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Serum Glutathione as Biomarker for Oxidative

Stress in Health and Diseases

Article · March 2008

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 8002 ‫ المؤتمر العلمي العاشر‬/ ‫) عدد خاص‬11( ‫)المجلد‬1(‫ العدد‬/ ‫ العلوم‬/ ‫مجلة جامعة بابل‬

Serum Glutathione as Biomarker for Oxidative Stress in

Health and Diseases
Abdulsamie H. Alta'ee Mufeed J. Ewadh Zainab Mohammed Jasim
College of Medicine, Babylon University, Hilla P.O. Box 473, IRAQ.

Abstract
Background: Glutathione (GSH) an intracellular tripeptide has a central role in the protection against
oxidative stress and detoxification of xenobiotics.
Objective: To measure the involvement of GSH as antioxidant against oxidative stress due to several
type of diseases.
Patients and Method: GSH concentrations in the sera of twenty one patients (13 male, 8 female) with
Hodgkin's disease (HD), twenty three patients (13 male, 10 female) with non-Hodgkin's lymphoma
(NHL), thirty eight patients with breast cancer (BC), nine patients with bladder cancer, forty patients
(20 male, 20 female) with acute myocardial infarction (AMI), twenty nine patients (15 male,14 female)
with insulin dependent diabetes mellitus (IDDM) and fifty two (27 male, 25 female) healthy
individuals as a control were measured using Ellman's reagent.
Results: Compared with control, serum GSH concentrations found to be significantly (P<0.05)
decreased in both gender of patients with different type of diseases. Patients with IDDM, NHL, BC and
male with HD have a GSH concentration more than other types of diseases included in this study, and
the values of GSH concentration found to be higher in males than females .
Conclusion: Patients with IDDM, NHL, BC may exposure to oxidative stress, less than
patients with other type of diseases included in the present study.
‫كلوتاثايون المصل كدليل حيوي لإلجهاد التاكسدي في الصحة واألمراض‬
‫زينب محمد جاسم‬ ‫مفيد جليل عوض‬ ‫عبد السميع حسن الطائي‬
‫الخالصة‬
.‫الكلوتاثايون ببتيد ثالثي خلوي يلعب دو ار مركزيا في الحماية ضد االجهاد التاكسدي والتسمم بالمواد الغريبة عن الجسم‬
.‫يهدف البحث الى قياس تدخل الكلوتاثايون كمضاد لالكسدة ضد االجهاد التاكسدي والمتسبب من عدة انواع من االمراض‬
‫ اناث) مصابين‬8 ،‫ذكر‬31( ‫تم قياس تركيز الكلوتاثايون باستعمال كاشف المان في امصال دم واحد وعشرون مريضا‬
‫) وثمانية وثالثون مريضة‬NHL( ‫ اناث) مصابين بالورم اللمفاوي الالهوجكني‬31 ،‫ذكر‬31( ‫بمرض هوجكين وثالثة وعشرون مريضا‬
‫ انثى) مصابين باحتشاء العضلة القلبية‬01 ،‫ذكر‬01( ‫مصابة بسرطان الثدي وتسعة مرضى مصابين بسرطان المثانة واربعون مريضا‬
‫ ) اضافة الى اثنان وخمسون‬IDDM( ‫ اناث) بداء السكري المعتمد على االنسولين‬31 ،‫ذكر‬31( ‫الحاد وتسعة وعشرون مريضا‬
.‫ اناث) فردا صحيحا كمجموعة سيطرة‬01 ،‫ذكر‬02(
‫) في كال الجنسين‬p<0.05( ‫اظهرت النتائج عند مقارنتها بمجموعة السيطرة بان تركيز الكلوتاثايون انخفض بشكل معنوي‬
‫ مع مالحظة ان المرضى المصابين بداء السكري المعتمد على االنسولين و‬.‫عند المرضى باالنواع المختلفة من االمراض قيد الدراسة‬
‫بالورم اللمفاوي الالهوجكني وسرطان الثدي والذكور المصابين بمرض هوجكن بانهم يملكون تركيز كلوتاثايون اعلى من المرضى‬
.‫ ووجد تركيز الكلوتاثايون ليكون اعلى في الذكور من االناث‬،‫باالنواع االخرى من االمراض التي شملتها الدراسة‬
‫تستنتج الدراسة بان المرضى المصابين بداء السكري المعتمد على االنسولين و بالورم اللمفاوي الالهوجكني وسرطان الثدي‬
.‫من المحتمل ان يكونوا قد تعرضوا الى اجهاد تاكسدي اقل من االنواع االخرى من االمراض التي تضمنتها الدراسة الحالية‬

Introduction
Reactive oxygen species (ROS) are generated continuously during oxidative
metabolism. In order to avoid damage caused by ROS, such as DNA strand breaks,
lipid peroxidation and protein modification, mechanisms have been developed during
evolution which dispose of, or prevent the generation, of ROS. Increased production
of ROS and/or a decrease in the antioxidative capacity of cells causes oxidative stress
which can compromise essential cellular functions. (Dringen et al. 2000)

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 8002 ‫ المؤتمر العلمي العاشر‬/ ‫) عدد خاص‬11( ‫)المجلد‬1(‫ العدد‬/ ‫ العلوم‬/ ‫مجلة جامعة بابل‬

In the past few years, there has been increasing interest in the measurement of
thiols, glutathione (GSH) in particular, as indicators of oxidative stress.( Serru et al.
2001) Given the role of GSH in the protection against oxidative stress and
detoxification of xenobiotics, its availability in the reduced form (GSH) may be a key
factor in the maintenance of health. It has been established in several different animal
models, as well as in humans, that a decrease in GSH concentrations may be
associated with aging and the pathogenesis of many diseases, including rheumatoid
arthritis, muscular dystrophy, amyotrophic lateral sclerosis, AIDS, Alzheimer disease,
alcoholic liver disease, cataractogenesis, respiratory distress syndrome, progeria, and
Werner syndrome. (Rossi et al.2002 and James et al.2005)
Because blood GSH concentrations may reflect GSH status in other less
accessible tissues, measurement of both GSH and glutathione disulfide (GSSG) in
blood has been considered essential as an index of whole-body glutathione status and
a useful indicator of disease risk in humans. The reduced glutathione/ oxidized
glutathione ratio (GSH/GSSG) is used to evaluate oxidative stress status in biological
systems, and alterations of this ratio have been demonstrated in aging, cancer, HIV
replication, and cardiovascular diseases. ( Serru et al. 2001 and Rossi et al.2002)
GSH is also the main detoxifying agent in the body. It converts damaging
chemical substances (toxins) into harmless products that the body eliminates. Such
chemicals include cancer-producing substances, heavy metals, herbicides, pesticides,
smoke and other pollutants. Thus, GSH provides important protection against many
environmental hazards. The liver is particularly rich in glutathione for this purpose.
(Lands et al. 1999 and Douglas et al. 2002)
GSH is required in many intricate steps needed to carry out an immune
response. For example, it is needed for the lymphocytes to multiply in order to
develop a strong immune response, and for killer lymphocytes to be able to kill
undesirable cells such as cancer cells or virally infected cells. (Atalay et al.2002 and
Moellering et al. 2002)
In this study, we try to measure the involvement of GSH as antioxidant against
oxidative stress due to different type of diseases, and which of them consider to be the
most affective in the oxidative stress status.
This comparison has, to the best of our knowledge, never been tested in the
Iraqi population.

Patients and Methods

Patients
Patients subject to present study were grouped in to four main groups.
1- Forty patients with acute myocardial infarction (AMI), (20 male, 20 female).
2- Twenty nine patients with insulin dependent diabetes mellitus (IDDM), (15
male, 14 female).
3- Ninety one Patients with cancer :
a- Twenty one patients with Hodgkin's disease (HD), (13 male, 8 female).
b- Twenty three patients with non-Hodgkin's lymphoma (NHL), (13 male, 10
female).
c- Thirty eight female patients with breast cancer (BC).
d- Nine male patients with bladder cancer.
4- Control group which consist of fifty two (27 male, 25 female) healthy
individuals.

555
 8002 ‫ المؤتمر العلمي العاشر‬/ ‫) عدد خاص‬11( ‫)المجلد‬1(‫ العدد‬/ ‫ العلوم‬/ ‫مجلة جامعة بابل‬

Methods
Serum GSH was determined by using a modified procedure utilizing Ellman’s
reagent. 5,5´-Dithiobis (2-nitrobenzoic acid) (DTNB) is a disulfide chromogen that is
readily reduced by sulfhydryl group of GSH to an intensely yellow compound. The
absorbance of the reduced chromogen is measured at 412nm and is directly
proportional to the GSH concentration. (Burtis et al.1999)
Statistical Analysis
All values were expressed as mean ± standard deviation (SD) . Student's t-test
was used to estimate differences between the groups and differences were considered
significant when the probability was (p < 0.05).
Results
Compared with control ( 26.20 ± 7.52 and 19.22±4.94) for males and females
respectively, serum GSH concentrations found to be significantly (P<0.05) decreased
in both gender of patients with different type of diseases, as shown in Figure 1.
Patients with IDDM, NHL, BC and male with HD have a GSH concentration
more than other types of diseases included in this study.
Levels of GSH concentration found to be higher in males than females.
40

35

30
26.20
25
GSH (micro M)

19.22
20
16.17 16.65
15.62 15.58 15.68
14.82
15 12.83
12.02 12.19 11.88

10

0
AMI M AMI F DM 2 M DM 2 F NHL M NHL F HD M HD F bladder M breast F healthy M healthy F

M = male
F = female
Figure 1 GSH concentration (µM) in Patients with different type of disease and
healthy individuals. M=male, F=female
Discussion
There is significant evidence that the pathogenesis of several
neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease,
Friedreich's ataxia, amyotrophic lateral sclerosis, diabetes mellitus, alcoholic liver
disease, AIDS, acute hemorrhagic gastric crosions and cataracts, may involve the
generation of ROS and mitochondrial dysfunction. (Jörg et al. 2000 and Richie et al.
1996)
GSH, a ubiquitous cellular antioxidant, play a crucial role in defense against a
variety of disease and both exogenous and endogenous insults. Its functions include
the detoxification of xenobiotics, carcinogens, free radicals, and peroxides; regulation
of immune function; and maintenance of protein structure, function, and turnover.
(Richie et al. 1996)

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 8002 ‫ المؤتمر العلمي العاشر‬/ ‫) عدد خاص‬11( ‫)المجلد‬1(‫ العدد‬/ ‫ العلوم‬/ ‫مجلة جامعة بابل‬

Considerable evidence demonstrates that redox signaling mechanisms function

in cell regulation and growth control. GSH is the major low molecular weight thiol in
cells and plays a central role in controlling cellular thiol/disulfide redox state, which is
essential for normal redox signaling. (Yvonne et al.2002) As shown in Figure 2.

Figure 2 Function of GSH as an antioxidant. GSH reacts nonenzymatically with

radicals (R·) and is the electron donor for the reduction of peroxides (ROOH) in the
reaction catalyzed by GSH peroxidase (GPx.) GSH is regenerated from GSSG by
GSH reductase (GR) which uses NADPH as cofactor. {Figure taken from (Dringen et
al. 2000)}

Numerous laboratory studies have suggested that Blood and serum GSH
concentrations may serve as an indicator of GSH status and, thus disease risk in
human subjects. (Dringen et al. 2000 and Morrison et al.1999) In this respect, we try
to investigate the involvement of GSH as antioxidant against oxidative stress due to
diseases.
The results of present study are agreed with results of previous studies in
which significant depletion in GSH levels had been found. (Dringen et al. 2000 and
Yvonne et al.2002)
Compared with healthy controls, GSH concentrations were found to
significantly decrease in sera of patients with different types of cancers, IDDM and
AMI in the present study.
In this study, GSH concentration found to be higher in males than females in
patients and control groups. Another studies has reported that GSH concentrations
were 8 –10 % greater in smokers than in nonsmokers, and no sex differences. (Richie
et al. 1996) These results disagree with present study.
Patients with IDDM, NHL, BC and male with HD have a GSH concentration
more than other types of diseases included in this study.
Oxidative stress may be important in the development of coronary artery
disease because the heart, in contrast to the liver or lung, is relatively poorly defended
against oxidative stress. The levels of oxidized GSH in serum serve as an index of
myocardial oxidative stress during and after reperfusion. (Morrison et al.1999) This
may explain why patients with AMI have low levels of reduced GSH.
The depletion of GSH levels in the present study, when compared with healthy
control supports the hypothesis that considers GSH a protective factor against the
development of deferent types of diseases.
In conclusion the patients with IDDM, NHL, BC may exposure to oxidative
stress, less than patients with other type of diseases included in the present study.

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 8002 ‫ المؤتمر العلمي العاشر‬/ ‫) عدد خاص‬11( ‫)المجلد‬1(‫ العدد‬/ ‫ العلوم‬/ ‫مجلة جامعة بابل‬

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