الفهرس 
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Abstract
EFFECT OF PLANT PROTEASE INHIBITOR ON RATS TREATED WITH A CANCER INDUCER
VII-SUMMARY
Objectives.
The goals of this study were to investigate the effects of Bowman-Birk protease inhibitor (BBI) on male albino rats treated with AFB1 followed by CCl4 as cancer inducer.
Methods.
Four groups of 30 rats each were used:
Group I : (The control) injected with 0.9 NaCl solution, pH 7.2 or corn oil in case of using AFB1 or CCl4 for treating rats respectively.
Group II : (AFB1-CCl4 treatment) the rats were treated with AFB1 0.5 mg/kg B.W. each third day for 3 weeks and one week later rats were injected with CCl4 dissolved in corn oil (0.8ml/k.g B.W.) once a week for 8 weeks and left without treatment for another 4 weeks.
Group III : (AFB1-CCl4 followed by treatment with BBI) the animals were treated with AFB1 and CCl4 as described in Group II, during the last 4 weeks the rats were injected with BBI (40mg/K.g B.W.) each third day.
Group IV : (treated with BBI followed by AFB1-CCl4) received BBI during the first 4 weeks as described in Group III. Then treated with AFB1 and CCl4 for 12 weeks as described in Group II.
After 16 weeks of treatments, some biochemical measurements in serum and liver homogenate, some hematological, immunological parameters, and histopathological examinations of liver sections of different groups were performed. The Caspase-3 enzyme activity, apoptotic marker, as well as levels of GSH and GST were determined in liver tissues.
Results.
Effect on relative weight of some organs.
The relative weights (percent of body weight) of liver, kidney and spleen increased during the first 12 weeks in rats of Group II and III compared to the control group, but treatment with BBI before AFB1-CCl4 injection depressed the increases of relative weight in the above mentioned organs. On the other hand rats treated with AFB1-CCl4 (Group II, III and IV) demonstrated decreases in the relative weights of testes compared to the animals of the control group.
Effect on liver function tests.
During the first 12 weeks of the experiment the levels of transaminases (ALT and AST) were significantly higher in the two of the treated groups (Group II and III), but tend to decline as the work progressed. Animals of Group IV showed similar pattern to those of the control group.
Serum total bilirubin of animals treated with AFB1-CCl4 (Group II and III) was significantly higher than the control animals. Treatment with BBI before or after AFB1-CCl4 revealed very close values of total bilirubin compared to those of the control group.
Effect on kidney function tests.
During the first 12 weeks, the levels of urea increased from about 40 to 110 mg/dl in rats of Group II and III compared to 39 mg/dl in rats of the control group. Animals treated with BBI before subjecting to AFB1-CCl4 (Group IV) depressed the increase in urea levels. Creatinine level in serum of animals of the 4 groups demonstrated similar changes to those recorded for urea.
Effect on Caspase-3, GSH and GST in rat livers.
Livers of rats of Group II and III demonstrated higher levels of Caspase-3 activity during the first 12 weeks of the experiment, compared to the control animals, but the enzyme activity tend to decrease during the last 4 weeks. No significant difference in Caspase-3 levels in livers of rats of Group IV and those of the control animals.
Our results indicated increase in GSH levels in animals of Group II and III in comparison with the control group. But the values of GSH recorded in animals of Group IV were close to those observed for the control group.
The activity of GST in livers of rats treated with AFB1-CCl4 (Group II) decreased from 3.9 to 2.0 micro mol / mg protein / min by the end of the experimental work. Treatment with BBI before or after AFB1-CCl4 treatment increased the GST activity from 3.9 to 5.5 and from 3.9 to 7.2 micro mol / mg protein / min in livers of rats of Group III and IV respectively.
Effect on hematological parameters.
The results of hematological examinations indicated a depression in total red blood cells (RBCs), including hemoglobin content of the blood.
Effect on some immunological parameters.
1-Total leucocytes count:
Data revealed small increases in total leucocyte count during the first 4 weeks, and continued to raise at higher rates as time advanced in rats of groups treated with AFB1-CCl4 as well as BBI (Group III and IV). Similar increase in leucocyte counts was observed in Group II, which continued during the first 12 weeks but tend to decline by the end experimental period.
2- Differential count of leucocytes.
A- Lymphocytes and Neutrophils:
The results of deferential count of leucocytes of rats of different groups revealed increases in the percentage of lymphocytes and a concurrent decreases in the percentage of neutrophil in all treated groups in comparison to the control (Group I).
B- Monocytes:
Our results of monocytes count indicated that the percentage of monocytes ranged from 3 to 5 % of total leucocytes. The monocytes percentages increased in animals of Group II and III and to less extent in rats of Group IV compared to the control.
C-Eosinophil and Basophil:
One type of granulocytes, the neutrophile, is also a phagocyte; it uses its packaged chemicals to break down the microbes it ingests.
Eosinophils and basophils are granulocytes that ‘degranuloate’ spraying their chemicals onto harmful cells or microbes nearby.
Results of the present work indicated that the percentages of eosinophil and basophil were increased in all treated groups (Group II, III and IV) compared to the animals of the control group (Group I). Their percentages account for 2-4 % and 0.2-0.6 % of total leucocytes for eosinophil and basophil respectively.
3-Phagocytosis.
Compared to untreated group (control group), phagocytosis percentages in rats decreased during the first two weeks and then tend to increase as the time progressed to reach their maximum levels by the end of week 12 and week 14 for Group II and III respectively. The animals of Group IV revealed increase in phagocytosis through out the whole experimental period.
4- Chemokinesis.
Compared to the animals of the control group (Group I), samples of rats of Group II and III showed large increases in the values of mean chemokinitic indices up to week 12 but tend to decrease as the time advanced. Treatment with BBI before using the cancer inducer (AFB1-CCl4) seems to depress the increase in chemokinetic values.
5-Chemotaxsis and cell migration:
Different migration patterns were observed for the blood of rats of different treated groups (Group II, III and IV), but no chemotactic migration was recorded in the blood of Group I rats.
6-Macrophage migratin inhibition factor:
Results indicated that the peritoneal exudate cells from animals treated with AFB1- CCl4 (Group II) demonstrated inhibition of migration ranged from 51 to 73 % but the same cells from rats of control groups showed no inhibition. The injection with BBI before or after treatment with AFB1- CCl4 decreased changes in migration inhibition.
7-Total immunoglobulin (Ig):
The levels of total Ig in Group II and III were higher than those recorded for the control group (Group I). Injection with BBI before treatment with AFB1- CCl4 (Group IV) depressed the increase in total Ig.
CONCLUSION
AFB1-CCl4 treatment led to damage liver and kidney and may affect other organs (spleen and testes). AFB1- CCl4 affect the components of the immune system and cause apoptotic and necrobiotic changes in the liver. BBI exhibited and efficient protective effect against AFB1 and CCl4. Thus using BBI as chemopreventive agent against AFB1-CCl4 proved to be effective. But more work needs to be done before using for human.