الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
The marine environment is a promising area for exploring biologically active natural products, which are a rich source of new treatments. Biologists believe that ascidia produces a rich mix of biologically active compounds from the tunic and it’s biologically active compounds contain important biological agents, including anti-inflammatory, antibacterial, and viral properties with toxic properties of pathogenic cells, anti-cancer properties, the first anticancer product was Trabectedin (ET-743 Yondeli), which is a product of Ecteinascidia turbinate, found along the Suez Canal in Egyptian beaches. Ascidia (same-sac family) is a family of tunicates, that animals feeds on filtration can expel water when it is threatened, it’s oval body, covered with a solid cellulose mantle, with many round warts, folded grooves, It is a link between vertebrates and invertebrates, represented in spinal cord in the swimming larva (characteristic of vertebrates), which quickly disappears when the larva settle in appropriate spot. One member of this family is Styela Plicata, which has not been mentioned in many Egyptian researches as biological activities, so this research will focus on it. The samples were collected from two Egyptian regions, Alexandria from the Eastern Port, and Ismailia from Lake Tamasah, which were kept frozen in the laboratory for further testing, for Summary 127 Extraction preparation ascidia were cut into small pieces, drying, grinding, to make powdered. According to the scientific apparatus used, an extract of ethanol was used for the GC-MS, water extract was used High pressure liquid chromatography (HPLC). Water extract for Egyptian Styela plicata have been chosen for non-toxicity properties of water but researches did not talk about IC50 for the crude Egyptian Styela plicata extract, so toxicity test was extremely needed for in vivo experiment. Experimental protocols: The study was divided into two experiments: Experiment (1) Toxicity test was terminated after 4 weeks, and Experiment (2) after 40 weeks. Experiment (1) In the Toxicity test, rats divided as follows: a) group 1 (5 rats) were administered oraly (1ml/day) with (2600 mg/kg) high dose of Styla plicata. b) group 2 (5 rats) were injected with (1300 mg/kg) diluted dose. c) group 3 (5 rats) were injected with (650 mg/kg). d) group 4 (5 rats) were injected with (325 mg/kg). e) group 5 (5 rats) were injected with (162.5 mg/kg). f) group 6 (5 rats) were receiving an equivalent volume of distilled water (negative control). Summary 128 Experiment (2) According to the results from experiment (1), which have shown the safety of the high-saturated dose. The saturated dose have been chosen for the second experiment to see if it had effect on colorectal cancer: a) group 1 (10 rats) were non-treatment normal control group (injected with 0.09% saline). b) group 2 (20 rats) were injected with DMH (50 mg/kg b.wt., sc., once a week for 8 weeks) and served as positive control. c) group 3 (20 rats) were injected were injected with DMH then administered oraly with fresh Styela water extract. d) group 4 (10 rats) were oraly administered with Styela extract, (1ml/day) daily for 40 weeks. Examinations were performed using the following methods: • Histopathology, tumuor biomarker (ACF) and tumour evaluation. • Immunohistochemistry PCNA. • RT-PCR: mRNA extraction, formation, SOD and NFkB genes expression. • blood biochemistry. The result obtained could be summarized as follows: Blood biochemistry: Experiment one Generally no significant differences were recorded with haemoglobin (HGB); mean corpuscular haemoglobin (MCH); content of mixed cells (monocyte, basophiles, eosinophils and neutrophils (NEUT) in treated groups in relative to control ones (received distal water) (P > 0.05). On the other side, the counts of white blood cells and lymphocyte (LYM) were significantly increased only at solely doses (162.5 mg/kg), red blood cells significantly decreased at 650 mg/kg dose, platelets count (PLT) significantly increase at 325 mg/kg, while decreased at 650 mg/kg dose. The entire previous significant were individually and there were no any significant in high doses indicating normal haemopoiesis and was not toxic to circulating red cells or platelets nor interfered with their production. The mean corpuscular volume (MCV) is an index of the size of the RBCs, when it is below normal; the RBCs will be smaller than normal and are described as microcytic. In examined rats the previous parameter was decreased than control ones but still within the normal index especially the concentration of the haemoglbin in corpuscle (MCHC) didn’t decrease but increased from G2-G5 indicated non deficiency of haemoglbin concentration recorded no detectable alterations in the haematological parameters of male western albino rats administered the ethanolic extract (up to 2000mg/kg bwt) of Ascidian. Summary 130 The results of liver function parameters like direct bilirubin (DB), aspirate amino transferase (AST) and albumin (ALB) were all within the normal limits and did not differ from control, Total bilirubin was increased with G1, G2 and G4 in comparison to control but still within the normal limits). Normal activities of amino transferases (AST) indicated non hepatotoxicity and lower amount of alanine transaminase (ALT) and alkaline phosphatase (ALK) confirmed the non toxicity of S. plicata extract. The normal values of the renal Biochemical parameters (urea, sodium, potassium and calcium) suggested that the extract didn’t produce any sort of disturbance in the renal function, It was significant increased in G1 and G3 (2600 and 650mg/kg body wt), but this elevation within the normal limit, ). Triglycerides also improved, they were significantly increased in higher doses than normal. Experiment two Blood analysis, there were no significant between groups, except in g2 injected with DMH was significantly decrease in Red blood cells, Platelets, Lymphocytes, where was significantly increase in Neutrophils compared to g1 control saline. Liver function, there were no significant different between groups, exept g2 DMH was significantly increase in ALT, and decrease in Total bilirubin, Total protein and Albumin. In Renal function, there were significantly decrease in HDL in g2 DMH compared with g1 control saline, but g3 (DMH+ASCex) Summary 131 was significantly decrease in UREA, cholesterol and LDL compared with G 2 DMH. PCNA labeling indices detected by immune histochemistry (IHC) showed significant inhibition in the groups treated with ASCex. Moreover, Also, expression of PCNA labeling indexes (%) were significantly higher in the DMH-treated epithelium of group (2), however treatment with ASCex significantly reduced the L1% in group (3). The L1% of group (4) treated with ASCex alone was almost similar to that of the control group (4). Moreover, after 40 weeks, the PCNA L1 (%) was significantly higher in the DMHtreated epithelium and in tumours than those treated with ASCex alone or after DMH administration. PCNA labeling indices (%) in experiment (1) were slightly higher in the DMH-treated colonic epithelia, albeit without statistical significance when compared with the other groups. The RT-PCR data for SOD and NFkB genes. The mRNA expression of SOD and NFkB was up-regulated after injection with DMH.