الفهرس 
Introduction & Aim of the WorkOnly 14 pages are availabe for public view
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Abstract
Summary and Conclusion Ehrlich’s tumor is a transplantable neoplasia from malignant epithelium that corresponds to mice’s mammary adenocarcinoma which can be used in experimental cancer chemotherapy studies. Today, an extremely promising strategy for cancer prevention is chemoprevention, which is defined as synthetic or natural agents and used alone or in combination to block the development of cancer in humans. Balanites aegyptiaca is a plant rich in steroids which may have a role in impairment of cancer cell proliferation.
The main objective of this work was to study the ascitic form of tumor bearing mice (Ehrlich) in its primary site (peritoneal cavity) and in two of its expected secondary sites (liver and spleen) and role of B. aegyptiaca extract in the protection.
Materials and Methods:
One hundred of adult female albino mice were used in the present study.
They were divided into three groups as following:
Group I (Control group): composed of 40 mice and were subdivided into two subgroups:
• Subgroup Ia (Plain control subgroup): composed of 20 mice that were kept on balanced diet and water adlibitum and served as control group to all groups.
• Subgroup Ib (Sham control subgroup): composed of 20 mice, received single dose of 0.2 ml of 0.9 % NaCl intraperitoneally.
Group II (Balanites aegyptiaca group): composed of 20 mice, received B. aegyptiaca fruit extract at daily dose of 500 mg/kg by intraperitoneal injection for 2 weeks.
Group III (EAC bearing group): composed of 40 mice which were inoculated intraperitoneally with a single dose of Ehrlich tumor cell line at (10)6 cells in 0.2 ml of 0.9 % NaCl/mouse for induction of tumor. The tumor growth was observed within 4-6 days following transplantation. the mice were subdivided into two subgroups:
• Subgroup IIIa (EAC control subgroup): consisted of 20 mice served as EAC control.
• Subgroup IIIb (EAC protected subgroup): composed of 20 mice, after 24 hours of induction of tumor, they received B. aegyptiaca fruit extract at daily dose of 500 mg/kg by intraperitoneal injection for 2 weeks.
At the end of the experiment ten mice from subgroup Ia, subgroup Ib, group II , subgroup IIIa & subgroup IIIb were sacrificed by cervical dislocation while the other ten mice were kept to check the MST and %ILS of EAC-tumor bearing hosts.
Before and after scarification the following data were estimated:
• Tumor growth response assessment; determined general appearance and body weight changes, ascitic tumor volume number of tumor cells, and percent of viable tumor cells.
• Clinico-pathological assessment; determined ascitic fluid levels of:
total protein, antioxidant enzymes activities; SOD & CAT enzymes and MAD. Also blood levels of AST & ALT (liver function tests),
urea & creatinine (kidney function tests).
• Cytological assessment; studied the cytology of peritoneal and ascitic fluid smears.
• Histo-pathological assessment; included the liver and spleen histological studies.
• Molecular assessment; determined p53 gene expression in the ascitic fluid of EAC bearing mice Results:
• Control and B. aegyptiaca groups (groups I & II):
In the present study, mice treated intraperitoneally with aqueous extract of B. aegyptiaca fruit showed no significant difference in the body weight, survival time, liver and kidney function tests, cytological and histopathological studies when compared to the mice of control group.
• EAC control mice (subgroupIIIa):
Comparing the EAC control mice (subgroup IIIa) to the control group(group I), it showed a decrease in mean survival time and % of increased life span and a highly significant increase in the mice body weight accompanied by huge abdominal distension. The ascitic fluid of EAC control mice (subgroup IIIa) showed a high level of total proteins and malondialdehyde (MDA); low level of SOD, CAT enzymes and p53 gene expression.
The cytological study of ascitic fluid of EAC control mice (subgroup IIIa) revealed a hypercellular smear that showed cellular pleomorphism(variable in shape and/or in size), hyperchromasia with increased nucleus/cytoplasm ratio and high mitotic count. Most of cells arranged in clusters but few of them showed linear and acinar arrangement. Some cells showed vacuolated cytoplasm and knobby borders. Both nuclei and nucleoli showed pleomorphism. Some nuclei showed lobulation with irregular dense nuclear membrane. Inbetween the malignant cells there were abundant reactive mesothelial and apoptotic cells.
Comparing the EAC control mice to normal control mice, the kidney and liver function tests showed a highly significant increase in blood urea and serum creatinine level and in AST and ALT respectively.
The histological study of the liver sections of EAC control mice supported the liver biochemical function results as it revealed parenchymal hemorrhage, necrosis and vacuolation associated with hyDROPic degeneration. Moreover, lobular shrinkage in the form of decreased the area between portal areas and central veins. Aggregated cells with malignant criteria could be detected in widened portal area.
Lastly, vascular congestion and dilatation associated with dilatation of the bile duct could be also reported.
Regarding the histological study of the spleen sections of EAC control mice (subgroup IIIa), it revealed marked decrease in follicular cellularity and periarteriolar lymphatic sheaths of the white pulp. The red pulp showed extensive vacuolations and hyalinization. Also there were capsular thickening and congested dilated splenic blood sinusoids.
EAC protected mice (subgroupIIIb):
In the present study, supplementation of B. aegyptiaca fruit extract at daily dose of 500 mg/kg for 2 weeks to EAC bearing mice (subgroup IIIb) produced significant; tumor growth response, clinico-pathological cytological, histopathological and molecular improvement when compared with EAC control mice (subgroup IIIa).
The results obtained from the present study revealed a significant decrease in the body weight, the highly significant increase in the SOD & CAT and the highly significant decrease in the MDA and total protein levels in the ascitic fluid of EAC treated mice (subgroup IIIb).
In the present study, the improvement in the cytological study of the ascitic fluid smear of EAC treated mice (subgroup IIIb) was in the form of hyocellularity, disappearance of the clusters aggregations and decrease in mitotic count. The malignant cells showed either degenerated or apoptotic figures.
In the present work, the protective effect of B. aegyptiaca fruit extract against kidney and liver were proved in the form of decrease in the levels of kidney and liver function tests. The liver showed nearly normal hepatocytes plates associated with tiny vacuoles, slight widening of portal area which housed aggregated apoptotic cells and activated sinusoidal kupffer cells. The spleen showed normal capsule and increase in the cellularity of the follicles and the periarteriolar lymphatic sheaths of the white pulp. While the red pulp showed slight vacuolations and dilatation of the blood sinusoids.
Regarding the molecular improvement, a highly significant increase of p53 gene expression in EAC treated mice (subgroup IIIb) when compared to that of EAC control mice (subgroup IIIa) was recorded.
from the present study, it could be concluded that:
1. EAC is a virulent tumor as it can:
a. Produce liver metastasis, metaplastic changes in the spleen that known as a rare site of metastasis.
b. Significantly impair the p53 function with low level of p53 gene expression as it known as tumor suppressor gene.
c. Impair the defense system in the form of immunodeficiency and increased lipid peroxidation with low level of antioxidant enzymes.
d. Cause marked decrease the hosts, survival time.
2. B. aegyptiaca could be considered as a very potent anti-cancer agent as it:
a. Inhibit the liver metastasis and metaplastic changes in the spleen and when infiltration occurs in some cases, the cancer cells undergo apoptosis.
b. Prolong the MST and %ILS of EAC bearing model to reach the level of a very potent anticancer reference drug ;
cisplatin (MST reached 29.9±1.71 days for B. aegyptiaca and 31.0±1.4 days for cisplatin)c. Reactivate p53 gene to high significant level.
3. B. aegyptiaca protect the liver from the fatty infiltration and fibrosis.
from this study it is recommended that:
• Usage of Ehrlich tumor in experimental studies should be encouraged for its virulence and metastatic effects and other organs should be inspected for metastasis.
• The cultivation of B. aegyptiaca trees should be encouraged to allow the production of cheap and effective antioxidant, antiinflammatory and anticancer medications.
• Further studies should be done using a higher dose of B. aegyptiaca and a longer duration of treatment to prevent the mild liver infiltration or even the splenic metaplastic changes.
• Further studies should be done to analyze the exact phytochemical constituents of the crude aqueous extract of B. aegyptiaca fruit mesocarp to know more about its benefits.
• Further studies should be done using a higher dose of inoculated ascitic tumor to clarify the role of B. aegyptiaca in prevention of the development of solid Ehrlich tumor at the site of tumor inoculation as well as the cancer metastasis.
• Further studies should be done to investigate the haematological reaction of B. aegyptiaca to exclude the bone marrow suppression as done by the other anticancer drugs.
• Further studies should be done to combine the B. aegyptiaca with other anticancer drugs as the methotrexate to increase their efficacy and decrease their dose so their adverse effects could be decreased.
• Further studies should be done to clarify the role of B. aegyptiaca in prevention of liver fibrosis and fatty infiltration.