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.