functions as a mediator of allergic asthma. In
addition to being produced in the lung, PGD2 may also play an
important role in other allergic disorders, such as allergic rhinitis and
atopic dermatitis. The DP receptor may thus represent a new
therapeutic target for the treatment of such allergic reactions.
According to Hemler [13], the formation of PG and ROS is regulated
by positive feedback control. He demonstrated that the balance
between the formation and removal of cellular LPO sets a peroxide
tone that can regulate the rate of PG formation in cells. Therapies
directed against specific effector molecules, such as immunoglobulin E
and prostaglandin D2, hold promise in the immune modulation of
allergic diseases, as do therapies targeting the IL-4/IL-13 receptor and
augmenting the Th1/Th2 balance with Toll-like receptor agonists [14].
We demonstrated that the PGD2 and LPO contents in the nasal
mucosa are higher in subjects with perennial allergic rhinitis,
compared with those with non-allergic rhinitis. High levels of
malondialdehyde, one of the metabolites of free radical-mediated lipid
peroxidation, were observed in nasal polyps.
• Page 3 of 5 •
Citation:
Miwa M, Ono N, Sasaki D, Shiozawa A, Miwa M, et al. (2015) Peroxide Tone in Human Inferior Nasal Turbinate with Allergy. J Otol Rhinol S1:1.
doi:http://dx.doi.org/10.4172/2324-8785.S1-004
On the other hand, the activities of SOD, GSH-Px and catalase in
the nasal mucosa were similar in the tissues obtained from subjects
with allergic rhinitis and those with non-allergic rhinitis.
It is well known that antioxidative enzymes (AOEs), such as catalase
(CAT), glutathione peroxidase (GPX), glutathione reductase (GR),
superoxide dismutase (SOD) and glutathione S-transferase (GST), are
of great importance in the mucosal defense against reactive oxygen
species[1]. Previous studies regarding the measurement of SOD
activities in nasal polyps are still controversial [15]. Cannady et al. [16]
reported that the enzyme activity of SOD was lower in the nasal polyps
than in the normal inferior turbinate. In contrast, another study
showed that SOD activity was unchanged in the nasal polyps,
compared with the normal middle turbinate. The pathogenesis of
nasal allergy is quite different from that of chronic Rhinosinusitis with
polyp. To evaluate the activities of antioxidant enzymes in the inferior
turbinate where allergic reaction may occur in the beginning of allergic
rhinitis, we demonstrated that SOD activity in the inferior turbinate
was statistically the same with and without nasal allergy. Several
different iso forms of antioxidant enzymes have been identified in
humans [17]. We evaluated the total activities of those enzymes, so it
remains unclear which isozymes are dominant. These results suggested
that the cyclooxygenase pathway of the arachidonic cascade is
stimulated and ROS formation is accelerated in human nasal mucosa
with allergies. This isconsistent with areport describing that combined
antihistamine and cyclooxygenase-inhibiting drugs were effective for
hay fever treatment [18]. Moreover, it may be considered that the
increased number of ROS is not well metabolized in nasal mucosa with
allergies.
The LPO contents in the nasal mucosa with chronic rhinosinusitis
were not increased compared with nasal mucosa without
rhinosinusitis, as reported by Friedman [19]. We have demonstrated
adecrease of the total SOD activity ineosinophilic and noneosinophilic sinusitis [20]. In contrast, we demonstrated increased
LPO in allergic rhinitis. These phenomena seem to be related to the
pathogenesis of allergic rhinitis. Consequently, the control of PGD2
production and ROS may thus be quite useful in the treatment of
allergic diseases involving the nose. It is hoped that further studies of
the enzymes metabolizing ROS will supply new information regarding
the mechanisms of allergies in the upper airway.
We conclude that the alteration of the peroxide tone in the nasal
mucosa caused by stimulation of the cyclooxygenase pathway of the
arachidonic acid cascade and ROS formation may be intimately related
to the pathogenesis of allergic rhinitis.
Conclusion
The imbalance of the peroxide tone in the nasal mucosa caused by
stimulation of the cyclooxygenase pathway of the arachidonic acid
cascade and ROS formation was demonstrated. Moreover, the
increased number of ROS was not well metabolized in the nasal
mucosa with allergies.
Acknowledgment
Authors are grateful to Dr. Yoshihiro Urade of Department of
Molecular Behavioral Biology, Osaka Bioscience Institute, Osaka,
Japan, for checking the design of this study.
Volume S1 • Issue 1 • S1-004
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Citation:
Miwa M, Ono N, Sasaki D, Shiozawa A, Miwa M, et al. (2015) Peroxide Tone in Human Inferior Nasal Turbinate with Allergy. J Otol Rhinol S1:1.
doi:http://dx.doi.org/10.4172/2324-8785.S1-004
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