الفهرس 
Introduction and Review of LiteraturOnly 14 pages are availabe for public view
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Abstract
Chemotherapy is the main treatment of cancer. Cyclophosphamide is a
nitrogen mustard alkylating agent, an anticancer, anti-neoplastic agents and
immunosuppressive agent widely used in cytotoxic chemotherapy, which
disturbs DNA synthesis and cell division.
Medical plants are widely used against various diseases. Fennel oil is the
essential oil obtained from the dried ripe fruits of foeniculum vulgare. The chief
component of fennel, anethole, had different therapeutic effects.
The present study examined the histological, histochemical,
immunohistochemical and biochemical effects of cyclophosphamide on liver
and testis of rat and the ameliorative effect of fennel oil.
Animal groups:
Animals were divided into four groups
The first group:
Animals of this group (12 rats) were kept as control groups. They were
given standard diet and tap water only.
The second group:
Animals of this group (12 rats) were orally given fennel oil (1ml/kg body
weight) once a week for six weeks.
The third group:
Animals of this group (12 rats) were orally given cyclophosphamide
(15mg/kg body weight) once a week for six weeks.
The fourth group:
Animals of this group (12 rats) were orally given cyclophosphamide
(15mg/kg body weight) followed by fennel oil (1ml/kg body weight) once a
week for six weeks.
After three and six weeks, six animals from each group were sacrificed.
Liver and testis were immediately removed from rats after dissection and fixed
in alcoholic Bouin’s fluid for histopathological study and general carbohydrates
and in 10% neutral formalin for histochemical demonistration of total proteins
and for immunohistochemical demonstration of caspase-3 and Ki-67 staining
cells. Paraffin sections (5 microns thickness) were prepared and stained with
Ehrlich’s hematoxylin and counter stained with eosin for histological
examination.
For histochemical purposes, total polysaccharides were demonstrated by
periodic acid schiff’s (PAS) technique, total proteins were demonstrated by
mercury bromophenol blue method. For immunohistochemical study Avidin
biotin method was used for detection of expression of caspase-3 and Ki-67
staining cell in liver and testicular tissue.
For biochemical analysis, blood samples were collected in clean
centrifuge tubes and left to clot in room temperature and then serum separated
by centrifugation at 3000 rpm for 20 minutes. The collected serum stored at -
20°C until analysis, The activities of ALT, AST, CAT and SOD were
determined. Also the level of MDA and GSH were measured. Testosterone and
luteinizing hormone were also determined.
The results of this study showed that:
Animals treated with fennel oil showed no changes in histological,
histochemical, immunohistochemical and biochemical investigations in both
liver and testis.
I. Liver:
The histological observations of rats treated with cyclophosphamide
showed less in the normal structure of hepatic lobules. Congestion of portal
vein, leucocytic infiltration, cytoplasmic vaculation and fatty infiltration were
detected. While, the light microscopic examination in groups treated with
cyclophosphamide followed by fennel oil showed marked improvement in the
histological structure of liver.
The histochemical investigations involved the determination of total
polysaccharides and proteins content in liver. Gradual depletion in total
polysaccharides and proteins contents were observed in liver of rats treated with
cyclophosphamide. On the other hand, the restoration of total polysaccharides
and proteins contents were observed in hepatic tissues of rats which treated with
cyclophosphamide followed by fennel oil.
The immunohistochemical investigations include the expression of
Proliferating marker Ki-67 and caspase-3. The expression of Ki-67 and caspase-
3 were strong in liver of rats treated with cyclophosphamide only. While the
administration of cyclophosphamide followed by fennel oil showed slight or
weak expression of Ki-67 and caspase-3 which were nearly similar to control
group.
II. Testis
Animals treated with cyclophosphamide showed more pronounced
histological change during different periods. Exfoliated germ cells, vaculation,
degeneration and congestion were observed. On the other hand, an improvement
observed in testis sections of rats treated with cyclophosphamide followed by
fennel oil.
The content of total polysaccharides and proteins were determined by
many histochemical techniques. Treating animals with cyclophosphamide only
showed gradual depletion in total polysaccharides and proteins in testis. On the
other hand, rats treated with cyclophosphamide followed by fennel oil showed
restoration of total polysaccharides and proteins contents in testis.
The immunohistochemical investigations involved the expression of
proliferating marker Ki-67 and caspase-3. A strong expression of caspase-3 was
observed but weak expression of Ki-67 was observed in testis of rats treated
with cyclophosphamide only. While rats treated with cyclophosphamide
followed by fennel oil showed a slight expression of caspase-3 and strong
expression of Ki-67 similar to control group.
III. Biochemical analysis
In the present work, the administration of cyclophosphamide included
elevation in the activities of ALT, AST and MDA and reduction in the activities
of antioxidant enzymes CAT and SOD and decrease in the level of testosterone
and LH hormones and non-enzymatic antioxidant GSH in sera of treated rats.
On the other hand, the biochemical results of rats treated with
cyclophosphamide followed by fennel oil showed significant reduction in the activities of ALT, AST and MDA, while recorded an increase in the activities of
SOD, CAT, levels of testosterone and LH hormones and GSH when compared
with animals treated with cyclophosphamide only.
In conclusion, fennel oil has ameliorative effect against
cyclophosphamide toxicity in both liver and testis of rats.