R Structural changes in the renal cortex of male albino rat following

MJMR, Vol. 24, No. 1, 2013, pages (1-6).
Saber et al
Research Article
PROTECTIVE EFFECT OF NIGELLA SATIVA OIL ON ACUTE
CADMIUM NEPHROTOXICITY IN THE RAT RENAL CORTICAL
TISSUE: HISTOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY
Entesar Ali Saber, Seham abdel Raouf abdel Aleem, Azza Hussein Ali,
Nashwa Fathy El-Tahawy and Sara Mohammed Naguib
Department of Histology, El-Minia Faculty of Medicine
Abstract
Forty-five adult (about150-200 g) male albino rats were used in this study. They were divided into 3
groups: Group I, control group; 15 rats received single buffer saline (2 ml/ kg/d) subcutaneously. Group
II, cadmium treated group; 15 rats received daily cadmium chloride (0.49 mg/kg/d) subcutaneously.
Group III; cadmium and Nigella sativa treated group;15 rats, received a daily cadmium chloride
subcutaneously in combined form with Nigella sativa oil. After 7 days the kidney tissues were taken out
and processed for immunocytochemical, light and electron microscopic study. The current study showed
that the kidneys of group II showed some degree of degeneration in the proximal (PCT) and distal
(DCT) convoluted tubules. The renal glomeruli, PCT and DCT cells showed occasional fine vacuolar
changes. More numerous mitochondria, lysosomes, and dense irregular apical microvilli were observed
by the electron microscopy. Amelioration in the histological changes in the glomeruli as well as in the
PCT and DCT was clearly observed in group III which received Nigella sativa oil administration (group
III) if compared with group II. Preserved cortical architecture was observed. There were also less
obvious degeneration and vacuolation of renal corpuscles, PCT and DCT. The tubular cells appeared
with apparent normal basal infolding, elongated mitochondria, lysosomes and basement membranes.
Conclusion: This study provides evidence that Nigella sativa oil could exert a protective effect against
cadmium induced injury by its anti-inflammatory, anti-apoptotic, and antioxidant effects.
Key words: Nigella Sativa Oil and Immunocytochemical
INTRODUCTION:
Cadmium is involved in environmental
pollution as it is widely used in paints, plastic
manufacturing, electrolysis, and industry (1). It
accumulates in various tissues after chronic
exposure with food and water(2). Tobacco
contains significant amounts of cadmium and
smoking is one of the sources of cadmium exposure in the general population(3). Atmospheric
deposition of airborne cadmium, and the
application of cadmium-containing fertilizers
and sewage sludge on farm land may lead to
contamination of soils and increased cadmium
uptake by crops and vegetables consumed by
human beings(4). With a long biological half-life
(15-30 years), toxic effects of cadmium depend
on the dose, the length of exposure, and whether
it is breathed in or ingested(5). Cadmium
absorption from the gastrointestinal tract is the
main route of its entry(6). Cadmium causes liver,
kidney, and testis degeneration; hypertension;
132
atherosclerosis; osteoporosis; anemia; and
cancer(7). Przialeck et al.,(1) observed the effects
of cadmium on vascular permeability, edema,
hemorrhage, and necrosis. Cadmium exposure
results in necrotic and apoptotic cell death(8).
The PCT has been reported to be the primary
site of damage(9). It can also affect distal tubules
and glomerular structures(10). In the Middle East,
Northern Africa, Far East and Asia, Nigella
sativa has been used traditionally for entries for
the treatment of various diseases for over 2000
years(11). Nigella sativa contains 36-38% fixed
oil, proteins, alkaloids, saponins and 0.4-2.5%
essential oil(12). The main compounds are
thymoquione (30-48), p-cymene (7-15%), carvacrol (6-12), 4-terpineol (2-7%), t-anethole(14%) and the sesguiterpene long-ifolene (18%)(13). Many therapeutic effects of Nigella
sativa extracts have been documented, including
antioxidant(13), antimicrobial and antihelminitic(14), anti-inflammatory(15), anti-tumour(16),
Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
antidiabetic(17) and antiulcerative effects(18) in
both clinical and experimental studies.
The aim of this study was to describe the
structural and ultrastructural changes in the
renal cortex of adult male rats after acute
cadmium administration and shed a light on the
possible protective effect of Nigella sativa oil
against cadmium induced nephrotoxicity.
Material and Methods
Animals: This study was conducted in the
Histology department of faculty of Medicine of
El-Minia University. The study was dealing
with the kidney tissues of adult albino rats. A
total number of forty five male albino rats
weighing 150-200 g were used. The rats were
housed in plastic cages (as 5 rats /cage) and
maintained in a light, humidity and temperature
controlled environment for one week prior to
the experiment. Standard rat diet and water were
allowed kept on normal diet and tap water ad
libitum.
Reagents: Cadmium chloride powder was
obtained from Sigma Aldrich (Sigma Aldrich,
Egypt), which was freshly dissolved in isotonic
NaCl according to manufacturer’s instructions.
Nigella sativa oil was purchased from Origo
Company (Origo, Egypt). The company
produced Nigella sativa oil by cold pressing of
fresh seeds without using any chemicals.
Experimental design: Animals were randomly
divided into three equal groups as following:
1. The control group (group I): It consists of
15 rats which received isotonic NaCl (2ml/
kg/d) subcutaneously throughout the
experiment.
2. The cadmium treated group (group II): It
consists of 15 rats in which cadmium
chloride was given in a dose of (0.49 mg/
kg/d) subcutaneously according to Kanter et
al.,(19).
3. The cadmium and Nigella sativa treated
group (group III): It consists of 15 rats.
Rats were received a daily cadmium
chloride (0.49mg /kg/d) subcutaneously in
combined form with Nigella sativa oil in a
dose of (2 ml /kg/d) orally according to
Ebru et al.,(20)
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Saber et al
The rats were sacrificed on the 7th day by
decapitation under brief halothane anesthesia.
The kidneys were rapidly removed, divided into
two halves for separate processing. The first
half processed for light microscopy (using H &
E) and immunostaining (COX2) and the second
half processed for transmission electron
microscopy.
Image capture: Tissue sections were examined
and images were digitally captured using a
hardware consisting of a high-resolution color
digital camera mounted on an Olympus microscope (Olympus CH30, Japan), connected to a
computer, and then analyzed using Adobe
Photoshop.
Results
1) control group:
Histological study of the rat renal cortical
tissue by H & E showed numerous renal
corpuscles, proximal (PCT) and distal convoluted tubules (DCT) (fig. 1A). The renal corpuscles contained tufts of capillaries, the
glomeruli which surrounded by the Bowman's
capsules. The urinary spaces were present
between the glomeruli and Bowman's capsules.
The parietal and visceral layers of each
Bowman's capsule were distinct. The parietal
layers were lined by simple squamous epithetlium. The glomeruli were in intimate contact
with the visceral layer of Bowman's capsules
which was composed of modified epithelial
cells called podocytes. The PCT were lined
with typical thick large cubical epithelium with
acidophilic cytoplasm. The DCT showed
considerably lower cubical epithelium surrounding a relatively larger regular distinct lumens
with less acidophilic cytoplasm than the cells
of the PCT (fig. 3A). The DCT showed modification called the macula densa. The cells of
the macula densa were tall, narrow, and pale
with centrally placed nuclei. Because of the
narrowness of these cells, the densely staining
nuclei were near to each other; collectively,
viewed with the light microscope as a dense
spot (figs. 2A, 3A).
Regarding immunocytochemical study using
COX2, the renal cortex showed faint cytoplasmic expression in few macula densa cells
Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
while the cells of PCT and DCT displayed
negative COX2 immunoreactivity (figs. 5A &
6A).
At the ultrasructural level, the endothelial
cells of the glomerular capillaries were very
richly fenestrated with large pores which
appeared to lack any trace of a closing
diaphragm. Podocytes cells adhering to their
walls possessed short extensions in the form of
many primary and secondary processes. The
secondary processes were in contact with the
basal lamina of the capillaries walls. The foot
processes contained few or no organelles. The
filtration slits were clearly evident (fig. 7A).
The cells of PCT of control rat were high
cuboidal in shape with central rounded nuclei
with prominent nucleoli containing marginally
arranged chromatin material. The cell membranes were characterized by indistinct lateral
borders, many long apical microvilli and basal
infolding. Elongated mitochondria with intact
cristae, situated mainly between the nuclei and
the uniformly thickened basal cell membranes.
The cytoplasm showed also occasional
lysosomes (fig. 8A).
Additionally the cells of DCT were pale and
low cuboidal in shape with central nuclei with
prominent nucleoli. These cells were characterized by indistinct microvilli, clear cell
boundary and basal infolding. Elongated mitochondria within the cytoplasm between the
basal infolding were notice (fig. 9A).
2) One week cadmium treated group:
The histological changes caused by cadmium
were relatively minor in this group. The
changes were varied considerably from one
tubule to another or from one individual cell to
another in a single tubule. Mild intertubular
hemorrhages and peritubular capillary dilation
were noticed as shown in figure (fig. 1B).
Evidence of degeneration of some PCT and
DCT cells was clear. Intraluminal cellular
debris was also observed (fig. 2.B). The renal
glomeruli, PCT and DCT cells showed
occasional fine vacuolar changes (fig. 3B).
some Inflammatory cell infiltration was also
observed (fig. 4B).
More obvious cytoplasmic expression of COX2
in macula densa cells was observed (fig. 5B).
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Saber et al
Few cells of PCT and DCT started to show
cytoplasmic expression of COX2 (fig. 6B).
The previous results had been verified by
electron microscopic study. It was noticed that
the glomerular endothelial cells loss their
normal fenestrae organization. Widening of
some pedicles was clearly noticed. The
glomeruli appeared with normal filtration slits
but with mild thickening of their basement
membrane (Fig. 7B).
Furthermore, it was showed that the nuclei of
PCT cells displayed little chromatin condensation. The microvilli were dense and irregular
if compared with the control group. Loss of
basal infolding was also noticed. The
cytoplasm showed changes in the form of
appearance of vesicular mitochondria, more
lysosomes and vacuolation. Mild thickening of
the cellular basement membrane was showed
(fig. 8B).
The DCT cells showed some chromatin
condensation if compared with the control
group. Some cells showed shorter basal
infolding than that of the control group, while
other cells showed complete absence of their
basal infolding. The mitochondria may be
either elongated or vesicular. More lysosomes,
cytoplasmic vacuolation and mild thickening
of the cellular basement membrane was also
observed (fig. 9B).
3) One week cadmium and Nigella sativa oil
treated groups:
The damaging effects of cadmium in this group
were very minimal if compared with that of the
corresponding one week cadmium treated
group. Preserved cortical architecture with less
peritubular capillary dilatation were observed
as shown in figures (1C). The renal glomeruli
and convoluted tubular cells (proximal &
distal) showed less obvious degeneration and
cytoplasmic vacuolation (figs. 2C, 3C). In
some sections, there was less numerous inflammatory cell infiltration around the involved
tubules if compared with one week cadmium
treated rat (fig.4B).
It was clearly found that, the cytoplasmic
expression of COX2 was apparent normal in
macula densa cells. The PCT and DCT showed
less COX2 immunoactivity than the corres-
Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
Saber et al
ponding previous one week cadmium treated
group (figs. 5C, 6C).
At ultrastructure level, the glomeruli appeared
with apparent normal filtration slits and
basement membranes but with irregular fenestrae and few wide pedicles (fig. 7C).
The PCT cells in this group, showed little
nuclear chromatin condensation. Retained
basal infolding with elongated mitochondria
were also observed. The cells showed also
A
B
apparent normal long microvilli, lysosomes
and basement membranes but with few
cytoplasmic vacuolation (fig. 8C).
Additionally the DCT cells showed little
chromatin condensation. The cells appeared
with apparent normal basal infolding and
normal elongated mitochondria. The cytoplasm exhibited few vacuolation. Apparent
normal lysosomal content and basement membrane were also noticed (fig. 9C).
C
Fig. 1: Photomicrograph of renal cortex of: A) a control rat showing normal lobular organization of
the renal cortex. B) 1 week cadmium treated rat showing dilated peritubular capillaries (d) and mild
inter tubular hemorrhages (h). C) 1 week cadmium and Nigella sativa oil treated rat showing less
evident dilatation of peritubular capillaries (d) and hemorrhage (h) in comparison with the previous
corresponding treated group. (H&E X100).
.
A
B
C
Fig. 2: Photomicrograph of renal cortex of A): a control rat showing normal structure of renal
corpuscles (RC), PCT (p) and DCT (d). Notice the macula densa cells of the DCT (m). B) 1 week
cadmium treated rat showing the degeneration of PCT(p) and DCT (d) tubules. Notice intraluminal
epithelial cell debris (arrow). C) 1 week cadmium and Nigella sativa oil treated rat showing less
obvious distortion of PCT (p) and DCT (d) tubules in comparison with the previous treated group
(H&E X400).
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Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
Saber et al
B
A
C
Fig. 3: Photomicrograph of renal cortex of: A) a control rat showing a renal corpuscle (RC),
Bowman's space (BC) and podocytes (arrow). The PCT lined with typical thick cubic epithelium (p)
while DCT show a considerably lower cubic epithelium (d). The convoluted tubules have a relatively
regular distinct lumen. Notice the macula densa cells (m) . B) 1 week cadmium treated rat showing
fine cytoplasmic vacuoles of the renal corpuscle and evidence of vacuolation of PCT (pv) and DCT
(dv). C) 1 week cadmium and Nigella sativa oil treated rat showing less numerous cytoplasmic
vacuolation of the renal corpuscles (v), PCT (pv) and DCT (dv) cells in comparison with the previous
group (H&E X1000).
A
B
Fig. 4: Photomicrograph of renal cortex of: A) 1 week cadmium treated rat showing some
inflammatory cell infiltrating the renal parenchyma (circles). B)1 week cadmium and Nigella sativa
oil treated rat showing apparent less inflammatory cell infiltration (circle). (H&E X400).
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MJMR, Vol. 24, No. 1, 2013, pages (1-6).
A
Saber et al
B
C
Fig. 5: Photomicrograph of renal cortex of: A) a control rat showing faint COX2 cytoplasmic expression
in macula densa cells (m). B) 1 week cadmium treated rat showing COX2 cytoplasmic expression in the
PCT (green arrows) and DCT (blue arrow) cells. C) 1 week cadmium and Nigella sativa oil treated rat
showing that the COX2 cytoplasmic expression seems to be normal in macula densa cells (m).
(immunohistochemistry counter stain HX x1000).
A
B
C
Fig. 6: Photomicrograph of renal cortex of: A) a control rat showing negative immunoreactivity in the
PCT and DCT cells. B) 1 week cadmium treated rat showing COX2 cytoplasmic expression in the
PCT (green arrows) and DCT convoluted tubular cells (blue arrow). C) 1 week cadmium and Nigella
sativa oil treated rat showing less noticeable COX2 cytoplasmic expression in PCT(green arrows ) and
DCT (blue arrows) cells if compared with the corresponding group (immunohistochemistry counter
stain HX x1000).
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MJMR, Vol. 24, No. 1, 2013, pages (1-6).
A
B
Saber et al
C
Fig. 7: An electromicrocraph of the renal glomerulus of: A) a control rat showing a blood capillary
with fenestrated endothelium (black arrow) and podocyte (p) with primary (red arrow) and
secondary process (blue arrow). Notice the filtration slits (green arrow) and the glomerular basement
membrane (circle). B) 1 week cadmium treated rat showing blood capillaries with irregular
fenestrae(black arrow) and wide pedicles (blue arrows). Notice the apparent normal filtration slits
(green arrow) and the mild thickening of the glomerular basement membrane (circle). C) 1 week
cadmium and Nigella sativa oil treated rat showing blood capillaries with irregular fenestrae (black
arrow) and few wide pedicles (blue arrow). Notice the apparent normal filtration slits (green arrow)
and the glomerular basement membrane (circle). X14900.
A
B
C
Fig. 8: An electromicrocraph of the PCT cells of: A) a control rat showing rounded euchromatic
nucleus (n) with prominent nucleolus (nu). Notice the long microvilli (mv), basal infolding (f),
elongated mitochondria (m), indistinct lateral border (b), lysosomes (L) and cellular basement
membrane (circle). B) 1 week cadmium treated rat showing little chromatin condensation (n), dense
irregular microvilli (mv), vesicular mitochondria (m) and absence of their basal infolding. Notice the
more numerous lysosomes (l) and vacuolation (v). Cellular basement membrane becomes more
thickened (circle). C) 1 week cadmium and Nigella treated rat showing less chromatin condensation
(n), apparent normal microvilli (mv) and lysosomes (l) but with few vacuolation(v). Retained basal
infolding (f) with elongated mitochondria (m) are noticed. Notice the apparent normal thickening of
their basement membrane (circle). X7450.
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MJMR, Vol. 24, No. 1, 2013, pages (1-6).
Saber et al
.
A
B
C
Fig. 9: An electromicrocraph of the DCT cells of: A) a control rat showing rounded euchromatic
nuclei (n) with prominent nucleoli (nu). Notice indistinct microvilli, basal infolding (f) with basal
mitochondria (m), clear lateral border (b), lysosomes (L) and the cellular basement membrane (circle).
B) 1 week cadmium treated rat showing more chromatin condensation (n). The basal infolding may
appear shorter (f1) or completely absent (f2). Notice the elongated and vesicular mitochondria (m1,
m2). More lysosomes (l) and vacuolation(v) are noticeable. The cellular basement membrane
becomes more thickened (circle). C) 1 week cadmium and Nigella sativa oil treated rat showing little
chromatin condensation (n), apparent normal lysosomes (l) and basal infolding (f) with elongated
mitochondria(m). Notice the few vacuolation (v) and normal apparent basement membrane thickness
(circle). X7450.
DISCUSSION:
This study is concerned with the description of
light and electron microscopic structural
changes in the rat kidney cortex following
cadmium administration as a promising experimental model of acute nephritis. Moreover, the
changes noticed in this vital organ can be used
as warning symptoms for human health. The
main purpose of this present research was
designed to find out the nephroprotective effect
of Nigella Sativa oil on the renal cortical tissue
if concomitantly used with cadmium. Humans
environmentally exposed to cadmium,especially
smokers, are at risk of renal dysfunction (21).
There is a considerable evidence suggesting that
the kidney is a critical organ affected by
cadmium poisoning since the pathological
changes primarily observed in this organ22).
In this study, the dose of cadmium was selected
so that it was comparable with that of a human
who was occupationally exposed to high levels
of this metal(23). This dose seemed to be lower
than the amount of cadmium to which persons
living in polluted areas are exposed over a
lifetime(24).
139
This study showed various morphological
changes in the renal cortex after acute cadmium
exposure. The renal glomeruli, PCT and DCT
cells exhibited mild cytoplasmic vacuolation.
Tripathi and Srivastav(25) explained that, the
tubular vacuolezation might be an indicator of
hydrolytic changes in the renal tissue and
indicated that the toxicant might cause a failure
in the ion pump transport of tubular cells which
caused swelling of epithelium and degeneration
of tubules. These alterations also suggested
incapability of renal cells to cope with functional disturbances provoked by toxicants.
In this work, dilated peritubular capillaries and
inter tubular hemorrhages were found in some
sections. These finding were explained by
Prozialeck et al.,(1) who suggested that cadmium
increased the permeability by affecting the
communicating units between endothelia of the
venules and capillaries, as a result, edema,
hemorrhage, and necrosis occurred. It was also
in the same line of prozialeck et al.,(26) who
found that, the cadherin molecule (cellular
adhesion molecules) was decreased secondary
to the toxic effects of cadmium.
Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
In this study a few renal tubules showed single
epithelial cells desquamated to their lumen.
Tripathi and Srivastav(25) suggested that, the
presence of these amorphous substances in the
tubules might be an indication of glomerulonephritis and/or incapability of renal tubules to
counter the accumulated residues resulting from
metabolic and structural alterations caused by
cadmium.
Some inflammatory cells were evident invading
the parenchyma of the renal cortex. The
appearance of these inflammatory cells reinforced the previous recorded results which
mentioned that cadmium had been regarded as
one of the inflammation-related xenobiotics as it
induced a complex inflammatory response in
several cell types(27). Cyclooxygenase (COX) is
an enzyme that is responsible for formation of
important biological mediators called prostanoids, including prostaglandins, prostacycline
and thromboxane. COX2 is undetectable in most
normal tissue. It is an inducible enzyme in most
tissues exposed to inflammation(28). While in the
kidney, COX2 expression exhibits constitutive
expression(29). Gene targeting studies indicated
that, COX2 expression is required for the normal
development of the kidney(30). In this study
immunehistochemical detection of COX2
showed that COX2 expression was restricted to
macula densa cells in the control group. Harris
et al.,(31) previously described the same expression in the same site of the rat's distal tubular
cells. Komhoff et al.,(32), added that COX2 in rat
Macula densa cells was related to renin release.
As inflammation could play a major role in the
renal damage produced by exposure to
cadmium, one of the purposes of this study was
to assess the expression of COX2 enzymes
associated with inflammation. COX2 cytoplasmic expression in the renal cortex after one
week of cadmium administration was higher
than in control rats, not only restricted to macula
densa cells but this expression extend to involve
PCT and DCT cells.
Ultrastructural picture in this group were in
harmony with the previous results. In this study,
the ultrastructural alternations occurring in the
kidney were an immediate response to acute
cadmium exposure with signs of cellular injury.
Both glomeruli and tubules seemed to be
sensitive to acute cadmium toxicity. The glome-
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Saber et al
rular ultrastructural changes included loss of
normal organization of fenestrae, widening of
some pedicles and loss of the homogenous
appearance of basement membranes displaying
thickening in many areas. These finding were
confirmed by another study done by Asar et
al.,(33) who stated that, Glomeruli showed
increased glomerular basement membrane
thickening and irregular foot processes on acute
cadmium exposure.
In this work the PCT and DCT cells showed
gradual focal loss of their basal infolding and
apical microvilli with time of exposure. This
was in the same line of Condron et al.,(34) who
stated that, cadmium could reduce the surface
density of microvillus membrane of convoluted
tubules per unit cell volume to 19% in cadmium
contaminated rats.
Another outstanding abnormality seen in the
tubular cells was the obvious numerous
lysosomes seen in this group. The functions of
lysosomes are not fully understood, but this
organelle is known to be concerned with the
segregation and, degradation of substances
taken up by cells from the environment, as well
as of cytoplasmic constituents(35). By histological methods, Nishizumi(36) revealed the
presence of heavy metals, possibly including
cadmium, in lysosomes. At any rate, it is
possible that the observed increase in this
organelle is connected with the sequestration
and excretion of cadmium given. The increase
in lysosomes, a result of the attempt to digest
this heavy metal or toxic substance, and this was
considered a general manifestation of injury.
The sequestration of damaged organelles in
lysosomes is a mechanism of cellular repair and
follows all types of sublethal injury(37).
Another important abnormality seen in the
tubular cells was apparent more vesicular and
swollen mitochondria. The mitochondrial
swelling observed in this study may reflect the
disturbances in oxy-reduction processes taking
place in this organelle(38).
The latter finding was confirmed by Lehninger
who explained that, the presence of cadmium in
this organelle membrane was uncertain.
However, because at least 25% of the proteins
of mitochondrial membranes were respiratory
enzymes, oxidative phosphorylation in mitocho-
Protective effect of nigella sativa oil on acute cadmium
MJMR, Vol. 24, No. 1, 2013, pages (1-6).
Saber et al
ndria could be inhibited by the presence of
cadmium ions(39). Furthermore, the swelling of
mitochondria might result from a changed
intramitochondrial redox potential or an
increased mitochondrial production of oxgen
free radical(40). Ochi et al.,(41) added that,
cadmium interacts with critical subcellular sites
such as the mitocho-ndria, peroxisomes, and
microsomes and resulted in generation of free
radicals.
arachidonic acid may result in increased
production of vasoconstrictors like thromboxane, over vasodilators (prostaglandins).(46)
Furthermore, Nigella Sativa was also reported to
produce a marked inhibition in the of leukotrieens release, which cause mucosal tissue
injury and hypoxemia.(47) Therefore, it may
favor cytoprotection. Ragheb et al.,(48) highlight
the importance of reactive oxygen species in
renal pathophysiology.
In the current study there was a considerable
amelioration in the kidney cortex damaging
effect of cadmium when Nigella Sativa oil given
concurrently with cadmium. At the light and
electron microscope level, this amelioration was
in the form of, less noticeable cytoplasmic
vacuolation and distortion in the glomerular and
tubular cells, lacking evidence of major
morphological injury, reduction in the inflammatory cell infiltration and preservation of the
general structural architecture were also
observed. These morphological findings were
in the same line with Massadeh et al.,(42) who
showed that cadmium concentration in different
mice organs was of
high percentage if
compared with that when cadmium intake was
combined with Nigella sativa. It was also in
accordance to Sultan et al.,(43) who found that,
Nigella sativa oil supplementation was more
effectual in ameliorating the multiple organ
toxicity in oxidative stressed modeling.
Ahmed and El-Mottaleb(49) added that, concomitant administration of Nigella sativa oil and
acetaminophen produced a significant normalization of physiological parameter as (Na, K,
creatinine) as well as suppression the structural
changes following acetaminophen administration.
These previous morphological findings were in
agreement with Alhibshi et al.,(44) who found
that thymioquinone which is one of most active
ingredients of Nigella sativa, inhibited the
mitochondrial membrane potential deporization
and reactive oxygen generation. Also these
finding were in accordance to Saleem et al.,(45)
who suggested that, vitamin C and Nigella
sativa oil both had nephronprtotective effect as
they lowered the values of nephrotoxicity
indicators (serum creatinine, blood urea
nitrogen, and antioxidant activity) as compare to
gentamicine treated group.
The reason for the protective action of Nigella
sativa oil is not clear, but may be related to the
antioxidant action of the extract. There is
evidence that reactive oxygen species (which
was induced by cadmium), may lead to lipid
peroxidation of cell membranes and cause
degradation of phosphor-lipids. Degradation of
141
It was clearly noticed the remarked reduction in
mitochondria in the PCT and DCT in this group.
This finding are supported by the results of
Mahmoud et al.,(50) who suggested that thymoquinone, the main constited of the volatile oil of
Nigella sativa oil, is effective in protecting mice
against nephrotoxicity possible via increased the
mitochondrial function and ATP production as
well as increased the expression of antioxidant
genes, superoside dismutase, catalase and glutathione peroxidase
In conclusion, our results demonstrated the
ameliorative effects of Nigella Sativa oil supply
on the cadmium induced toxic structural
changes in the rat renal cortical tissues. Taken
together, our study suggested that a diet rich in
natural Nigella Sativa oil or used as an herbal
medicine could be useful to prevent cadmium
induced nephrotoxicity in humans exposed to
this toxic (industrial areas or smokers) and give
a good idea for broad application of Nigella
sativa oil in clinical trial. Further investigations
on the mechanism of action of Nigella Sativa oil
are required and may have a considerable
impact on future clinical treatments of patients
with acute renal failure. Further studies are also
needed to explain cadmium Nigella sativa oil
inter action in condition of long term co
exposure and their consequences for health.
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