Camel Milk Regulates Gene Expression and Activities of Hepatic Antioxidant Enzymes in Rats Intoxicated with Carbon Tetrachloride
The objective of the present study was to investigate the effect of camel milk
on activities and gene expression of hepatic antioxidant enzymes in rats intoxicated
with carbon tetrachloride (CCl4). Therefore, twenty four rats were
used in the current study. Rats were divided into four groups, the first and
second groups were received water and camel's milk, respectively whereas rats
of both third and fourth groups were injected with CCl4 and received
water and camel's milk, respectively. After 5 weeks, liver tissues were collected
for biochemical analysis of the activities and gene expression of antioxidant
enzymes. Rats supplemented with camel milk alone showed no significant difference
in all examined parameters compared to control rats. Liver damage and oxidative
stress were evident in untreated CCl4 intoxicated rats as indicated
by significant elevation of hepatic enzymes, significant elevation in thiobarbituric
acid reactive substance (TBARS), significant reduction in reduced glutathione
level (GSH), significant reduction in the activities of catalase (CAT), superoxide
dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S transferase
(GST) and finally significant down-regulation of antioxidant enzymes gene expression
compare to control. Administration of camel milk along with CCl4
caused amelioration in CCl4-induced effects compare to the untreated
CCl4 intoxicated rats via up-regulation of antioxidant enzyme gene
expression, activation of the expressed genes and increasing the availability
of GSH. Conclusively, camel milk exerted its protective effect against CCl4
induced hepatic toxicity by modulating the extent of lipid peroxidation and
augmenting antioxidant defense system at activity and gene expression levels.
Received: September 07, 2013;
Accepted: December 20, 2013;
Published: March 04, 2014
CCl4 is one of the most toxic agent to the liver tissue and its
hepatotoxicity is due to trichloromethyl radical which produced during oxidative
stress (Stoyanovsky and Cederbaum, 1999). These radicals
stimulate the release of hepatic macrophages, kupffer cells and accompanied
increase of infiltrated neutrophils and lymphocytes (Ramadori
and Saile, 2004). The activated macrophages are released and contributed
to liver fibrosis, inflammation and injury (Canbay et
al., 2004; Saile and Ramadori, 2007) with inefficient
remedy (Lee et al., 2007). Although, progress
in the treatment of liver disease by chemical drugs has been reported, searching
for natural drug is still ongoing (Recknagel, 1983).
Currently, providing therapeutically effective natural drug for the treatment
of liver diseases is the main duty of complementary and alternative medicines.
Bioactive peptides and proteins contents of camel's milk are important for maintain
effective bioprocesses such as digestion, growth and immune responses (Yagil
et al., 1984; Korhonen and Pihlanto, 2003).
In addition, camels milk has the advantage of being stored at room temperature
for longer period than other milk (Omer and Eltinay, 2009).
The most described uses of camel's milk are reported as drug against autoimmune
diseases, dropsy, jaundice, spleenomegaly, tuberculosis, asthma, anemia, piles,
diabetes (Rao et al., 1970) and as antimicrobial
agent (El Agamy et al., 1992). In addition, camels
milk has antitoxic effect against cadmium chloride (Al-Hashem
et al., 2009; Dallak, 2009), CCl4
(Khan and Alzohairy, 2011), Cisplatin (Afifi,
2010), Paracetamol (Al-Fartosi et al., 2011),
Aluminum chloride (Al-Hashem, 2009). It has been reported
that, camel milk ameliorates CCl4 induced oxidative stress via regulation
of antioxidant enzyme activities (Al-Fartosi et al.,
2012). However, publications reported the gene expression regulation of
these enzymes by camel milk are lack so far. Therefore, the aim of the present
study was to investigate the effect of camel milk on oxidative damage and oxidative
stress related gene expression and activities in rat liver intoxicated withCCl4.
MATERIALS AND METHODS
Chemicals and kits: Paraffin oil, CCl4 (Spectrosol®
BHD chemicals ltd pool, England) and other chemicals and solvents were of highest
grade commercially available. EGTA, EDTA, sucrose, Tris, butanol, mannitol,
metaphosphoric and H2O2 were purchased from Sigma Chemical
Co. (St. Louis, MO, USA). RIPA buffer was provided by Cayman chemical company,
USA. All other chemicals were of analytical grade. Diagnostic kits for serum
alanine aminotransferase (ALT) and aspartate amino transferase (AST) were purchased
from ELIPSE, United diagnostic industry, UDI, Dammam, Saudi Arabia.
Camels milk: Camels milk samples were collected daily early
in the morning from camel farm in Camel Research Center, King Faisal University,
Al-Ahsa, Saudi Arabia. Milk was collected from camels by hand milking. The samples
were collected in sterile screw bottles and kept in cool boxes until transported
to the laboratory. The rats were given fresh milk (100 mL 24 h-1
cage-1) (Althnaian et al., 2013)
as such without any further treatment.
Animals and treatment: A total of 24 albino rats (200-250 g) were obtained from Laboratory House of College of Veterinary Medicine and Animal Resources, King Faisal University, Al-Ahsa, Saudi Arabia and acclimated for 10 days before starting the experiment. All animals were housed in standard cages (6 rats cage-1), feeding with standard laboratory diet and tap water ad libitum. The experimental animals were housed in air-conditioned rooms at 21-23°C and 60-65% of relative humidity and kept on a 12 h light/12 h dark cycle. The animals received humane care in accordance with the Guide for the Care and Use of Laboratory Animals, published by ethics of scientific research committee of King Faisal University, Saudi Arabia.
Induction of hepatotoxicity by CCl4: Liver toxicity was induced
by the intraperitoneal injection of CCl4 (1 mL kg-1 b.wt.),
1:1 diluted with paraffin oil, for two successive days of the experiment (Khan
and Alzohairy, 2011).
Experimental groups and protocol: The rats were divided randomly into
4 groups comprising 6 rats in each group and fed the same diet throughout the
experimental period. The experimental design is described as fellow:
||Rats fed only with basal diet and water and injected i/p with
Paraffin oil, this group was served as control 1 group
||Rats fed normal basal diet, injected i/p with Paraffin oil and treated
with camel's milk (100 mL 24 h-1 cage-1) (Althnaian
et al., 2013) as their sole source of drinking water, this group
was served as control 2 group
||Rats fed basal diet and water and intoxicated with CCl4 (1
mL kg-1 b.wt.), 1:1 diluted with paraffin oil on first two days
of the experiment (Khan and Alzohairy, 2011)
||Rats fed basal diet and intoxicated with CCl4 (1 mL kg-1
b.wt.), 1:1 diluted with paraffin oil on first two days of the experiment
and then treated with camel's milk (100 mL 24 h-1 cage-1)
as their sole source of drinking water
Sample collection: After 5 weeks, all rats were anesthetized with diethyl
ether. Blood samples were collected by cardiac puncture before incision of the
abdomen; 5 mL of blood samples were collected in plain tubes, serum was collected
and frozen at -30°C until the time of analysis of liver enzymes using commercial
assay kits according to the manufacture instruction. The liver tissues were
removed and liver fragments were immediately frozen in liquid nitrogen and stored
at -80°C for molecular and biochemical analysis of antioxidant enzymes.
Because of the same liver tissues of the previously published work (Althnaian
et al., 2013) were used, the methodology of liver histogram was not
mentioned in the current work.
Assessment of liver damage: Commercial diagnostic kits (United Diagnostic
Industry, UDI, Dammam, Saudi Arabia) were used for determination of ALT (EP07-500)
and AST (EP15-500) on ELIPSE full automated chemistry analyzer (Rome, Italy).
The hitopathological picture of the previous work (Althnaian
et al., 2013) was taken in consideration.
Determination of hepatic antioxidant enzymes, thiobarbituric acid reactive
substances (TBARS) and reduced glutathione: One gram of liver tissues was
homogenized in 5 mL of cold 20 mM HEPES buffer, pH 7.2, containing 1 mM EGTA,
210 mM mannitol and 70 mM sucrose. After centrifugation (1500xg/5 min) at 4°C,
the supernatant was removed and stored frozen at -80°C until the time of
analysis of superoxide dismutase (SOD). Another one gram of liver tissues was
homogenized in 5 mL of cold buffer of 50 mM potassium phosphate buffer, pH 7,
containing 1 mM EDTA. After centrifugation (10.000xg/15 min) at 4°C, the
supernatant was removed and stored frozen at -80°C until the time of analysis
of catalase (CAT), glutathione peroxidase (GPX), glutathione-S transferase (GST)
and reduced glutathione (GSH). The extent of lipid peroxidation in terms of
TBARS formation was measured by mixing one gram of liver tissues with RIPA buffer
(Item No. 10010263, Cayman chemical company, USA). After homogenization, sonication
and centrifugation (1600xg/10 min), the supernatant was removed and stored frozen
at -80°C until the time of analysis. The activities of CAT (nmol min-1
g-1 tissue; Cayman Chemical Company, USA, Catalog No. 707002), GPX
(nmol min-1 g-1 tissue; Cayman Chemical Company, USA,
Catalog No.703102), SOD (U g-1 tissue; Cayman Chemical Company, USA,
Catalog No. 706002), GST (nmol min-1 g-1 tissue; Cayman
Chemical Company, USA, Catalog No. 703302) and concentrations of GSH (μM;
Cayman Chemical Company, USA, Catalog No. 703002) and TBARS (μM; Cayman
Chemical Company, USA, Catalog No. 10009055) were determined by ELISA reader
(Absorbance Microplate Reader ELx 800TM BioTek®, USA). Results
were calculated according to the manufacture instructions.
Total RNA isolation and real time RT-PCR of hepatic antioxidant enzymes: Liver tissues (approximately 1 g of tissue per sample) were immediately added to 1 mL of TriZol reagent (Invitrogen, Carlsbad, CA) and homogenized using homogenizer (Tissue Ruptor, Qiagen GmbH, Germany). One milliliter of the tissue homogenate was transferred to a microfuge tube and total RNA was extracted by adding 0.2 mL chloroform. Afterwards, samples were vortexed vigorously for 15 sec and incubated at room temperatuer for 3 min. After centrifugation (12,000 g 15 min-1) at 4°C, the aqueous phase containing RNA was transfered into new tubes. RNA was precipitated by mixing the aqueous phase with 0.5 mL isopropyl alcohol and incubated at room temperuter for 10 min. After centrifugation at 12,000 g for 10 min at 4°C, RNA pellets were washed by mixing and vortexing with 1ml of 75% ethanol. After centrifugation (7.500 g 5 min-1) at 4°C, RNA pellets were resuspended in nuclease free water (Life Technologies. USA). The purity of RNA at 260/280 OD ratio and RNA integrity was evaluated using Multi-Mode Microplate reader (SYNERGY Mx, BIO-TEK. Winooski, Vermont, USA). Only high purity samples (OD260/280 >1.8) were subjected to further manipulation. cDNA was prepared from RNA samples according to Revers Transcription System Kit (Promega, Madison, USA) by using Bio-Rad Thermal Cycler (T100TM, Foster city, California, USA). Briefly, total RNA were activated at 70°C for 10 min and 20 μL reaction mix were made of 4 μL MgCl2, 2 μL of reverse transcription 10X buffer, 2 μL of dNTP mixture (10 mM), 0.5 μL of random primers, 0.75 μL of AMV reverse transcriptase enzyme, 1ng RNA and nuclease-free water to a final volume of 20 μL. Then the reaction was incubated at 42°C for 60 min followed by incubation at 94°C for 5 min. cDNA was diluted up to 100 μL with nuclease-free water for PCR amplification. Real time RT-PCR was performed using QuantiFastTM SYBR Green PCR Master Mix kit (QIAGEN, Hilden; Germany). The 25 μL reaction for each examined gene was prepared from 12.5 μL of master mix; 2 μL forwerd primer (10 pmol); 2 μL revers primer (10 pmol); 2 μL cDNA of the sample and 6.5 μL of nuclease-free water. Cycling parameters were, 50°C for 2 min, 95°C for 15 min, 40 cycles of 95°C for 10 sec, followed by 55°C for 30 sec and 72°C for 10 sec with final melting at 95°C for 20 sec. For each gene examined, duplicate samples from each cDNA analyzed by real time RT-PCR using the Bio-Rad CFX Manager 3.0 Software of the C1000 Touch thermal cycler-CFX96 Real time PCR(BIO-RAD, Foster city, California, USA). The â-actin mRNA fragment was used as housekeeping gene to normalize the expression data. The primer sequences are described in Table 1.
Statistical analysis: All data was presented as Mean±Standard
error of mean by using student-ttest. All tests were performed using computer
package of the statistical analysis system (SAS, 2002).
The relative gene expression of target genes in comparison to the β-actin
reference gene was calculated using the Bio-Rad CFX Manager 3.0 Software of
the C1000 Touch thermal cycler-CFX96 Real time PCR(BIO-RAD, Foster city, California,
|| Details giving primer sequences and expected product size
for the genes amplified
|SOD: Superoxide dismutase, CAT: Catalase, GPX: Glutathione
peroxidase, GST: Glutathione-S transferase
Assessment of liver function: The activities of AST and ALT were estimated
in serum samples as the liver function biomarkers. These results are given in
Table 2. The CCl4 treatment markedly affected the
liver specific enzymes. It was found that a significant (p<0.05) increase
in serum AST(130.8±01.7 U L-1) and ALT (47.4±4.0 U
L-1) activities of CCl4 treated rats compare to control
(118.9±06.0 and 29.2±0.1 U L-1), respectively. This
result suggests that these hepatic biomarkers are elevated in the serum due
to release of the enzymes from damaged liver. However a significant decrease
(p<0.05) was observed in the respective serum activities of rats given Camel
milk+CCl4 (125.6±2.0 and 34.7± 1.5 U L-1)
compared with CCl4 treated group(130.8±01.7 and 47.4±4.0
U L-1), respectively. As this work is a continuation to the previous
published work (Althnaian et al., 2013), inclusion
of histopathological picture in this study was not necessary because the same
liver samples were used. The previous study had demonstrated that, liver of
CCl4-intoxicated rats showed massive fatty change and centrilobular
necrosis in most cases. Hepatitis characterized by mononuclear cells infiltration
mostly macrophages and lymphocytes around central veins and in portal areas
was also noticed in most cases of CCl4 intoxicated rats. In addition,
the liver of CCl4-intoxicated rats and treated with camel milk exhibited
clear hepatic recovery characterized by a complete regeneration of hepatocytes
and the hepatic tissue appeared more or less normal in most cases.
Effect of treatments on hepatic lipid peroxidation: Results in Table
3 shows that the TBARS level was significantly (p<0.05) increased in
the liver of Ccl4 intoxicated rats (33.3±1.01 μM) compare
to the control (28.3±0.93 μM).
||Effect of administration of CCl4 and/or camel milk
for five weeks on serum biomarker enzymes activities of AST (U L-1)
and ALT (U L-1)
|ALT: Alanine aminotransferase, AST: Aspartate amino transferase.
Values are expressed as Mean±SE, n = 6 for each group; Significance
was calculated at p<0.05. *Significant as compared to control animals.
**significant as compared to CCl4 treated animals
||Effect of administration of CCl4 and/or camel milk
for five weeks on levels of TBARS (μM) and reduced glutathione (μM)
and activities of CAT (nmol min-1 g-1 tissue), SOD
(U g-1 tissue), GPX (nmol min-1 g-1 tissue)
and GST (nmol min-1 g-1 tissue) in rats liver tissue
|TBARS: Thiobarbituric acid reactive substance, CAT: Catalase,
SOD: Superoxide dismutase, GPX: Glutathione peroxidase, GST: Glutathione-S
transferase and GSH: Reduced glutathione. Values are expressed as Mean±SE,
n = 6 for each group. Significance was calculated at p<0.05, *significant
as compared to control animals, **significant as compared to CCl4
Treatment of CCl4 intoxicated rats with camel milk caused significant
(p<0.05) decrease in TBARS level (28.3±1.02 μM) of liver tissue
compared to that in liver from untreated CCl4 intoxicated rats (33.3±1.01
μM). Administration of camel milk only did not affect significantly (p>0.05)
the level of TABARS in rat liver compare to the control.
Effect of treatments on GSH and hepatic antioxidant enzyme activities: GSH concentration and antioxidant enzyme activities (CAT, SOD, GPX and GST) are shown in Table 3. The concentration of GSH was reduced significantly (p<0.05) in CCl4 intoxicated rats (4.0±0.10 μM) compare to the control (5.0±0.10 μM). However, camel milk administration reserves GSH values in CCl4 intoxicated rats (6.0±0.20 μM) compare to untreated CCl4 intoxicated rats (4.0±0.10 μM) and control (5.0±0.10 μM). All the antioxidant enzyme activities were reduced significantly (p<0.05) in CCl4 intoxicated rats (CAT: 23.2±1.10 nmol min-1 g-1 tissue; SOD: 6.0±0.02 U g-1 tissue; GPX: 272.1±2.10 nmol min-1 g-1 tissueand GST: 140.1±1.02 nmol min-1 g-1 tissue) compare to the control (27.0±0.20; 8.0±0.02; 290.1±0.90; 186.0±1.00), respectively. However all activities of these enzymes showed a significant (p<0.05) recovery in response to camel milk administration to CCl4 intoxicated rats (27.1±1.20; 8.0±0.01; 291.4±2.20; 188.8±2.01), respectively compare to untreated CCl4 intoxicated rats (23.2±1.10; 6.0±0.02; 272.1±2.10; 140.1±1.02), respectively. The activities of hepatic antioxidant enzymes of rats treated only with camel milk were comparable to that of the control (Table 3).
Effect of treatments on gene expression of antioxidant enzymes: The mRNA expression of SOD, CAT, GPX and GST were measured by real time PCR (Fig. 1). The results showed that, the expression of all of the antioxidant enzymes were reduced significantly (p<0.05) in CCl4 intoxicated rats compare to the control. However, the expression of all of the antioxidant enzymes were increased significantly (p<0.05) in CCl4 intoxicated rats treated with camel milk compare to untreated CCl4 intoxicated rats. The expression of hepatic antioxidant enzymes of rats treated only with camel milk was comparable to that of the control (Fig. 1).
In the current study serum liver biochemical markers, AST and ALT activities
were increased significantly (p<0.05) in rats treated with CCl4
compare to control. This increase in serum levels of liver function biomarkers
might be attributed to the liver damage as these enzymes are cytoplasmic in
location and its released into circulation indicated damage to cell membrane
(Zimmerman et al., 1965; Brent
and Rumack, 1993; Recknagel et al., 1989).
Similar results (Trible et al., 1987; Wang
et al., 1997; Mehmetcik et al., 2008;
Arici and Cetin, 2011; Althnaian
et al., 2013) demonstrated that, ALT and AST enzymes activities were
significantly increased following CCl4 administration. The hepatotoxic
effectof CCl4 is due to active metabolite Ccl3 which has
the ability to eliminate hydrogen from fatty acids and initiating the lipid
peroxidation, hepatocytes injury and liver damage (Forni
et al., 1983; Park et al., 2005).
However, in the current study, camel milk was found to ameliorate (p<0.05)
the leakage of AST and ALT induced by CCl4 treatment in rats. This
suggests a membrane stabilizing activity of camel milk because it has been accepted
that, serum levels of transaminases might be return to normal whenever healing
of hepatic parenchyma and hepatocytes regeneration have been achieved (Thabrew
et al., 1987). Supporting to the present findings, several reports
(Khan and Alzohairy, 2011; Hamad
et al., 2011; Al-Fartosi et al., 2012;
Althnaian et al., 2013) demonstrated the hepatoprotective
effect of camel milk.
||Real time PCR analysis of mRNA gene expression of hepatic
catalase, (a) CAT, (b) Superoxide dismutase, SOD, (c) Glutathione peroxidase,
GPX and (d) Glutathione-S transferase, GST of control non-treated, camel
milk treated, CCl4 treated and CCl4+camel milk treated
rats. Liver tissues were homogenized in TriZol reagent and RNA was extracted
by chloroform. After preparation of cDNA, real time RT-PCR was performed
using QuantiFastTM SYBR Green PCR Master Mix kit. The relative gene expression
of target genes in comparison to the β-actin reference gene was calculated
using the Bio-Rad CFX Manager 3.0 Software of the C1000 Touch thermal cycler-CFX96
Real time PCR. Values are expressed as mean±SEM. *Values are significantly
different (p<0.5) compare to control. #Values are significantly
different (p<0.5) compare to CCl4 intoxicated group
These studies elucidated that, the ameliorative effect of camel milk against
CCl4-induced oxidative stress in rats was attributed to the antioxidant
effect of camel milk contents such as vitamins (A, B2, C and E) and
trace elements (Yousef, 2004). The efficient hepatoprotective
drugs can be defined as, drug which able to reduce the harmful effect or restore
the normal hepatic function that has been distributed by toxin. Camel milk reduced
the levels of liver enzymes that has been elevated as results of CCl4
toxicity. This indicated that, the structural integrity of hepatocytes cell
membrane has been protected and/or damaged hepatocytes have been regenerated
(Palanivel et al., 2008).
The significant increase of TBARS in liver of untreated CCl4 intoxicated
rats suggests enhanced peroxidation leading to tissue damage and the failure
of the antioxidant mechanisms in preventing of excessive free radicals (Romero-Alvira
and Roche, 1996). This confirmed by the results of the current study which
reported a significant decrease of both GSH content and the activities of antioxidant
enzymes (SOD, GPX, CAT, GST) in CCl4 intoxicated rats compare to
control which impose in elevation of TBARS levels. These findings were found
to be in agreement with other previous studies in rats liver intoxicated with
CCl4 (Yousef, 2004; Aranda
et al., 2010; Al-Fartosi et al., 2012;
Bona et al., 2012; Pirinccioglu
et al., 2012). The significant decrease of TBARS in liver of rats
intoxicated with CCl4 and treated with camel milk suggests the protective
effect of camel milk against CCl4-induced oxidative stress. This
protective effect of camel milk is due to contents of essential vitamins and
minerals (Barbagallo et al., 1999).
GSH plays an important role in antioxidant system of the body. It maintains
normal structure and function of the cells via redox and detoxification reaction.
In the present study a significant decrease of GSH values were observed in CCl4
intoxicated rats. The decrease of GSH is perhaps due to direst requisition of
GSH by GPX to scavenge free radicals that formed during the metabolism of CCl4.
Similar results were reported in rats intoxicated with CCl4 (Bona
et al., 2012). The ameliorative effect of camel milk against CCl4
induced toxic effect on GSH level was attributed to the ability of camel milk
to scavenge free radicals and restore the antioxidant status which underlined
by increasing the activities and gene expression of antioxidant enzymes as discussed
below. Camel milk restored the GSH values in rat liver intoxicated with aluminum
chloride (Al-Hashem, 2009), cadmium chloride (Al-Hashem
et al., 2009) and alcohol (Darwish et al.,
The oxidative stress refers to the imbalance between prooxidants and antioxidants
in biological systems. Therefore, the significant increase in lipid peroxidation
could be due to significant reduction in the activities of enzymatic antioxidants
such as CAT, SOD, GPX and GST as well as non- enzymatic antioxidants such as
GSH in the liver of CCl4-intoxicated rats, as compared to the control.
Superoxide radical is converted to H2O2 by SOD. Furthermore,
H2O2 is transferred to molecular oxygen and water by CAT
and GPX. Therefore, SOD, CAT, GPX and GST constitute the principal components
of the antioxidant system and their deficiencies leads to oxidative stress.
Therefore, the significant reduction in these enzyme activities in liver of
CCl4 intoxicated rats (Table 3) could be responsible
for increased lipid peroxidation observed as reflected on high level of TBARS
along with low level of GSH in these animals compared to the control during
CCl4-induced hepatotoxicity. Significant reductions in GPX, SOD and
CAT have been reported in CCl4-intoxicated rat liver (Palanivel
et al., 2008; Bigoniya and Rana, 2010),
kidney (Adewole et al., 2007; Ganie
et al., 2011) and lung (Ganie et al., 2011).
Previous study has reported that camel milk is a potent inducer of these detoxifying
enzymes and thereby prevents the toxicity induced by aluminum chloride (Al-Hashem,
2009), cadmium chloride (Al-Hashem et al., 2009;
Dallak, 2009) and alcohol (Darwish
et al., 2012).
At gene expression level, the current findings indicated that, CCl4
induced down regulation of all examined antioxidant enzymes (SOD, CAT, GPX and
GST) gene expression. Similar results (Chen et al.,
2013) was obtained in CCl4-induced liver fibrosis in mice. The
present findings also reported that, camel milk augmented the antioxidant status
via up-regulation of CAT, SOD, GPX and GST gene expression (Fig.
1). Similar results (Afifi, 2010) demonstrated that
camel milk regulated gene expression of these antioxidant enzymes in lung tissues
of rats intoxicated with cisplain. The author studied gene expression of antioxidant
enzymes in lung tissues of rats by conventional PCR. The conjugation of reactive
xenobiotic metabolites with GSH as mentioned above is an important step in detoxification
mechanism. This conjugation process is mediated by GST. CCl4 depleted
the GSH through conjugation process (Table 3). The down regulation
of GST gene expression in CCl4 intoxicated rats compare to control
as presented in the current study (Fig. 1c) could limit the
ability of hepatic tissues to conjugate the reactive metabolites which alleviated
by Camel milk.
Camel milk may exert its protective effect against CCl4 induced hepatic toxicity by modulating the extent of lipid peroxidation and augmenting antioxidant defense system at activity and gene expression levels.
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