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Nashwa, A. Abu-Aita ; Sherein, S. A. EL Gayed ; Mogda,K.Mansoutr (2006) : Clinico pathological studies on the ameliorative effect of nigella sativa oil against genotoxicity and oxidative damage in lead intoxicated rats. J. Egypt. Vet. Med. Assoc. Vol 66 No4 ; 127–46.
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ClinicopathologicalClinicopathological studies on the ameliorative studies on the ameliorative effect of Nigella sativa oil against genotoxicity effect of Nigella sativa oil against genotoxicity
and oxidative damage in lead intoxicated ratsand oxidative damage in lead intoxicated rats
الملطف التأثير علي اكلينيكية باثولوجية دراساتالوراثية السمية التغيرات ضد البركة حبة لزيت
. بالرصاص المسممة الفئران في التأكسدي التلف و
Nashwa, A. Abu-Aita ; Sherein, S. A. EL Gayed ; Mogda,K.Mansoutr
(2006) : Clinico pathological studies on the ameliorative effect of nigella sativa oil against genotoxicity and oxidative
damage in lead intoxicated rats .J. Egypt. Vet. Med. Assoc .
Vol 66 No4 ; 127–46 .
•IntroductioIntroductionn
Poisonous substances in the Poisonous substances in the environment (environment (xenobioticsxenobiotics) are ) are numerous and have numerous and have toxictoxic and and carcinogeniccarcinogenic properties that cause properties that cause many disease problemsmany disease problems
LeadLead is a common is a common environmentalenvironmental and and industrialindustrial pollutant that has pollutant that has been detected in all phases of the been detected in all phases of the environment and biologic systemsenvironment and biologic systems
It belongs to the group ofIt belongs to the group of most most toxic heavy metalstoxic heavy metals in the in the atmosphere atmosphere
Lead causes a wide range of Lead causes a wide range of toxic-biochemicaltoxic-biochemical effects in effects in man and animalsman and animals Liver
Brain
KidneysTarget organsTarget organs
Mechanism of actionMechanism of actionSeveral mechanisms have been suggested Several mechanisms have been suggested
to explain how lead induce its toxic to explain how lead induce its toxic effecteffect
Oxidative stressOxidative stress is considered as one of the is considered as one of the important mechanisms of toxic effects of important mechanisms of toxic effects of lead. lead.
Lead causes oxidative damage through –Lead causes oxidative damage through –----inducinginducing the generation of reactive oxygen the generation of reactive oxygen
species (ROS)species (ROS)----reducingreducing the antioxidant defense systems the antioxidant defense systems
Plant antioxidantPlant antioxidantRecent investigations have shown Recent investigations have shown that the that the antioxidant properties of antioxidant properties of plantsplants could be correlated with could be correlated with oxidative stress defenseoxidative stress defenseNigella sativaNigella sativa is a one of the is a one of the native native herbaceousherbaceous plant belonging to plant belonging to family family RanunculaceaRanunculacea
It is widely distributed in Egypt and It is widely distributed in Egypt and other Arabian countries.other Arabian countries.
Aim of the Aim of the workwork
The present investigation The present investigation was carried out towas carried out to
Evaluate the effect of Evaluate the effect of Nigella sativaNigella sativa oil oil
Biochemical Serumchanges
Genotoxicity
Oxidative stress Histopathological
alterations
Materials Materials and and
MethodsMethods
Experimentaldesign
G6L….N
G5N+L
G4N….L
G3Lead
L
G2N.Sativa
N
G1Control
D.W
(60)Rats
GroupsTime (weeks)
1212345612
Gp1CCCCCCCCCC
G.2──NNNNNN──
G.3──LLLLLL──
G.4NNLLLLLL──
G.5──L+NL+NL+NL+NL+NL+N──
G.6──LLLLLLNN
Sampling
1-Bone marrow samples
Collected from both femurs
For micronucleus test.
22 - -Serum samplesSerum samples
Serum biochemicalSerum biochemical analysisanalysis
ALT,AST,ALPALT,AST,ALPCholes.,TP.,ALB.,GLOBCholes.,TP.,ALB.,GLOB
BUN,CRTBUN,CRTPhosphorus,CalciumPhosphorus,Calcium
Liver BrainKidneys
3- Tissues specimens
Histopathological alterations
Oxidative stress biomarkers
TBARSGHSCAT
ResultsResults
GenotoxicityMicronucleus test
GroupsPCEScreaned
Total No.(MPCE)
Mean ± S.E.(/ 1000)(%)
GP13000289.33 ± 0.09c0.93
GP230003210.66 ± 0.10c1.06
GP3GP330008829.33 ± 1.20a2.93
GP430008626.33 ± 0.09ab2.63
GP5GP530006521.66 ± 1.20b2.16
GP630008729.00 ± 2.09a2.90
Effect of Nigella sativa oil on the frequency of MPCE in bone marrow cells of lead intoxicated rats.
Biochemical Biochemical resultsresults
Serum biochemical analysis
S.AL T
0
5
10
15
20
25
30
35
40
45
50
6
WEEKS
IU/L
G1
G2
G3
G4
G5
G6
AST
70
75
80
85
90
95
6
WEEKS
(IU/L)
G1
G2
G3
G4
G5
G6
AL P
75
80
85
90
95
100
6
WEEKS
(IU/L)
G1
G2
G3
G4
G5
G6
CHOL E STE ROL
70
72
74
76
78
80
82
84
86
88
90
92
6
WEEKS
mg/dl
G1
G2
G3
G4
G5
G6
S.T OTAL PROTE I NS
5.5
6
6.5
7
7.5
8
6
WEEKS
g/dl
G1
G2
G3
G4
G5
G6
S.AL BU MI N
3.4
3.6
3.8
4
4.2
4.4
4.6
6
WEEKS
g/dl
G1
G2
G3
G4
G5
G6
BU N
0
5
10
15
20
25
30
35
6
WEEKS
mg/dl
G1
G2
G3
G4
G5
G6
S.CREAT I N I NE
0
0.2
0.4
0.6
0.8
1
1.2
1.4
6
WEEKS
mg/dl
G1
G2
G3
G4
G5
G6
SE RU M PHOSPHORU S
0
2
4
6
8
10
12
14
6
WEEKS
mg/dl
G1
G2
G3
G4
G5
G6
Oxidative stress biomarkers
MDA (L ive r ho moge nate ))
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
6
weeks
mM
/100
g)
G1
G2
G3
G4
G5
G6
G HS (L ive r homo ge na te )
0
50
100
150
200
250
300
350
400
450
6
Weeks
(µg/g)
G1
G2
G3
G4
G5
G6
CAT AL ASE (L ive r ho moge na te )
0
2
4
6
8
10
12
6
Weeks
(IU/m
g pr
otein)
G1
G2
G3
G4
G5
G6
Histopathologicalalterations
•Liver of rats Liver of rats gr.3(lead acetate treated group),gr.3(lead acetate treated group), showing several hepatic showing several hepatic changes in the form of; pleomorphism ,binucleation, vacuolation ,and changes in the form of; pleomorphism ,binucleation, vacuolation ,and
karyolysis. (H&E X200)karyolysis. (H&E X200) . .
•Liver of rats Liver of rats gr.4(pre-treated group of lead with N.sativagr.4(pre-treated group of lead with N.sativa), ), showing thrombus in the blood vessel consisting of fibrin showing thrombus in the blood vessel consisting of fibrin
network with trapped leucocytes.(H&EX200)network with trapped leucocytes.(H&EX200)..
•Liver of rats Liver of rats gr.5(concurrent treated group with lead and gr.5(concurrent treated group with lead and N.sativaN.sativa), showing apparently normal hepatic tissue.), showing apparently normal hepatic tissue.
(H&EX200)(H&EX200) . .
• Liver of rats Liver of rats gr.6(post-treated group of lead with gr.6(post-treated group of lead with N.sativa),N.sativa), showing focal area of coagulative necrosis showing focal area of coagulative necrosis
infiltrated with mononuclear cells.(H&EX200)infiltrated with mononuclear cells.(H&EX200) . .
•Kidnys of rats Kidnys of rats gr.3(lead acetate treated groupgr.3(lead acetate treated group), ), Showing complete absense of the glomerular tufts Showing complete absense of the glomerular tufts and vacuolation of the cytoplasm of most tubular and vacuolation of the cytoplasm of most tubular
epithelium. (H&EX200)epithelium. (H&EX200) . .
•Kidnys of rats Kidnys of rats gr.3(lead acetate treated groupgr.3(lead acetate treated group), Showing ), Showing cytomegaly and karyomegaly of proximal tubular cells.(H&EX400)cytomegaly and karyomegaly of proximal tubular cells.(H&EX400) . .
•Kidnys of rats Kidnys of rats gr.4(pre-treated group of lead with N.sativa),gr.4(pre-treated group of lead with N.sativa), Showing Vacuolation of the glomerular tufts and the Showing Vacuolation of the glomerular tufts and the cytoplasm of most tubular epithelia, together with cytoplasm of most tubular epithelia, together with
congestion in both peritubular and glomerular capillaries.congestion in both peritubular and glomerular capillaries.(H&EX200)(H&EX200) . .
Kidnys of rats Kidnys of rats gr.5(concurrent treated group with lead and gr.5(concurrent treated group with lead and N.sativa),N.sativa), Showing slight edema under the bowman Showing slight edema under the bowman
capsule ,with slight congestion.(H&EX200)capsule ,with slight congestion.(H&EX200) . .
•Kidnys of rats Kidnys of rats gr.6(post-treated group of lead with N.sativa),gr.6(post-treated group of lead with N.sativa), Showing dilatation in the glomerular space,and shrinked Showing dilatation in the glomerular space,and shrinked
glomerular tufts.(H&EX200)glomerular tufts.(H&EX200)..
•cerebrum of rats cerebrum of rats gr.3(lead acetate treated group),gr.3(lead acetate treated group), Showing degeneratedShowing degenerated
•neurons, satellitosis, and neuronophagia.(H&EX200)neurons, satellitosis, and neuronophagia.(H&EX200)..
•cerebrum of rats cerebrum of rats gr.4&6(pre&post-treated groups of lead gr.4&6(pre&post-treated groups of lead with N.sativawith N.sativa), Showing diffuse perivascular and cellular ), Showing diffuse perivascular and cellular
edema.(H&EX200)edema.(H&EX200)..
•cerebrum of rats cerebrum of rats gr.5(concurrent treated group with gr.5(concurrent treated group with lead and N.sativalead and N.sativa), Showing focal area of ), Showing focal area of
hemorrhage ,with mild cellular edema.(H&EX200)hemorrhage ,with mild cellular edema.(H&EX200) . .
Conclusion
lead acetate possess a lead acetate possess a genotoxicgenotoxic effect, causes effect, causes biochemical biochemical and and histopathologicalhistopathological alterations in the alterations in the liver, kidneys and brain of rats. liver, kidneys and brain of rats.
It is plausible that these alterations are It is plausible that these alterations are due todue to the impaired oxidant- the impaired oxidant-antioxidant balance antioxidant balance and and enhanced enhanced oxidative stress.oxidative stress.
Supplementation of Supplementation of N.sativa oilN.sativa oil
(concurrent) is beneficial in (concurrent) is beneficial in amelioratingameliorating its toxic effects. its toxic effects.