ISSN 0975-6299
Vol 3/Issue 1/Jan – Mar 2012
International Journal of Pharma and Bio Sciences
BIOCHEMISTRY
RESEARCH ARTICLE
ANTIOXIDANT ENZYMES, ANTIOXIDANT VITAMINS,GLUTATHIONE AND
URIC ACID LEVELS IN NASAL POLYP(S)
SHEETAL D1, SREEKANTHA*2, YOGESH B3 , AMARNATH M4 , MANJUNATH P5 ,KODLIWADMATH
M.V6, SREENIVAS RAO.K7, HARISH BHAT8AND VISHAL KALASKAR9.
1,5-
Department of ENT, AJIMS, Mangalore, Karnataka.
-Department of Biochemistry, NMC, Raichur, Karnataka.
3
-Department of pharmacology, Manipal University, KMC, Mangalore,Karnataka.
4
- Research Assistant, CCMB, Hyderabad, Andhra Pradesh.
2,6,7,8,9
SREEKANTHA
Department of Biochemistry, NMC, Raichur, Karnataka
ABSTRACT
Nasal polyp (NP) is an inflammatory pedunculated part of nasal mucosa. It is the most
common disease causing the nasal obstruction and sinusitis. Role of free radicals has been
proposed in the etiopathogenesis of many diseases including nasal polyp(s).Reactive oxygen
species (ROS) produce oxidative stress (OS) resulting in lipid and protein oxidation. The
amount of free radical injury or oxidative stress is assessed by altered levels of antioxidants.
So, we have estimated superoxide dismutase (SOD), catalase (CAT) as antioxidant
enzymes, ascorbic acid (Vit C) and tocopherol (Vit E) as antioxidant vitamins and glutathione
(GSH) and uric acid as antioxidants in 27 patients each in bronchial asthma nasal polyp
group (BANP group) and allergy nasal polyp group (ANP group) with 30 normal healthy age
matched controls by spectrophotometric method and uricacid in chemistry autoanalyser P800. Significantly decreased levels (p<0.001) of SOD, CAT, Vit C, Vit E, GSH and
significantly increased levels (p<0.001) of uric acid were found in both BANP group and ANP
group when compared to controls. Levels of all antioxidant parameters were altered more in
BANP group compared to ANP group. The documented results may be due to their utilization
during scavenging of ROS in disease process. Adequate supplementation of related
antioxidants may limit and prevent the disease process in both the groups.
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KEY WORDS
Nasal polyp(s) and Antioxidants.
INTRODUCTION
Nasal polyp (NP) is prolapsed pedunculated part
of nasal mucosa. It is very important to note that
nasal polyp is not a neoplasm. The most
common cause of nasal polyp is inflammation.
Nasal polyp is not a disease but it is the end
product of ongoing inflammation. There are two
types of nasal polyps; 1) Antrochonal polyp- it
arises from maxillary antrum and grows
backwards in the nose towards posterior nasal
passage. It is common in children and young
adults.2) Etamoidal polyp-they are multiple,
bilateral, arising from ethamoidal sinuses and
tend to proceed forwards[Delete] towards
external nasal orifices .They may occur at any
age. Etiology of NP includes infection, allergy,
bronchial asthma, cystic fibrosis, aspirin allergy,
non allergic rhinitis, selective IgA deficiency,
common
variable
immunodeficiency
etc.
Symptoms of NP include difficulty in breathing,
parosmia, running nose, snoring, chronic sinus
infection and dull headache. Complications of
NP includes infection, bleeding, chronic fungal
sinusitis which may the cause fungal meningitis
and in chronic and rare cases it may predispose
to malignancy. NP is diagnosed by
history,
clinical examination, nasal endoscopy or CT
scan-PNS.
Treatment
of
NP
includes
antihistamins, inhaled or oral steroids medication
and endoscopic sinus surgery for large polyps
(1-3).
ROS are intermediary metabolites that are
normally produced in the course of oxygen
metabolism. OS defines the imbalance between
formation of ROS and antioxidative defence
mechanism (4). Free radicals (FR) and ROS
have been implicated in most of human diseases
including cancer, organ failure, inflammatory
diseases, atherosclerosis etc. FR are highly
reactive species and hence they alter the
antioxidant defence system
(5).Study of
antioxidant enzymes, antioxidant vitamins along
with GSH and uric acid in literature is very
scanty, so the present study was designed to
estimate and compare the the[Delete] levels of
SOD and CAT as antioxidant enzymes, Vit C
and Vit E as antioxidant vitamins with GSH and
uric acid in BANP group and ANP group.
MATERIALS AND METHODS
Study Group: The Study group included 27
each patients between the ages 30 to 60 years.
They were classified into two groups:
.Nasal polyp(s) in Bronchial asthma patients
(BANP group).
1. Nasal poly(s) in allergy patients (ANP
group).
Blood samples were collected from patients were
attending outpatient department (OPD) and
admitted (IP) in ENT, Dermatology and General
Medicine wards were diagnosed to have nasal
poly(s)
with
clinical
and
radiological
investigations in the following hospitals;
1. A.J hospital. Kuntikan. Mangalore
2. ENT clinic, Sparsha, Kankanadi
3. KMC hospital. AC. Balmatta. Mangalore
4. Padmavathi hospital. Surathkal
5. Uma KMC hospital. Kasargod
6. KMC hospital. Attavar. Mangalore
7. Father Muller’s hospital. Kankanadi
Exclusion Criteria (In test group): Allergic rhinitis,
sinusitis, gross deviated nasal septum (DNS),
fungal allergic sinusitis, any benign and
malignant tumors of nasal cavity.
Control Group: The control group included 30
normal healthy subjects between the age group
of 30-60 years. While choosing the subjects for
test and control group, care was taken to
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eliminate those with habits like smoking, tobacco
chewing, alcohol consumption and also with
chronic inflammatory diseases like tuberculosis,
leprosy, SLE, rheumatoid arthritis, diabetes
mellitus etc, all of which play a vital role in
contributing to oxidative stress and hence alter
the various antioxidant parameters.
About the investigations, both the groups were
well informed and written consent was taken in a
proforma printed in a language known to
subjects. They were also informed about these
investigations that we are doing to know the
ethiopathogenesis of nasal polyp as the levels of
antioxidants in blood in both test groups [As they
were admitted or attending OPD’s of
Dermatology and General Medicine]. These
biochemical assays were analyzed on the same
day of blood collection from all groups. 5ml of
venous blood was collected in EDTA container
from median cubital vein or basilic vein of study
subjects under strict aseptic precautions. RBC
suspension was used for SOD and CAT
estimation, serum was used for Vit C, Vit E and
uric acid and plasma was used for GSH
estimation.
SOD activity was measured using the method of
Beauchamp and Fridovich (6).
CAT activity was estimated by Brunnan et al (7).
Vit C was measured by 2, 4- Dinitrophenyl
hydrazine method (8).
Vit E was measured using the method Baker H
et al (9).
GSH level in plasma was estimated by the
method of Beutler et al (10).
Uric acid was estimated by reduction of
phosphotungstic acid to blue phosphotungtus
acid, which is read at 700nm (11).
Statistical Analysis:
Statistical analysis was done by using KruskalWallis test.
RESULTS
All antioxidant parameters such as SOD, CAT,
Vit C, Vit E, and GSH were found very high
significantly (p< 0.001) reduced in both bronchial
asthma nasal polyp group( BANP group) and
allergy nasal polyp group (ANP group) but more
significant in BANP group. Uric acid level is very
high significantly (p< 0.001) increased in both
groups but more significant in BANP group when
compared with normal controls.
Levels of antioxidants in A.N.P and B.A.N.P groups compared to normal controls
Cases
SOD(U/g Hb)
p-value
CAT(U/g Hb)
p-value
Vit C(mg/L)
p-value
Vit E(mg/L)
p-value
GSH (mg/ml)
p-value
Uric acid(mg/dl)
p-value
Normal (Mean + SD)
A.N.P.G (Mean + SD)
B.A.N.P.G (Mean + SD)
30
7014.1413 + 1010.1231
5218.1600 + 3102.70
1.7690 + 0.2112
7.1280 + 2.8190
0.399 + 0.060
4.7994 + 1.0018
-
27
5148.03920 + 0249.728
<0.001
4161.14 + 652.19
<0.001
1.1010 + 0.1114
<0.001
3.010 + 1.192
<0.001
0.301 + 0.0610
<0.001
5.0798 + 0.2212
<0.001
27
4159.1202 + 0.592.211
<0.001
3812.0143 + 221.001
<0.001
0.9624 + 0.0119
<0.001
2.772 + 1.017
<0.001
0.274 + 0.0192
<0.001
8.1913 + 0.4256
<0.001
p< 0.001- very highly significant, SD- Standard Deviation
p-value: Probability of chance being the cause for the difference in the mean two group
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DISCUSSION
Nasal polyp (NP) is considered an inflammatory
condition in nasal and paranasal sinus cavities
and is commonly presented in ENT department.
The etiopathogenesis of nasal polyps is poorly
understood but the role of free radicals and
antioxidant defence is proposed as etiology of
the disease process. Inflammation, free radical
damage and oxidative stress have become
major health issues in past few decades. A byproduct of oxygen uptake by the body is the
formation of chemicals known as free radicals.
Free radicals have unpaired electrons. They
move around the body trying to get electrons
from other molecules. In the process of taking
the electrons, they damage the cells in a process
known as oxidation, causing oxidative stress.
Antioxidants are the natural substances they
neutralize the effects of radicals. The body
manufactures its own antioxidants. Some of
them are in the form of enzymes, such as
superoxide dismutase, catalase, glutathione
peroxidase etc, vitamins such as Vit C, Vit E,
Carotenes etc and other substances like
glutathione, uric acid, albumin, bilirubin etc.
Glutathione is an antioxidant that protects it from
free radical damage and carries out many other
essential functions, including detoxification,
repair of DNA, regulation of enzymes and
strengthening of our immune system. We have
chosen two of these enzymes, vitamins and
other two antioxidants for estimation.
In our study we reported decreased levels of
antioxidant enzymes (SOD & CAT), antioxidant
vitamins (Vit C & Vit E), GSH and increased
levels of uricacid. Taysi S et al (12)
demonstrated the decreased levels of various
antioxidant enzymes in the nasal polyp tissue. In
our study we have estimated all antioxidant
parameters in blood showing the altered levels in
accordance with the test results of the Taysi et al
study.
The current study demonstrates that there is
strong evidence related to oxidative stress
caused by free radicals in the etiopathogenesis
of nasal polyposis and antioxidants have a
definite role in free-radical-mediated tissue
damage in nasal polyposis and hence their
levels are decreased in serum to combat the
effect of free radical damage. Daqli M et al also
demonstrated that decrease levels of antioxidant
vitamins such as Vit C and Vit E and increased
oxidative stress in serum of nasal polyp patients
(13). Overproduction of free radicals results in
oxidative stress, a deleterious process that can
be an important mediator of damage to cell
structures, including lipids and membranes,
proteins, and DNA and hence the role of
antioxidants have a very notable role in oxidative
stress to neutralize the effects of free radicals;
therefore their levels are reduced in respective
disease condition where free radical etiology
persists (14). So in our observation in study, we
got altered antioxidant status indicating the
utilization of antioxidants during the free radical
defence. Uric acid levels is an end product of
purine metabolism and its levels were increased
in inflammatory diseases as purine catabolism is
increased in acute and chronic inflammation;
particularly in chronic inflammation and uric acid
levels were increased to combat the free radical
effects (15).Several studies on relationship of
oxidative stress, free radicals and antioxidants in
nasal polyp patients, concluded that decreased
levels of antioxidants indicated increased
oxidative stress causing the alteration of
antioxidants utilized for scavenging the free
radicals(16-18). Antioxidant parameters were
altered more in BANP and ANP group when
compared to controls but alteration is more in
BANP group because of oxidative stress is more
in bronchial asthma compared to allergy(1921).Several studies have demonstrated that the
supplementation of related antioxidants and
micronutrients in inflammatory process such as
bronchial asthma, allergy; they have noted that
the inflammatory process is very much reduced
and resulting in decreased disease symptoms
and complications (22-26)
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Hence, the supplementation of various related
antioxidants or supplementation of green leafy
vegetables, fruits, containing antioxidants such
as Vit C, Vit E, beta-carotene and trace elements
like zinc, copper etc which are required for the
functioning of antioxidant enzymes may limit the
disease process development or helps in aiding
the diagnosis, treatment and preventing the
complication and recurrence of disease process.
ACKNOWLEDGEMENT
Authors are extremely thankful for the kind
cooperation of Dr.Amareshwar, Dr.Rangaswamy
and Mr.Ramesh K during the study.
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