Clinicopathological studies on the ameliorative effect of Nigella sativa oil against genotoxicity and oxidative damage in lead intoxicated rats دراسات باثولوجية اكلينيكية علي التأثير الملطف لزيت حبة البركة ضد التغيرات السمية الوراثية و التلف التأكسدي في الفئران المسممة بالرصاص.

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.

Introduction Introduction

Poisonous substances in the environment (xenobiotics) are numerous and have toxic and carcinogenic properties that cause many disease problems Lead is a common environmental and industrial pollutant that has been detected in all phases of the environment and biologic systems It belongs to the group of most toxic heavy metals in the atmosphere It belongs to the group of most toxic heavy metals in the atmosphere

Lead causes a wide range of toxic-biochemical effects in man and animals Lead causes a wide range of toxic-biochemical effects in man and animals Liver Brain Kidneys Target organs

Mechanism of action Several mechanisms have been suggested to explain how lead induce its toxic effect Oxidative stress is considered as one of the important mechanisms of toxic effects of lead. Lead causes oxidative damage through – --inducing the generation of reactive oxygen species (ROS) -- reducing the antioxidant defense systems

Plant antioxidant Recent investigations have shown that the antioxidant properties of plants could be correlated with oxidative stress defense Nigella sativa is a one of the native herbaceous plant belonging to family Ranunculacea It is widely distributed in Egypt and other Arabian countries.

Aim of the work

The present investigation was carried out to Evaluate the effect of Nigella sativa oil Evaluate the effect of Nigella sativa oil

Biochemical Serum changes Genotoxicity Oxidative stress Histopathological alterations

Materials and Methods

Experimental design

(60) Rats G1 Control D.W G2 N.Sativa N G3 Lead L G4 N….L G5 N+L G6 L….N

Time (weeks) Groups CCCCCCCCCCGp1 ──NNNNNN──G.2 ──LLLLLL──G.3 ──LLLLLLNNG.4 ──L+N ──G.5 NNLLLLLL──G.6

Sampling

1-Bone marrow samples Collected from both femurs For micronucleus test.

2- Serum samples Serum biochemical analysis ALT,AST,ALPCholes.,TP.,ALB.,GLOBBUN,CRTPhosphorus,Calcium

Liver Brain Kidneys 3- Tissues specimens Histopathological alterations Oxidative stress biomarkers TBARS GHS CAT

Results

Genotoxicity Micronucleus test

(%) Mean ± S.E. (/ 1000) Total No. (MPCE) PCE Screaned Groups ± 0.09 c GP ± 0.10 c GP ± 1.20 a GP ± 0.09 ab GP ± 1.20 b GP ± 2.09 a GP6 Effect of Nigella sativa oil on the frequency of MPCE in bone marrow cells of lead intoxicated rats.

Biochemical results

Serum biochemical analysis

Oxidative stress biomarkers

Histopathological alterations

Liver of rats gr.3(lead acetate treated group), showing several hepatic changes in the form of; pleomorphism,binucleation, vacuolation,and karyolysis. (H&E X200). Liver of rats gr.3(lead acetate treated group), showing several hepatic changes in the form of; pleomorphism,binucleation, vacuolation,and karyolysis. (H&E X200).

Liver of rats gr.4(pre-treated group of lead with N.sativa), showing thrombus in the blood vessel consisting of fibrin network with trapped leucocytes.(H&EX200). Liver of rats gr.4(pre-treated group of lead with N.sativa), showing thrombus in the blood vessel consisting of fibrin network with trapped leucocytes.(H&EX200).

Liver of rats gr.5(concurrent treated group with lead and N.sativa), showing apparently normal hepatic tissue.(H&EX200). Liver of rats gr.5(concurrent treated group with lead and N.sativa), showing apparently normal hepatic tissue.(H&EX200).

Liver of rats gr.6(post-treated group of lead with N.sativa), showing focal area of coagulative necrosis infiltrated with mononuclear cells.(H&EX200). Liver of rats gr.6(post-treated group of lead with N.sativa), showing focal area of coagulative necrosis infiltrated with mononuclear cells.(H&EX200).

Kidnys of rats gr.3(lead acetate treated group), Showing complete absense of the glomerular tufts and vacuolation of the cytoplasm of most tubular epithelium. (H&EX200). Kidnys of rats gr.3(lead acetate treated group), Showing complete absense of the glomerular tufts and vacuolation of the cytoplasm of most tubular epithelium. (H&EX200).

Kidnys of rats gr.3(lead acetate treated group), Showing cytomegaly and karyomegaly of proximal tubular cells.(H&EX400). Kidnys of rats gr.3(lead acetate treated group), Showing cytomegaly and karyomegaly of proximal tubular cells.(H&EX400).

Kidnys of rats gr.4(pre-treated group of lead with N.sativa), Showing Vacuolation of the glomerular tufts and the cytoplasm of most tubular epithelia, together with congestion in both peritubular and glomerular capillaries.(H&EX200). Kidnys of rats gr.4(pre-treated group of lead with N.sativa), Showing Vacuolation of the glomerular tufts and the cytoplasm of most tubular epithelia, together with congestion in both peritubular and glomerular capillaries.(H&EX200).

Kidnys of rats gr.5(concurrent treated group with lead and N.sativa), Showing slight edema under the bowman capsule,with slight congestion.(H&EX200). Kidnys of rats gr.5(concurrent treated group with lead and N.sativa), Showing slight edema under the bowman capsule,with slight congestion.(H&EX200).

Kidnys of rats gr.6(post-treated group of lead with N.sativa), Showing dilatation in the glomerular space,and shrinked glomerular tufts.(H&EX200). Kidnys of rats gr.6(post-treated group of lead with N.sativa), Showing dilatation in the glomerular space,and shrinked glomerular tufts.(H&EX200).

cerebrum of rats gr.3(lead acetate treated group), Showing degenerated cerebrum of rats gr.3(lead acetate treated group), Showing degenerated neurons, satellitosis, and neuronophagia.(H&EX200). neurons, satellitosis, and neuronophagia.(H&EX200).

cerebrum of rats gr.4&6(pre&post-treated groups of lead with N.sativa), Showing diffuse perivascular and cellular edema.(H&EX200). cerebrum of rats gr.4&6(pre&post-treated groups of lead with N.sativa), Showing diffuse perivascular and cellular edema.(H&EX200).

cerebrum of rats gr.5(concurrent treated group with lead and N.sativa), Showing focal area of hemorrhage,with mild cellular edema.(H&EX200). cerebrum of rats gr.5(concurrent treated group with lead and N.sativa), Showing focal area of hemorrhage,with mild cellular edema.(H&EX200).

Conclusion

lead acetate possess a genotoxic effect, causes biochemical and histopathological alterations in the liver, kidneys and brain of rats. It is plausible that these alterations are due to the impaired oxidant-antioxidant balance and enhanced oxidative stress. Supplementation of N.sativa oil (concurrent) is beneficial in ameliorating its toxic effects.