Experimental and Toxicologic Pathology 63 (2011) 221–227

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Experimental and Toxicologic Pathology

journal homepage: www.elsevier.de/etp

Influence of vitamin C supplementation on lead-induced histopathological

alterations in male rats
Mahmoud Shaban El-Neweshy a, Yasser Said El-Sayed b,n
a
Department of Veterinary Pathology and Parasitology, Faculty of Veterinary Medicine, Alexandria University, Egypt
b
Department of Veterinary Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, Alexandria University (Damanhur Branch), Behera Province, Egypt

a r t i c l e in fo abstract

Article history: This study is intended to evaluate the efficacy of vitamin C (VC) in ameliorating the detrimental effects
Received 30 October 2009 of long-term lead intoxication on the liver, kidneys, brain and testes as assessed by histopathology. A
Accepted 14 December 2009 total of forty male Wistar rats (six-weeks-old) was divided into 4 groups: control group; lead-acetate
(PbAc)-treated group (20 mg PbAc/kg bwt); PbAc +VC-treated group (20 mg PbAc/kg bwt plus 20 mg
Keywords: VC/kg bwt); and VC-treated group (20 mg VC/kg bwt). The Experimental period was lasted for 60
Lead-acetate successive days in which PbAc was administered once daily while VC was supplemented every other
Vitamin C day using intra-gastric intubation. At the end of the experimental period, all rats were sacrificed and
Male rat pathological examinations were performed. Control and VC-supplemented rats showed normal liver,
Toxicity
kidney, brain, and testes histology. In contrast, the liver of PbAc-intoxicated rats exhibited degenerated
Histopathology
hepatocytes and portal inflammatory cell infiltrations. The kidneys showed degenerated glomeruli and
formation of karyomegalic cells containing intranuclear inclusions in the proximal tubular epithelium.
Cerebellar edema, cerebral satellitosis and encephalomalacia observed in the brain. Testicular tissues
showed arrest of spermatogenesis and interstitial edema. Co-administration of VC with PbAc
diminished the severity of pathological changes and reduced the number of affected organs compared
to PbAc-intoxicated rats. These results show that low level of VC ameliorated and mitigated the adverse
pathological impacts of chronic lead toxicity.
& 2009 Elsevier GmbH. All rights reserved.

1. Introduction
Lead (Pb) toxicity is probably the most common form of heavy
metal intoxication. It is well-documented as one of the most
dangerous and insidious poisons. Its continuous environmental and
occupational exposure may contribute to renal, nervous, hepatic,
hematological and reproductive disorders in man and animals
(Flora et al., 2006; El-Sayed and El-Neweshy, 2009, Ashry et al.,
2010). The absorbed Pb is conjugated in the liver and passed to the
kidney, where a small quantity is excreted in urine and the rest
accumulates in various body organs and affects many biological
activities at the molecular, cellular and intercellular levels, which
may result in morphological alterations that can remain even after
Pb levels have fallen (Jarrar, 2003; Sidhu and Nehru, 2004; Taib
et al., 2004; Flora et al., 2006). Lead-induced oxidative stress or
disruption of prooxidant/antioxidant balance in blood and other
soft tissues has been postulated to be the major mechanism of Pb-
Abbreviations: AAS, atomic absorption spectrophotometry; HE, hematoxylin and
eosin; HPF, high power field; NaAc, sodium-acetate; NIH, National Institutes of
Health; Pb, lead; PbAc, lead-acetate; ROS, reactive oxygen species; VC, vitamin C;
D-ALAD, d-amino levulinc acid dehydratase
n
Corresponding author. Tel.: + 20 101085567; fax: + 20 453591018.
E-mail address: yasser_tf@yahoo.com (Y. Said El-Sayed).
associated tissue injury (Pande et al., 2001; Flora et al., 2003). It
causes oxidative stress by inducing the generation of reactive
oxygen species (ROS) (Gurer and Ercal, 2000), increasing the level
of lipid peroxidation and thiobarbituric acid-reactive substances
(Adonaylo and Oteiza, 1999; Upasani et al., 2001; Flora et al., 2003;
Ashry et al., 2010), and inhibiting the activity of many antioxidant
enzymes, including glutathione (Sidhu and Nehru, 2004; Flora
et al., 2006; Ashry et al., 2010). Consequently, Pb alters the
antioxidant defense system of cells resulting in pathophysiological
events in various body organs. Hence, the therapeutic strategy of
strengthening the cell’s antioxidant capacity may fortify the long-
term effective treatment of Pb poisoning. This may be achieved
through exogenous supplementation of antioxidant molecules as
an alternative to chelation therapy (Gurer and Ercal, 2000; Flora
et al., 2003; Ashry et al., 2010).
Vitamin C (Ascorbic acid, VC), a known chelating agent with
non-enzymatic antioxidant features, was widely reported to have
the ability to protect cells from oxidative stress (Patra and Swarup,
2004; Palaniappan et al., 2005; Rai et al., 2009). It is the most
important free radical scavenger in extracellular fluids, trapping
radicals in the aqueous phase and protecting biomembranes from
peroxidative damage (Patra and Swarup, 2004; Rai et al., 2009). In
Pb-exposed rats, VC has the ability to chelate Pb, with similar
potency to that of EDTA. It may increase urinary elimination of Pb

0940-2993/$ - see front matter & 2009 Elsevier GmbH. All rights reserved.
doi:10.1016/j.etp.2009.12.003
222 M. Shaban El-Neweshy, Y. Said El-Sayed / Experimental and Toxicologic Pathology 63 (2011) 221–227

and reduce its hepatic and renal burden (Dawson et al., 1999;
Simon and Hudes, 1999). Furthermore, VC significantly reduced the
lipid peroxidation levels of liver and brain, while increased the
catalase activity (Patra et al., 2001; Rai et al., 2009). Its
supplementation in Pb-intoxicated rats was also associated with
serum biochemical alterations in the hematological system and
drug metabolizing enzymes. It significantly reduced blood, liver
and kidneys Pb, blood zinc protoporphyrin and free radical levels
while increasing the activity of blood d-amino levulinc acid
dehydratase (D-ALAD) (Flora and Tandon, 1986; Vij et al., 1998).
There is a lack of experimental research papers concerning the
protective effect of VC against long-term Pb toxicity at the
histopathological levels. Thus, this study aimed to investigate the
ameliorative effect of VC against chronic Pb-associated direct tissue
damage in the liver, kidneys, brain and testes of male rats by
observing histopathological alterations.
2. Materials and methods
2.3. Animals and experimental design
Forty male Wistar rats (six-weeks-old and weighing
100720 g) were obtained from the Laboratory Animal Breeding
Colony, Faculty of Agriculture, Alexandria University. The animals
were housed in plastic cages free from any source of chemical
contamination under controlled conditions with an ambient
temperature range of 2272 1C, relative humidity of 50 75% and
a 12 h light-cycle with free access to commercial food and water.
The standard laboratory diet is composed of 161 g/kg protein,
36.4 g/kg fat, 41.1 g/kg fiber and 12.1 MJ metabolizable energy,
and purchased from Damanhur Feed Co. (Behera, Egypt). Soft
wood shavings were used for bedding and changed during the
cleaning of cages on alternate days. All animals received humane
care in compliance with the Animal Care guidelines of the
National Institutes of Health (NIH), and the local committee
approved the design of the experiments.
After an acclimatization period of 10 days, the pubertal rats
were randomly assigned into 4 groups (ten rats per group):

2.1. Chemicals Group A (Control rats) was given 15 mg NaAc/kg bwt,

Lead-acetate [(C2H3O2)2Pb
3H2O] (PbAc) and sodium-acetate
[CH3COONa
3H2O] (NaAc) were obtained from Sigma Chemical
Co. (St. Louis, MO). Vitamin C (ascorbic acid) was obtained from
Pharco Pharmaceutical Company, Alexandria, Egypt. All other
chemicals and reagents used were of analytical reagent grade.
2.2. Heavy metal and vitamin preparation
A 0.5% Pb solution was prepared by dissolving 5 g of PbAc in
1000 mL of distilled acidified water to prevent the formation of Pb
precipitate. A corresponding solution of NaAc containing equiva-
lent amount of acetate was prepared and used as a vehicle for the
control. Vitamin C dissolved in saline and used freshly in
experimental animals.
intra-gastrically once daily;
Group B (PbAc-treated rats) received 20 mg PbAc/kg bwt,
intra-gastrically once daily (Institóris et al., 1999);
Group C (PbAc+VC-treated rats) received 20 mg PbAc/kg bwt,
once daily and were concurrently supplemented with 20 mg
VC/kg bwt, every other day 30 min before PbAc administration
using intra-gastric intubation;
Group D (VC-treated rats) received 15 mg NaAc/kg bwt,
intra-gastrically once daily and were concurrently supplemen-
ted with 20 mg VC/kg bwt, every other day 30 min before NaAc
administration using intra-gastric intubation.
The timing and dose of vitamin C pretreatment have been
selected on the basis of previous reports, and to build antioxidant
pool in animal body before heavy metal exposure (Rai et al., 2009;
Mor and Ozmen, 2010). The experiment was conducted for 60

Table 1
Incidence and severity of histopathological lesions in the liver, kidneys, brain and testes of lead-acetate (PbAc)-treated group (20 mg/kg bwt/24 h, orally) (group B), and
PbAc + vitamin C (VC)-treated group (20 mg PbAc/kg bwt/24 h, orally plus 20 mg VC/kg bwt/48 h, orally) (group C).

Organ/Lesion Incidence and Severity of Histopathological Lesions

Group B (PbAc-treated rats) Group C (PbAc + VC-treated rats)

Normal ( ) Mild (+ ) Moderate (++) Severe (+++ ) Normal (–) Mild (+ ) Moderate (++) Severe (+++ )

Liver
Portal inflammatory cells infiltration 0 1 7 2 4 5 1 0
Degenerated hepatic cells 0 3 6 1 5 3 2 0
Kidneys
Damaged glomeruli 0 4 3 3 4 5 1 0
Degenerated cortical tubules 0 2 3 5 1 6 1 2
Necrotic cortical tubules 3 2 2 3 9 1 0 0
Brain
Cerebellar white matter edema 2 2 3 3 9 1 0 0
Cerebrocortical necrosis(encephalomalacia) 1 2 5 2 10 0 0 0
Cerebrocortical satellitosis 0 2 7 1 4 2 4 0
Cerebrocortical neurophagia 1 3 5 1 10 0 0 0
Cerebrocortical edema 0 1 7 2 5 4 1 0
Perivascular monocytic aggregations in cerebral cortex 0 3 7 0 7 2 1 0
Testes
Aspermia 0 0 1 9 9 1 0 0
Shrunken and wavy outlined seminiferous tubules 0 0 2 8 8 2 0 0
Binucleated giant cells formation 7 3 0 0 10 0 0 0
Intertubular edema 0 0 0 10 2 2 6 0
Damaged germinal cells 0 0 1 9 4 4 2 0

n
Number of rats with lesions per total examined (10 rats per group).
 M. Shaban El-Neweshy, Y. Said El-Sayed / Experimental and Toxicologic Pathology 63 (2011) 221–227 223

successive days. The drinking water was analyzed for the

presence of Pb using graphite furnace atomic absorption spectro-
photometry (AAS) that ensured the absence of Pb.

2.4. Collection and preparation of tissue samples

At the end of the experiment, rats from each group were

anesthetized with light ether then sacrificed by decapitation.
After animal dissection, the liver, kidneys, brain and testes were
removed, thoroughly washed with a physiological saline (0.9%
NaCl) solution and blotted on filter paper. Organ specimens were
rapidly fixed in Bouin’s solution for 4 h then retained in 70%
alcohol until processing (Carson, 1992). The fixed specimens were
processed using a conventional paraffin embedding technique.
From the prepared paraffin blocks, 5 mm thick sections were
obtained and stained with hematoxylin and eosin (HE) for light
microscopic examination (Culling, 1983).
Each tissue sample was examined for the presence of
histopathological lesions. The severity of the histopathological
lesions of each organ was assessed and graded according to the
following scale: normal ( ), mild ( + ), moderate (++) and severe
(+++). In the kidney, the presence of karyomegaly and intranuclear
inclusions was scored as: abundant (+++) if alterations were 410
per high power field (HPF) (400  ), moderate (++) if 5–10 per HPF
and rare ( + ) if o5 per HPF.

3. Results

Hematoxylin and eosin stained sections of liver, kidneys, brain

and testes were evaluated under light microscopy. The number of
rats with certain types of histopathological findings and their
occurrence in PbAc-intoxicated and PbAc-intoxicated VC-supple-
mented groups are summarized in Table 1.

3.1. Liver

The non-intoxicated control animals (group A) showed normal

hepatic histology of intact portal areas and normal hepatocytes
(Fig. 1A). In PbAc-intoxicated rats (group B); the histopathological
examination of their liver tissues revealed remarkable
degenerative changes represented by diffuse disorganization of
the hepatic cords and cytoplasmic vacuolization. Also, there are
inflammatory cells infiltration within the portal areas had been
observed (Fig. 1B). On the contrary, rats treated with PbAc plus VC
showed mild portal edema with few inflammatory cells
infiltration beside focal hydropic degeneration of the adjacent
hepatocytes (Fig. 1C). VC-treated rats (group D) showed normal
hepatic histology as that of the control group.

3.2. Kidneys

The kidneys of control rats exhibited normal renal tissue,

where normal glomeruli, tubular epithelium and interstitial tissue
were observed (Fig. 2A). Lead-induced nephropathy was apparent
in PbAc-intoxicated group. Karyomegaly with eosinophilic
intranuclear inclusions was abundant ( 410 per HPF) in the
epithelial cells of proximal tubules of six rats and moderate (5–10
per HPF) in another four rats. Glomerular damage and tubular
necrosis with invading inflammatory cells were also characteristic
lesions (Figs. 2B1 and 2B2). Rats treated with PbAc plus VC
exhibited mild karyomegaly with eosinophilic intranuclear
inclusions in the epithelial cells of proximal tubules of five rats
and the other five rats were normal. Focal cloudy swelling of the
tubular epithelium with normal glomeruli was noted (Fig. 2C).
Fig. 1. Photomicrograph of rat liver stained with hematoxylin and eosin (  250):
(A) The histoarchitecture of the liver is intact in controls. (B) Rats treated with
lead-acetate (PbAc) (20 mg/kg bwt) show disorganization of hepatic cords, diffuse
cytoplasmic vacuolization beside portal inflammatory cells infiltration (blue arrow
head). (C) The hepatic parenchyma showing mild portal edema with few
inflammatory cells infiltration (blue arrow) and focal hepatic hydropic degenera-
tion (black arrow heads) in rats receiving PbAc (20 mg/kg bwt) and supplemented
with vitamin C (VC) (20 mg/kg bwt). For interpretation of the references to colour
in this figure legend, the reader is referred to the web version of this article.
224 M. Shaban El-Neweshy, Y. Said El-Sayed / Experimental and Toxicologic Pathology 63 (2011) 221–227

Fig. 2. Photomicrograph of rat renal cortex stained with hematoxylin and eosin: (A) The histoarchitecture of the renal cortex is intact in control rats (  400). (B) Rats
intoxicated with lead-acetate (PbAc) (20 mg/kg bwt) show degenerated glomeruli (black arrows). Karyomegalic cells, characterized by enlarged nuclei containing
intranuclear inclusions, are detectable among normal tubular epithelial cells (B1, red arrows,  250). Intranuclear bodies in the epithelial lining of the proximal tubules are
conspicuous (B1 inset, red arrow,  1000). Cortical renal tubules show various degenerative changes with focal tubular necrosis invaded by inflammatory cells (B2, blue
arrow head,  400). (C) A cloudy swelling of the epithelial lining of some convoluted tubules (back arrow head) is seen in rats receiving lead-acetate (20 mg/kg bwt) and
supplemented with vitamin C (VC) (20 mg/kg bwt). For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this
article.

Vitamin C-supplemented rats, like the control group, showed rats (group B), it produced varying degrees of damage to the
normal renal architecture and histology. seminiferous tubules. They were shrunken, had a wavy outline
and were widely dispersed from each others by faint eosinophilic
material called ‘‘intertubular edema’’. Aspermia was a
3.3. Brain
conspicuous in this group, and all seminiferous tubules lacked
spermatocytes and spermatids, and were lined only with
Upon histopathological examination of control rats, brain
spermatogonia. The other germinal epithelium and Sertoli cells
tissue was normal (Fig. 3A). The cerebellar white matter in the
were necrotic. Binucleated giant cells were also encountered in a
PbAc-treated group (group B) showed diffuse edema
few seminiferous tubules (Figs. 4B and 4B inset). With VC co-
characterized by variable-sized fluid-filled spaces (Fig. 3B1). In
administration (group C), there was mild intertubular edema and
contrast, the cerebellum was found to be like the control group
mild vacuolization of the germinal epithelium of few seminiferous
(group A) in PbAc-intoxicated plus VC-supplemented rats (group
tubules leading to narrowing of the lumen. Germ cell types
C) (Fig. 3C1). The cerebral cortex in PbAc-intoxicated rats
(spermatogonia, spermatocytes and spermatids) could be
exhibited encephalomalacia characterized by variable-sized
identified in most of the tubules; no multinucleated giant cells
spaces with indefinite outlines filled with cellular debris.
were detected beside intact basement membrane (Fig. 4C). In VC-
Perineuronal satellite oligodendroglias surrounding small
supplemented rats (group D), there were well-organized
degenerated neurons with condensed chromatin and little
seminiferous tubules with intact interstitial tissue, which was
cytoplasm, neurophagia and cerebrocortical edema were also
nearly identical to the control group.
noticed in the PbAc-treated group. Monocytic accumulations in
the perivascular spaces had been recruited from the vasculature.
These cells became macrophages that phagocytose cellular debris
4. Discussion
from necrotic neurons following neuronal death (Figs. 3B2 and
3B3). However, only mild perineuronal satellite oligodendroglia
There is evidence that some nutrients, especially VC, exhibit
was observed in PbAc-intoxicated rats receiving VC (Fig. 3C2).
some protective effects against Pb intoxication (El-Zayat et al.,
Vitamin C-supplemented rats (group D) exhibited normal
1996; Houston and Johnson, 2000). Herein, the beneficial role of
cerebellum and cerebrum histology.
co-administration of a low dose of VC with PbAc is shown in the
main target organs; liver, kidneys, brain and testes, and expressed
3.4. Testes as histopathological scores. It has been clearly shown that Pb
intoxication at a concentration of 20 mg/kg bwt induces signifi-
The testes of normal control rats (group A) showed well- cant tissue damage which might be attributed to its ability to
organized seminiferous tubules. Also, all stages of transformation generate ROS that induce oxidative damage in several tissues by
of the seminiferous epithelium from spermatogonia to sperma- enhancing lipid peroxidation (El-Sokkary et al., 2003; Iavicoli
tozoa could be seen in the tubules (Fig. 4A). In the PbAc-treated et al., 2003; Ashry et al., 2010). Patra et al. (2001) reported that Pb
 M. Shaban El-Neweshy, Y. Said El-Sayed / Experimental and Toxicologic Pathology 63 (2011) 221–227 225

Fig. 3. Photomicrograph of rat brain stained with hematoxylin and eosin: (A) The histoarchitecture of the cerebellum is intact in the control (  160). (B1) Cerebellar white
matter of rats treated with lead-acetate (PbAc) (20 mg/kg bwt) showing diffuse edema (black arrow,  160). (B2) Cerebral gray matter of rats treated with PbAc (20 mg/
kg bwt) showing encephalomalacia (red arrows), satellitosis (blue arrow heads) around degenerated neurons (  400). (B3) The neuropils in the affected area are vacuolated
(blue arrow), in addition to monocytic aggregation in the perivascular space (black arrow head) and satellitosis (blue arrow heads) around degenerated neurons (  400).
(C1) Cerebellum of rats receiving PbAc (20 mg/kg bwt) plus vitamin C (VC) (10 mg/kg bwt) was found to be close to normal (  160). (C2) Cerebral cortex showing mild
perineuronal satellite oligodendroglia (red arrow heads) in rats receiving PbAc (20 mg/kg bwt) in addition to VC (20 mg/kg bwt) (  400). For interpretation of the
references to colour in this figure legend, the reader is referred to the web version of this article.

exposure resulted in a significant increase in the lipid peroxide
level in the liver and brain and a decrease in the SOD and catalase
activities.
Autopsy studies of Pb exposure cases indicate that liver tissue
is the main organ affected among the soft tissues, followed by
the kidney cortex and medulla (Patrick, 2006). Lead-induced
hepatic damage with portal cellular infiltration, cholestasis
and biliary hyperplasia is well-documented (Shalan et al., 2005;
Hamir and Sullivan, 2008). In the present study, lead hepatotoxi-
city manifests itself in disorganization of the hepatic
cords, cytoplasmic vacuolization and invading of infiltrative
inflammatory cells (Pereira et al., 2001; El-Sokkary et al., 2005).
Since chronic lead toxicity affect on a range of cellular
enzymes particularly those involved in energy production and
associated with massive dilated mitochondria leading to
hydropic degeneration appear as cytoplasmic vacuolization
(Buchheim et al., 1998)). However, mild and, to a lesser extent,
moderate degrees of hydropic degeneration and mild portal
cellular infiltration were seen in 50–60% of rats exposed to
PbAc + VC.
Acute and chronic Pb exposure both cause histological lesions
in the kidney. These lesions are characterized by the presence of
intranuclear inclusion bodies, which were induced at significantly
lower concentrations than those associated with clinical toxic
manifestations (Alden and Khan, 2002). Another histological
indication of Pb toxicity in the rat kidney is the karyomegaly of
tubular cells (Payne and Saunders, 1978). Tubular, interstitial and
glomerular damage are also characteristic renal lesions due to Pb
toxicity. Tubular changes occur earlier than glomerular and
interstitial changes, including development of pathognomonic
intranuclear inclusions in the renal tubular epithelium
(Schraishuhn et al., 1992; Jarrar, 2003; McGavin and Zachary,
2007). Our data showed clear renal lesions associated with
chronic Pb toxicity and characterized by varying degrees of
damaged glomeruli, and degenerated proximal tubules in all
intoxicated rats in the form of karyomegaly and intranuclear
inclusion formation. Nearly, 70% of intoxicated rats showed
necrotic cortical tubules. Ceruti et al. (2002) showed a correlation
between histological alterations and Pb concentration. Inclusion
bodies rapidly disappear with chelation therapy (Alden and Khan,
226 M. Shaban El-Neweshy, Y. Said El-Sayed / Experimental and Toxicologic Pathology 63 (2011) 221–227
Fig. 4. Photomicrograph of rat testes stained with hematoxylin and eosin: (A)
Most seminiferous tubules are well-preserved in control rats (  400). (B) Rats
treated with lead-acetate (PbAc) (20 mg/kg bwt) exhibit an undulation of base-
ment membranes, interstitial edema (blue arrow heads) (  160), seminiferous
tubules lined with two germ cell layers, and primary and secondary spermato-
gonia (1 and 2, respectively). Other germ cells are necrotic with formation of
binucleated giant cells in the tubular lumen (black arrow, B inset,  1000). (C)
Most tubules are relatively well-preserved with mild interstitial edema (black
arrow head) in rats administered with PbAc (20 mg/kg bwt) plus vitamin C (VC)
(20 mg/kg bwt) (  250). For interpretation of the references to colour in this figure
legend, the reader is referred to the web version of this article.
2002). This may explain the improvement of histological findings
in the kidneys, especially number of cells exhibiting karyomegaly
and intranuclear inclusions, in PbAc-intoxicated rats supplemen-
ted with VC.
Lead is a well-known neurotoxin resulting in varying degrees
of edema of the brain, degeneration of white and gray matter of
both the central and peripheral nervous system (Blood and
Hinchcliff, 2000). Chronic Pb toxicity causes polioencephaloma-
lacia, laminar necrosis of the cerebrocortex and an accumulation
of macrophages (McGavin and Zachary, 2007). Sidhu and Nehru
(2004) described histological damage in the rat cerebellum and
cerebrum after 8 weeks intoxication with 50 mg/kg bwt, includ-
ing degeneration of neurons, disruption of the normal arrange-
ment of cell layers and the cells were larger with large vascular
spaces around them. Lead treatment for 2 moths was also enough
to disrupt the normal arrangement of the cellular layers of the
cerebellum. Large spaces between the Purkinje cell layer and
the granular layer were seen. Herein, the histological portrait of
the brain in PbAc-intoxicated rats showed clear structural damage
of the central nervous system, characterized by edema of
cerebellar white matter, neuronal degeneration and polioence-
phalomalacia of the cerebral cortex. This may be attributed to
oxidative damage associated with chronic Pb intoxication in the
rat brain (Adonaylo et al., 1999). In contrast, rats treated with
PbAc plus VC exhibited mild neuronal degeneration with scant
tissue reactions. Neither cerebellar edema nor cerebral encepha-
lomalacia were noted.
Chronic Pb intoxication causes significant testicular degenera-
tion, a reduction in the size of seminiferous tubules lined with few
germinal layers, degenerated spermatocytes and spermatids and
intertubular edema, which may eventually lead to complete arrest
of spermatogenesis and aspermia (Hamir and Sullivan, 2008;
Almansour, 2009, El-Sayed and El-Neweshy, 2009). Koizumi and
Li (1992) indicated that testes are more vulnerable to oxidative
stress due to their higher polyunsaturated fatty acid content.
Severe structural damage of the testicular tissue was evident and
resulted in aspermia in all PbAc-intoxicated rats. Conversely, good
testicular histology with normal spermatogenesis, except for mild
intertubular edema in some cases, was observed in PbAc-
intoxicated rats supplemented with VC.
In conclusion, it is assumed that combined supplementation
with VC ameliorated and protected rats from structural damage
due to chronic Pb toxicity by reducing the ability of Pb to interact
with critical biomolecules (Hsu and Guo, 2002), reducing the
intestinal absorption of Pb (Dawson et al., 1999) and increasing
antioxidant efficacy to inhibit lipid peroxidation enhanced by Pb
(Upasani et al., 2001).
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