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Mechanisms of Male Infertility: Role of

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Current Drug Metabolism, 2005, 6, 000-000

Mechanisms of Male Infertility: Role of Antioxidants

Salah A. Sheweita1,*, Abdulkarim M. Tilmisany2 and Hussein Al-Sawaf1
1

Department of Clinical Biochemistry, 2 Department of Pharmacology, Faculty of Medicine, Taibah University, Saudi
Arabia
Abstract: Defective sperm function is the most common cause of infertility, and until recently, was difficult to evaluate
and treat. Mammalian spermatozoa membranes are rich in poly unsaturated fatty acids and are sensitive to oxygen induced
damage mediated by lipid peroxidation. Hence, free radicals and reactive oxygen species [ROS] are associated with oxidative stress and are likely to play a number of significant and diverse roles in reproduction. The excessive generation of
reactive oxygen species by abnormal spermatozoa and by contaminating leukocytes [leukocytospermia] has been identified as one of the few defined etiologies for male infertility. Moreover, and environmental factors, such as pesticides, exogenous estrogens, and heavy metals may negatively impact spermatogenesis since male sperm counts were declined. In
addition, aging is also likely to further induce oxidative stress. Limited endogenous mechanisms exist to reverse these
damages. In a normal situation, the seminal plasma contains antioxidant mechanisms which are likely to quench these
ROS and protect against any likely damage to spermatozoa. However, during genitourinary infection/inflammation these
antioxidant mechanisms may downplay and create a situation called oxidative stress. Assessment of such oxidative stress
status [OSS] may help in the medical treatment of male infertility by suitable antioxidants. The cellular damage in the semen is a result of an improper balance between ROS generation and scavenging activities. Therefore, numerous antioxidants such as vitamin C, vitamin E, glutathione, and coenzyme Q10, have proven beneficial effects in treating male infertility. A multi-faceted therapeutic approach to improve male fertility involves identifying harmful environmental and occupational risk factors, while correcting underlying nutritional imbalances to encourage optimal sperm production and
function.

Key Words: Infertility, oxidative stress enzymes, glutathione, selenium, free radicals.
1. INTRODUCTION
Infertility has been a major medical and social preoccupation since the dawn of humanity. Defective sperm function
is the most prevalent cause of male infertility and a difficult
condition to treat [1]. Many environmental, physiological,
and genetic factors have been implicated in the conditions of
poor sperm function and infertility. It is important to identify
the factors which affect normal sperm function. However,
many men who demonstrate normal parameters on standard
semen analysis remain infertile [2]. This suggests that the
routine semen analysis such as measurement of seminal volume, spermatozoal motility, density, viability and morphology does not necessarily provide complete diagnostic information [3]. The assumption that free radicals can influence
male fertility has received substantial scientific support [4].
The proposed mechanism for loss of sperm function upon
oxidative stress has been shown to involve excessive generation of reactive oxygen species [ROS] [5].
1.1 Factors Affecting Male Fertility
1.1.1. Oxidative Stress and Reactive Oxygen Species
Most studies during the last decade have implicated oxidative stress as a mediator of sperm dysfunction. In the
*Address correspondence to this author at the Department of Clinical Biochemistry, Faculty of Medicine, PO Box 30001, Taibah University,
Madinah, Saudi Arabia; Tel: [04]-8158598; Fax: [04]-8461407;
E-mail: sheweita@hotmail.com

1389-2002/05 $50.00+.00

etiology of male infertility, there is growing evidence that

damage inflicted to spermatozoa by reactive oxygen species
[ROS] plays a key role [5, 6]. The spermatozoa have a high
content of polyunsatured fatty acids within the plasma membrane and a low concentration of scavenging enzymes within
the cytoplasm and they are susceptible to the peroxidation in
the presence of elevated seminal reactive oxygen species
levels [7-9]. To counteract the effects of reactive oxygen
species, spermatozoa and seminal plasma possess a number
of antioxidant systems. Spermatozoa possess a low amount
of cellular ROS defense system that consists of catalase, superoxide dismutase, glutathione peroxidase, and vitamin E.
In contrast, seminal plasma is well endowed with antioxidant
buffer capacity [10, 11]. The protective system includes
chain-breaking antioxidants capable of reducing oxidant
radical levels and blunting the propagation of free radical
chain reactions. Some studies have shown that infertile men
have an impaired seminal plasma nonenzymatic antioxidant
capacity than fertile ones, suggesting an association between
decreased total antioxidant capacity and male infertility [12,
13].
Previous studies have demonstrated that the spermatozoal
reactive oxygen species levels are higher in men with
varicocele, suggesting that sperm dysfunction in varicocele
[VAR] is in part related to oxidative stress [13, 14]. VAR
patients represent an interesting model since they exhibit an
enhanced ROS generation and high levels of nitric oxide,
which is related to ROS generation [15]. Moreover, infertile
men with varicoceles showed significantly increased sper 2005 Bentham Science Publishers Ltd.

Current Drug Metabolism, 2005, Vol. 6, No. 5

matozoal DNA damage that appears to be related to high

levels of oxidative stress [OS] in semen. Varicoceles are
found in approximately 15% of the general population and in
19%41% of men presenting for infertility investigations
[16-18]. The incidence of varicoceles in men with secondary
infertility is about 70%80% [19, 20]. These observations
suggested that varicoceles could cause a progressive decline
in fertility. Many patients with varicoceles also experience
altered spermatogenesis, which has been attributed to many
factors, including reflux of toxic metabolites from adrenal or
renal origin, disturbed hormone status, spermatic venous
hypertension, testicular hypoxia secondary to stasis, and abnormal temperature regulation [21]. Fujisawa et al. [1988]
found a significant reduction in levels of DNA polymerase in
extracts of testicular tissues from infertile men with
varicoceles [22]. As a result, they suggested that the decrease
in DNA polymerase activities might have deleterious effects
on spermatogenesis in patients with varicoceles. Another
factor that may lead to sperm DNA damage in these patients
is seminal oxidative stress [OS]. It has been found that high
levels of seminal OS and reduced total antioxidant capacity
[TAC] were detected in fertile and infertile men with a clinical diagnosis of varicoceles [23]. Increased levels of seminal
OS have been correlated with sperm dysfunction through
different mechanisms that include lipid peroxidation of
sperm plasma membrane and impairment of sperm metabolism, motility, and fertilizing capacity [24]. In addition, OS
has been shown to affect the integrity of the sperm chromatin
and to cause high frequencies of single and double DNA
strand breaks [25]. Previous studies have been indicated that
increased sperm nuclear DNA damage strongly and negatively affects natural fertility [26, 27]. Furthermore, it has
been reported that sperm chromatin/DNA is an independent
measure of sperm quality that may have better diagnostic
and prognostic capabilities than standard sperm parameters
[concentration, motility, and morphology] [28]. The cellular
damage in the semen is a result of an improper balance between ROS generation and scavenging activities. The scavenging potential in gonads and seminal fluid is normally
maintained by adequate levels of antioxidants superoxide
dismutase [SOD], catalase [CAT], glutathione peroxidase
[GPX], and glutathione reductase [GR]. This balance can be
referred to as oxidative stress status [OSS] and its assessment may play a critical role in monitoring sperm damage
and infertility. A situation in which there is a shift in this
ROS balance towards pro-oxidants, because of either excess
ROS or diminished anti-oxidants, can be classified as positive oxidative stress status [OSS]. However, assessment of
OSS, or a similar paradigm when monitored more objectively, would be a good indicator of sperm damage caused
by oxidative stress [29]. Chronic asymptomatic genitourinary inflammation can be regarded as a condition with positive OSS, which may be the real cause of idiopathic infertility in such patients. Superoxide dismutase [SOD] may directly act as antioxidant enzymes involved in the inhibition
of sperm lipid peroxidation [LPO] [30]. A high reduced glutathione [GSH]/ oxidized glutathione [GSSG] ratio will help
spermatozoa to combat oxidative insult [31]. It seems that
the role of these biological antioxidants and their associated
mechanisms is important in the treatment of infertility.

Sheweita et al.

1.1.2 Lipid Peroxidation of Spermatozoa

The presence of leukocytes in semen has been associated
with severe male infertility cases [7, 32, 33]. There has been
much speculation as to whether the origin of ROS in semen
is from spermatozoa or from infiltrating leukocytes [34, 35].
It has been reported that the leukocyte free percoll fractions
of semen samples obtained from nonazoospermic infertile
men generate detectable levels of ROS when compared to
the semen of normal and azoospermic men suggesting that
damaged spermatozoa are likely to be the source of ROS [7,
35]. Also, higher levels of ROS were correlated with a decreased number of motile sperm [7].
Mammalian spermatozoa are very susceptible to attack
by reactive oxygen species [ROS] attack since they are rich
in polyunsaturated fatty acids, which results in decreased
sperm motility, presumably by a rapid loss of intracellular
ATP leading to decreased sperm viability, and increased
morphology defects with deleterious effects on sperm capacitation and acrosome reaction. Lipid peroxidation of
sperm membrane is considered to be the key mechanism of
ROS-induced sperm damage leading to infertility [36, 37].
The most common types of LPO are: [a] nonenzymatic
membrane LPO, and [b] enzymatic [NADPH and ADP dependent] LPO. The enzymatic reaction involves NADPHcytochrome P-450 reductase [38]. Recently, it has been
found that cytochrome P450 4A was responsible for the generation of free radicals in experimental animals and human
tissues [39, 40]. In spermatozoa, production of malondialdehyde [MDA], an end product of LPO, has been reported
[41]. Formation of MDA can be assayed by the thiobarbituric acid [TBA] reaction which is a simple and useful diagnostic tool for the measurement of LPO for in vitro and in
vivo systems [42].
The human epididymis, the optimal site of sperm storage
and maturation, also has capacity to be protected from oxidative attack. Although the epididymis is able to synthesize
and secrete extracellular superoxide dismutase [SOD], concentrations of this and other antioxidants such as glutathione
peroxidase [GPX] and glucose -6-phosphate dehydrogenase
[GPD] are similar in semen from normal and vasectomized
men [43]. This indicates that the epididymal contribution of
antioxidants to semen may be slight, but does not preclude
important protective roles within the epididymis itself. The
existence of an epididymal-specific form of glutathione peroxidase and a high concentration of SOD within the epididymis would support this. Potts et al. [1999] [44] compared
seminal plasma from normal and vasectomized men for total
antioxidant activity, and thiol, ascorbate and uric acid content, individual antioxidants that are abundant in human semen [12]. Total antioxidant and thiol concentrations decreased in vasectomized men whereas ascorbic acid and uric
acid concentrations were unchanged and are therefore not
key exports of the epididymis. The increase in thiols suggests accumulation of thiol-containing compounds such as
GSH in the epididymal lumen. The lower total antioxidant
concentration reported in vasectomized men interestingly
mirrors that reported in infertile men. The epididymis therefore both contributes to seminal plasma and possesses region-specific antioxidant activity which may be important

Mechanisms of Male Infertility

for protection during epididymal storage and acquisition of

sperm functional competence. It has been found that removal
of the male accessory sex glands [ASG], which secrete a
variety of antioxidants, results in increased oxidative damage
in the form of DNA fragmentation, most notably in the less
mature sperm in the caput epididymis of golden hamsters
[45]. ROS can also induce oxidation of critical -SH groups in
proteins and DNA, which will alter structure and function of
spermatozoa with an increased susceptibility to attack by
macrophages [46]. As ASG removal also results in delayed
pronuclear formation at fertilization and increased embryonic wastage, this indicates the importance of ASG secretions in protecting sperm during the epididymal maturation
process. Thus, epididymal antioxidant status and degree of
oxidative stress may be important determinants of male fertility. In addition, the redox status of human spermatozoa is
likely to affect phosphorylation and ATP generation with a
profound influence on its fertilizing potential [47]. Aitken et
al. showed that stimulation of endogenous NADPHdependent ROS generation in human sperm appears to regulate acrosome reaction via tyrosine phosphorylation [48]. In
general, the oxidizing conditions increase tyrosine phosphorylation with enhanced sperm function while reducing
conditions have the opposite effect. These findings support
the concept that a critical level of ROS plays an important
role in spermatic vital functions. However, problems arise
only if excessive production of ROS and/or depletion of antioxidant system occur.
1.1.3 Nitric Oxide
Nitric oxide radical [NO.] and reactive nitrogen species
[RNS] have been found to have biological roles in mediating
many cytotoxic and pathological events [42]. Synthesis of
NO could contribute to poor sperm motility and function and
may lead to infertility [49]. RNS like ROS, may normally be
useful for maintaining sperm motility but can be toxic in
excess [50]. Other RNS such as nitrogen dioxide [NO.2]
radical and peroxynitrite [ONOO.] anion are considered to be
damaging agents. The primary mechanism of nitric oxide induced sperm damage is likely to be inhibition of mitochondrial respiration and DNA synthesis [51] or through its
interaction with superoxide anions and formation of peroxynitrite anion, which when protonated, decomposes to
form hydroxyl ion [OH.] and nitrogen dioxide [NO2.], both
of which are cytotoxic agents [52].
1.1.4 Aging and Oxidative Stress
The declining sperm count over the last two generations
is increasing in industrialized societies [53]. In addition,
some forms of infertility are caused by age related degenerative disorders of the testis. The decline in sperm numbers is
considered to be associated with male fetal and/or neonatal
exposure to increased level of the environmental estrogen.
The idiopathic male infertility may also be explained as a
form of premature or differential aging of the testis induced
by ischemia and oxidative stress associated with defective
mitochondrial genome that controls the oxidative phosphorylation [47]. Presence of retained cytoplasmic droplets
on spermatozoa due to imperfect spermiation in aging testis
may be a sign of' reduced fertility. It is known that lipid peroxidation [LPO] and midpiece anomalies are linked [54],
and that the increased rate of LPO and creatine kinase [CK]

Current Drug Metabolism, 2005, Vol. 6, No. 5

activity in immature sperm is due to incomplete cytoplasmic

extrusion during terminal spermatogenesis [55]. In addition,
Sertoli cells abnormality in infertile men may well be central
to the development of spermatogenic failure due to faulty
spermiation and may be related to genetic defects, oxidative
stress, or even aging of the gonads. Decreased vascularity,
increased spermatogenic failure, and reduced sperm output
occur with senescence in a variety of animal species and in
humans [56]. Degenerated germ cells are presumably phagocytosed by Sertoli cells, resulting in lipid accumulations that
increase progressively with age. Increased levels of lipofuscin and lipid are seen intracellularly, suggesting presence of
mitochondrial dysfunction possibly compounded by oxidative stress in the older population [57]. These degenerative
changes in gonads associated with accumulation of lipofuscin pigment and multiple nuclei are considered to be due to
ROS-induced lipid peroxidation with age [57]. Most of these
changes are strikingly similar to that seen in men with idiopathic testicular failure, probably due to induced gonadotoxicity.
1.1.5 Environmental Pollutants
There is a growing body of evidence supporting the idea
that sperm counts have declined considerably over the last
50 years. Carlsen et al. analyzed a total of 61 studies including 14, 947 men from the years 1930 to 1991, for mean
sperm density and mean seminal volume [58]. Their results
showed a significant decline in mean sperm density from
113 million/ml in 1940 to 66 million/ml in 1990 [p<0.0001].
Seminal volume decreased from an average of 3.40 ml to
2.75 ml [p=0.027] [59, 60]. This demonstrated a 20-percent
drop in volume and a substantial 58-percent decline in sperm
production in the last 50 years. Three other reports also
found semen quality has been declined among donors over
the last 20 years because the decline in sperm production is
due to a combination of environmental, lifestyle, and dietary
factors [59, 61]. It has been suggested that there are environmental reasons for deteriorating sperm quality, including
occupational exposure to various chemicals, heat, radiation,
and heavy metals [59, 62]. In addition, exposure to environmental estrogens and pesticides has been linked to alterations
in spermatogenesis. Another environmental concern with
infertility is the toxic effects of heavy metals on sperm quality and production. Heavy metals and chloro-organic compounds were found to influence female fertility at every
phase of reproduction. They may induce hormonal disorders,
preventing ovulations and pregnancies. There is the beginning of evidence that complications of pregnancy may be
related to pollution levels surrounding industrial plants. Reproductive health is affected through chromosome damage
and cell destruction, prenatal death, altered growth, fetal
abnormalities, postnatal death, functional learning deficits,
and premature aging. Men who are exposed to high levels of
lead may be at increased risk of becoming infertile as studies
have shown that higher lead levels interfere with the ability
of the sperm to bind to the egg and with its ability to fertilize
the egg [60]. Exposure to the common industrial chemical,
trichloroethylene [TCE], commonly found in adhesives, lubricants, paints, varnishes, paint strippers, pesticides, spot
removers and rug cleaning fluids, has been shown to adversely affect the normal development of sperm and cause
infertility [60]. Endocrine disrupting chemicals have been

Current Drug Metabolism, 2005, Vol. 6, No. 5

Sheweita et al.

hypothesized to influence male reproductive health [63]. The

suggested mechanism of action for these xenobiotics is their
capacity to interact with steroid hormones receptors, in particular those for estrogens and androgens [64].

smoke have a small but clinically significant increase in the

risk of spontaneous abortion [74-85].

Environmental factors such as chemical pollutants

blamed for declining sperm quantity and quality [65, 66].
The costs of environmental injury to reproduction include
subfertility, intrauterine growth retardation, spontaneous
abortion, and various birth defects. Developed country's primary threats are from chemical pollution, radiation, and
stress. Benzo[a]pyrene [B[a]P], a major environmental pollutant and component of cigarette smoke, is both carcinogenic and atherogenic in experimental models [67, 68].
Chronic administration of B[a]P did result in complete infertility in female birds, concomitant with grossly visible
changes in ovarian appearance. These results clearly show
that long-term dosing with B[a]P alters ovarian structure and
function in treated birds. Thus, B[a]P-induced atherogenicity
in female pigeons may be a consequence of an alteration in
estrogen production or antiestrogenic properties of B[a]P at
the level of the arterial wall and may serve as a highly useful
animal model to examine the well-known rapid development
of atherosclerosis in postmenopausal women [69]. In order
to assess the burden of environmental pollutants in the genital tract, 12 different chlorinated hydrocarbons were determined in 152 samples of follicular, seminal and cervical
fluids of patients. It has been showed that considerable concentrations of chlorinated hydrocarbons were present in parts
of the reproductive system, and that these compounds accumulate in the reception zone for spermatozoa [70]. Occupational exposure to polycyclic aromatic hydrocarbons was
significantly associated with higher levels of PAH-DNA
adducts and these results suggested the role for DNA damage in infertility [71]. Severe DNA damage, which might
prevent egg fertilization or the development of the embryo,
could be a cause of infertility. Therefore, [PAH]-DNA adducts were used as marker of sperm genotoxicity and infertility [71].

In general, antioxidants are compounds which dispose,

scavenge, and suppress the formation of ROS, or oppose
their actions. The major antioxidants are vitamin A, vitamin
E, beta-carotene, vitamin C and the trace mineral selenium.
A number of nutritional therapies have been shown to improve sperm counts, and sperm motility, including carnitine,
arginine, zinc, selenium, and vitamin B-12. Antioxidants like
vitamin C, vitamin E, glutathione, and coenzyme Q10, have
also proven beneficial in treating male infertility [86-90].
They are constantly rendering free radicals harmless. To
protect the cells and organ systems of the body against reactive oxygen species, humans have evolved a highly sophisticated and complex antioxidant protection system [91]. It
involves a variety of components, both endogenous and exogenous in origin, that function interactively and synergistically to neutralize free radicals [91].

Although cigarette smoking is a widely recognized health

hazard and a major cause of mortality, people continue to
consume cigarettes on a regular basis. Approximately onethird of the world's population smokes cigarettes daily [71,
73]. Needless to say, a number of nonsmokers are also negatively affected when they inhale side-stream smoke from
burning cigarettes. Given the large number of men worldwide who smoke, and the fact that cigarette smoke is a
known somatic cell mutagen and carcinogen, there is a major
concern that smoking may adversely affect male reproductive health [74, 75]. A number of studies have shown that
smoking has a detrimental effect on sperm quality, most significantly sperm concentration, motility, and morphology
[76-78]. In addition, cigarette smoking has been correlated
with poor sperm function in sperm penetration assays [Close
et al., 1990]. Furthermore, paternal smoking has been associated with a significant increase in the percentage of spermatozoa with DNA damage [44, 80, 81]. Epidemiological
studies in women of reproductive age have shown that cigarette smoking has a dose-related effect that can delay time to
conception by 2 months [72] and advance the start of menopause by 2 years [78]. It has been suggested that women who

1.2 Antioxidants and Infertility

Humans have evolved a highly sophisticated and complex antioxidant protection system to protect the cells and
organ systems of the body against reactive oxygen species
[92]. It involves a variety of components, both endogenous
and exogenous in origin, that function interactively and synergistically to neutralize free radicals. These components
include: (a) Nutrient-derived antioxidants like ascorbic acid
(vitamin C), tocopherols and tocotrienols (vitamin E), carotenoids, and other low molecular weight compounds such as
glutathione and lipoicacid [92], (b) antioxidant enzymes,
e.g., superoxide dismutase, glutathione peroxidase, and glutathione reductase, which catalyze free radical quenching
reactions, (c) metal binding proteins, such as ferritin, lactoferrin, albumin, and ceruloplasmin that sequester free iron
and copper ions that are capable of catalyzing oxidative reactions, and (d) numerous other antioxidant phytonutrients
present in a wide variety of plant foods [92].
Many plant-derived substances, collectively termed
phytonutrients, or phytochemicals, are becoming increasingly known for their antioxidant activity. Phenolic
compounds such as flavonoids are ubiquitous within the
plant kingdom: approximately 3, 000 flavonoid substances
have been described [97]. In plants, flavonoids serve as protectors against a wide variety of environmental stresses
while, in humans, flavonoids appear to function as biological response modifiers. The broad therapeutic effects of flavonoids can be largely attributed to their antioxidant properties [98].
A number of studies have evaluated the role of vitamins
and antioxidants in male infertility, and, most, but not all, of
these studies have suggested a beneficial role for antioxidants therapy in the treatment of male infertility. Fraga et al.,
1991, demonstrated that dietary vitamin C has a beneficial
effects on the integrity of sperm DNA in male smokers [99].
In a small, placebo-controlled trial, Dawson et al. 1992,
found that supplemental vitamin C improved sperm quality
[100]. A number of investigators have showed that vitamin E
improved sperm quality [101-103]. In a small placebocontrolled trial, Lenzi et al. 1993, reported that intramuscular

Mechanisms of Male Infertility

glutathione improved sperm quality significantly. Controlled

and uncontrolled trials have demonstrated a beneficial effect
of -carnitine on male fertility [104, 105]. Folic acid and zinc
supplements [in combination] have been shown to increase
sperm counts in a placebo-controlled trial [106]. In contrast
to the above positive studies, Rolf et al.1999, reported that
no beneficial effect on sperm quality with the use of combined therapy with vitamins C and E in a small, placebocontrolled trial [107].
Coenzyme Q10 [CoQ10] is a component of the mitochondrial respiratory chain and plays a crucial role in energy
metabolism [35, 38, 87]. Furthermore, it is an important
liposoluble chain-breaking antioxidant associated with
membranes and lipoproteins. It has long been known that
CoQ10 biosynthesis is markedly active in testis and high
levels of its reduced form QH2 [ubiquinol] are present in
semen [108-110], which suggests a protective role as a scavenger in this biological system. There is evidence that sperm
cells with reduced motility also have a significant reduction
in the phospholipid pool, as well as phosphatidylethanolamine and phosphatidylcholine [PC] content, probably related
to a reduction in the antioxidant capacity of spermatozoa and
seminal plasma [111]. It has been demonstrated that reduced
levels of CoQ10 in the seminal plasma and sperm cells of
infertile men with idiopathic and varicocele-were associated
with asthenospermia [112]. On this basis, CoQ10 is indicated
as one of the compounds contributing to the total antioxidant
buffer capacity of semen and its reduction leads to an impairment of the system due to oxidative stress [113].
Whether the exogenous administration of CoQ10 could lead
to any modification of its content in semen or to any benefit
on sperm cell function still remains an interesting open
problem dietary implementation.
A deficiency of dietary selenium leads to immotile, deformed sperm and infertility in rats, whereas supplementation of the diet with selenium compounds has been associated with both beneficial and deleterious effects on sperm
function, depending on the chemical form of selenium [114].
Serum triiodothyronine decreased and thyroid-stimulating
hormone increased in the high-selenium group, suggesting
that altered thyroid hormone metabolism may have affected
sperm motility. Although this decrease in sperm motility
does not necessarily predict decreased fertility, the increasing frequency of selenium supplementation in the healthy
population suggests the need for larger studies to more fully
assess this potential side effect [115].
A decrease in levels of reduced glutathione [GSH] during
sperm production has been shown to cause disruption in the
membrane integrity of spermatozoa as a consequence of increased oxidative stress [116]. Intracellular glutathione levels
of spermatozoa are shown to be decreased in certain populations of infertile men [117-119]. Glutathione is not only vital
to sperm antioxidant defenses, but also is essential to the
formation of phospholipid hydroperoxide glutathione peroxidase - an enzyme present in spermatids which becomes a
structural protein in the mid-piece of mature spermatozoa.
Deficiencies of either substance can lead to instability of the
mid-piece, resulting in defective motility [120, 121]. Scavengers, such as glutathione can be used to treat these cases as
they can restore the physiological constitution of poly-

Current Drug Metabolism, 2005, Vol. 6, No. 5

unsaturated fatty acids [PUFA] in the cell membrane [122,

123]. In a double-blind cross-over study of 20 infertile men,
treatment with glutathione led to a statistically significant
improvement of the sperm quality [96, 124, 125].
Levels of reduced glutathione were significantly decreased in oligospermic and azoospermic group, and the reduction in azoospermic group [76.73%] was more pronounced than oligozoospermic group [62.07%]. The decrease in reduced glutathione levels in azoospermic and oligozoospermic conditions may cause disruption in the membrane integrity of spermatozoa as a consequence of increased
oxidative stress [35, 126].
Many studies have been reported on the role of super
oxide dismutase [SOD] as an antioxidant in reproductive
biology. SOD protects spermatozoa against spontaneous O2
toxicity and LPO [126, 127]. SOD and catalase also remove
[O2] generated by NADPH-oxidase in neutrophils and may
play an important role in decreasing LPO and protecting
spermatozoa during genitourinary inflammation [127]. Glutathione reductase then regenerates reduced GSH from oxidized form GSSG. A selenium-associated polypeptide, presumably glutathione peroxidase, has been demonstrated in
rat sperm mitochondria to play a significant role in this peroxyl scavenging mechanism and in maintaining sperm motility [128]. In addition, GSH peroxidase and GSH-reductase
may directly act as antioxidant enzymes involved in the inhibition of sperm LPO [125]. In this context, the gamaglutamyl transpeptidase [gama-GT], considered being present in the midpiece and acrosomal regions of spermatozoa
of certain mammalian species may further affect GSH content of oocyte at the time of sperm penetration.
1.3 CONCLUSIONS
Increased oxidative damage to sperm membranes [indicated by increased LPO], proteins, and DNA is associated
with alterations in signal transduction mechanisms that affect
fertility. Spermatozoa and oocytes possess an inherent but
limited capacity to generate ROS which may help the fertilization process. A variety of defense mechanisms encompassing antioxidant enzymes [SOD, catalase, glutathione
peroxidase and reductase], vitamins [E, C, and carotenoids],
and biomolecules [glutathione and ubiquinol] are involved in
biological systems. A balance between the benefits and risks
from ROS and antioxidants appears to be necessary for the
survival and normal functioning of spermatozoa. An assay
system for the evaluation of oxidative stress status [OSS]
may aid the clinician in the assessment of fertility status of
both male and female partners.
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