الفهرس 
Introduction and Aim of the WorkOnly 14 pages are availabe for public view
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Abstract
Cancer is the second largest cause of death after cardiovascular diseases in the world, and therefore many preclinical and clinical scientific studies have been conducted to find safe treatments for cancer patients. Chemotherapy is one of the most important current treatments, but it has significant harmful effects on various tissues throughout the body, especially in both the kidneys and liver. Therefore, it was necessary to work on finding other treatments derived from natural products from plant and animal sources that have anti-cancer ability and do not have a side effect on the tissues and organs of the body. Therefore, this study aimed to evaluate the effect of scorpion extract (LQWBE) as an antitumor agent in vitro using both hepatic tumor cells (HepG-2) and human mammary tumor cells (MCF-7), and also to study the antitumor effect of LQWBE extract on mice. Carriers of abdominal ascites tumor, in addition to its protective effect on the livers of mice, as a histological study using H&E stain, as well as IHC study using caspase-3 markers, and were examined by light microscopy. The percentage of protein, fat and total carbohydrates of LQWBE were also measured and separated by electrophoresis. One hundred and twenty female mice with a weight of approximately 25 ± 3 gm were used. These mice were divided into two main experiments, with 60 mice in each experiment. Each experiment was divided into four groups of equal number, each separately, and each group contained fifteen (15) mice as follows: The first experiment: This experiment was done to study the effect of scorpion extract (LQWBE) as a protection against mice liver changes. The first group: (control group), the mice were injected with saline solution in the abdominal cavity (200 μL/mouse) for 6 consecutive days. The second group: (cisplatin group only (Cis), where the mice were injected at a dose of 2 mg/kg/mouse for 6 consecutive days. The third group: (scorpion extract group (LQWBE), where the rats were injected with scorpion extract at a dose of 2 mg/kg/rat for 6 consecutive days. The fourth group: the group treated with cisplatin and scorpion extract together (Cis/LQWBE) at the same previous doses for 6 consecutive days. The mice of the first experiment were dissected on the fourteenth day from the beginning of the experiment. The second experiment: This experiment was performed in order to study the anti-tumor effect of scorpion extract (LQWBE) on mice bearing EAC-induced ascites tumor. The mice were injected with 1X106 EAC tumor cells, and they were divided into four groups each group included fifteen (15) mice as follows: The first group: (EAC group), the mice were injected with tumor EAC, where each mouse was injected with 200 μL of the saline solution daily for 6 consecutive days. The second group: (EAC/Cis group) which represented the EAC- bearing mice tumor group treated with cisplatin (Cis) at a dose of 2 mg/kg/mouse for 6 consecutive days. The third group: (EAC / LQWBE group) which represented EAC- bearing mice tumor group injected injected with LQWBE at a dose of 100 mg/kg/mouse for 6 consecutive days. The fourth group: (EAC/Cis / LQWBE group), EAC- bearing mice tumor group injected with cisplatin and scorpion extract (LQWBE) together at the same doses mentioned before for 6 consecutive days. The mice of the second experiment were dissected on the fourteenth day from the beginning of the experiment. Blood samples were collected at the end of the two experiments to make a complete blood picture and measure liver enzymes and kidney functions. Also, samples were taken from the liver tissues of all mice groups to examine histological and IHC changes. The results are summarized as follows: - 1- Analysis of Scorpion Extract LQWBE: The present results showed that the percentage of total protein, fat, and carbohydrates were 12.5, 20, and 30 mg/g, respectively. The analysis of the protein pattern of LQWBE revealed the presence of 6 bands that ranged between 15-70 kilo Daltons. The results showed that the potassium concentration was 9.1 ± 0.16 mg/g and the sodium and calcium concentrations were 9.1 ± 0.16 and 5.5 ± 0.77 mg/g, respectively. While the concentrations of Mg were 0.35 ± 0.28, and Zn 0.02 ± 0.02 mg/g respectively, gas chromatography-mass spectrometry (GC-MS) analysis also showed that oleic acid had a maximum peak area (14.95%) at a retention time (RT) of 22.98 minute. 9,12-octadecanoic acid - ethyl ester, 9-octadecanoic acid, methyl ester, linoleic acid, and hexadecanoic acid, methyl ester were detected at RTs 22.21, 23.32, 22.30, 22.90 and 19.54 min, with peak areas (% ) 14.59, 9.39, 9.98, 7.66 and 6.24%, respectively. 2- Results of determining the lethal half concentration of the extract on tumor cell lines in vitro: The results also showed the effect of the dose that killed half of the hepatic tumor cells (HepG-2) was 992 and greater than 1000 μg/mL. 3- Effect results of EAC / Cis extract: The results showed a decrease in the weight of the mice that were injected with Cis when compared to the control group. • After treatment with LQWBE, an increase in weight was observed compared to the control group. • As for the groups that were injected with tumor cells (EAC), they showed a significant increase in body weight • Treating tumor-bearing (EAC) mice with Cis showed a significant decrease in body weight. • LQWBE showed an improvement in body weight. • As for the EAC tumor-bearing mice, which were treated with both Cis/ LQWBE, their weights returned to the normal range. • The results also showed a significant decrease in the total volume of the tumor and the number of tumor cells in tumor-bearing (EAC) mice that were treated with Cis drug compared to tumor-bearing (EAC) mice. • The results also showed that the use of LQWBE has led to a significant decrease in the number of tumor cells, which indicates its anti-tumor activity compared to tumor-bearing (EAC) mice. • The tumor-bearing (EAC) mice that were treated with both Cis/LQWBE together showed a significant decrease in the number of tumor cells compared to the (EAC) tumor-bearing mice. 4- Hematology results: • The treatment of normal normal mice with Cis or LQWBE or both Cis/ LQWBE did not affect the number of red blood cells (R.B. Cs), hemoglobin (Hb) and hematocrit (Hct) when compared to the control group. The results also showed a decrease in the number of white blood cells (WBCs) in the group of mice that were injected with Cis, compared to the control group. • The percentages of the mice group injected with Cis had an increase in neutrophils and a decrease in monocytes and lymphocytes. • The treatment of mice with LQWBE has led to the return of those previous values to their normal levels. Similar results were also recorded in the group of mice injected with Cis drug. It indicated an increase in the number of platelets and that treatment with LQWBE or Cis/LQWBE led to a return to normal values. • There was no significant change in the number of RBCs, hematocrit (Hct), hemoglobin (Hb) and platelets in tumor-bearing mice treated with Cis or LQWBE or both. 5- Biochemical results: • The results recorded an increase in the levels of aminotransferase enzymes, such as ALT and AST of the mice group injected with Cis when compared to the control group. While the mice group treated with LQWBE or Cis /LQWBE together recorded a decrease in aminotransferase levels. • The results also demonstrated an increase in the levels of urea and creatinine for the group of rats injected with Cis drug compared to the control group. While the mice group treated with LQWBE or Cis /LQWBE together showed a decrease in their levels which proves an improvement in kidney function. • As for the tumor-bearing mice (AEC), the results showed a significant increase in the levels of ALT and AST enzymes, but after treatment with Cis, the results showed a significant decrease in the activity of these enzymes, and the treatment with LQWBE showed a significant decrease in these two enzymes, while the mice treated with LQWBE/Cis together showed a decrease in the measurements to almost normal. • The results showed that tumor-bearing mice (AEC) increased significantly in urea and creatinine, while in the group of mice that were injected with LQWBE or Cis, a significant decrease was observed in urea and creatinine, while treatment with LQWBE / Cis together led to a decline in the measurements to almost normal. 6- Histological observations of the liver: The histological examination of the livers of the control mice group light microscopy showed that normal architecture of the liver cells with homogeneous cytoplasm and healthy round nuclei, and the appearance of the normal central and portal veins was normal as well as the Kupffer cells located in the walls of the blood sinusoids are normal. The livers of mice injected with cisplatin (Cis) showed many changes in the structure of the liver tissues as a vacuolation of the cytoplasm, karyolitic nuclei, appearance of necrosis and death of some hepatocytes as well as the appearance of dilatation of the blood vessels, central and portal veins, and an increase in the activity of Kupffer cells. On the other hand, the treatment with LQWBE alone or LQWBE/Cis together showed significant improvement, recovery of the normal form of most hepatocytes with binucleated cells, central and portal and conclusion - 119 - 2- Results of determining the lethal half concentration of the extract on tumor cell lines in vitro: The results also showed the effect of the dose that killed half of the hepatic tumor cells (HepG-2) was 992 and greater than 1000 μg/mL. 3- Effect results of EAC / Cis extract: The results showed a decrease in the weight of the mice that were injected with Cis when compared to the control group. • After treatment with LQWBE, an increase in weight was observed compared to the control group. • As for the groups that were injected with tumor cells (EAC), they showed a significant increase in body weight • Treating tumor-bearing (EAC) mice with Cis showed a significant decrease in body weight. • LQWBE showed an improvement in body weight. • As for the EAC tumor-bearing mice, which were treated with both Cis/ LQWBE, their weights returned to the normal range. • The results also showed a significant decrease in the total volume of the tumor and the number of tumor cells in tumor-bearing (EAC) mice that were treated with Cis drug compared to tumor-bearing (EAC) mice. • The results also showed that the use of LQWBE has led to a significant decrease in the number of tumor cells, which indicates its anti-tumor activity compared to tumor-bearing (EAC) mice. • The tumor-bearing (EAC) mice that were treated with both Cis/LQWBE together showed a significant decrease in the number of tumor cells compared to the (EAC) tumor-bearing mice. Hematology results: • The treatment of normal normal mice with Cis or LQWBE or both Cis/ LQWBE did not affect the number of red blood cells (R.B. Cs), hemoglobin (Hb) and hematocrit (Hct) when compared to the control group. The results also showed a decrease in the number of white blood cells (WBCs) in the group of mice that were injected with Cis, compared to the control group. • The percentages of the mice group injected with Cis had an increase in neutrophils and a decrease in monocytes and lymphocytes. • The treatment of mice with LQWBE has led to the return of those previous values to their normal levels. Similar results were also recorded in the group of mice injected with Cis drug. It indicated an increase in the number of platelets and that treatment with LQWBE or Cis/LQWBE led to a return to normal values. • There was no significant change in the number of RBCs, hematocrit (Hct), hemoglobin (Hb) and platelets in tumor-bearing mice treated with Cis or LQWBE or both. 5- Biochemical results: • The results recorded an increase in the levels of aminotransferase enzymes, such as ALT and AST of the mice group injected with Cis when compared to the control group. While the mice group treated with LQWBE or Cis /LQWBE together recorded a decrease in aminotransferase levels. • The results also demonstrated an increase in the levels of urea and creatinine for the group of rats injected with Cis drug compared to the control group. While the mice group treated with LQWBE or Cis /LQWBE together showed a decrease in their levels which proves an improvement in kidney function. • As for the tumor-bearing mice (AEC), the results showed a significant increase in the levels of ALT and AST enzymes, but after treatment with Cis, the results showed a significant decrease in the activity of these enzymes, and the treatment with LQWBE showed a significant decrease in these two enzymes, while the mice treated with LQWBE/Cis together showed a decrease in the measurements to almost normal. • The results showed that tumor-bearing mice (AEC) increased significantly in urea and creatinine, while in the group of mice that were injected with LQWBE or Cis, a significant decrease was observed in urea and creatinine, while treatment with LQWBE / Cis together led to a decline in the measurements to almost normal. 6- Histological observations of the liver: The histological examination of the livers of the control mice group light microscopy showed that normal architecture of the liver cells with homogeneous cytoplasm and healthy round nuclei, and the appearance of the normal central and portal veins was normal as well as the Kupffer cells located in the walls of the blood sinusoids are normal. The livers of mice injected with cisplatin (Cis) showed many changes in the structure of the liver tissues as a vacuolation of the cytoplasm, karyolitic nuclei, appearance of necrosis and death of some hepatocytes as well as the appearance of dilatation of the blood vessels, central and portal veins, and an increase in the activity of Kupffer cells. On the other hand, the treatment with LQWBE alone or LQWBE/Cis together showed significant improvement, recovery of the normal form of most hepatocytes with binucleated cells, central and portal venous dilatation regressed to normal, and most of the Kupffer cells returned to their usual state with a few in an active state. As for the group of mice carrying tumors (EAC), the results showed significant changes in the structure of the hepatic tissues, necrosis and death of most liver cells, the presence of vacuolation in the cytoplasm, disintegrating nuclei, a large crowding of infiltration of the inflammatory cells around the congested and dilated central and portal vein, widening of the bile ducts, and hypertrophy of the Kupffer cells. On the other hand, treatment with either Cis or LQWBE alone or Cis/LQWBE together showed a significant improvement in the liver tissues, the changes decreased, and the hepatocytes recovery their usual status appeared almost normal in rows from the central vein with intact nuclei especially in the mice treated with the Cis/LQWBE together. 7- IHC results: The IHC results of the livers of the normal control mice group by using caspase-3 markers and light microscopy expressed a normal, weak or rare immunostain to hepatocytes. The liver sections the mice group injected with cisplatin (Cis) expressed strong immunostain in most of the hepatic cytoplasm while the mice group treated with LQWBE only expressed few immunostain to apoptotic hepatocytes. The mice treated with both Cis /LQWBE expressed a noticeable deficiency f and a reduction of immunopositive stain to hepatocytes, and few apoptotic cells are still present. The IHC results of the livers of tumor-bearing mice (EAC) showed a positive, highly strong stained, dark image of the cytoplasm of most hepatocytes using caspase-3 markers due to the abundance of programmed hepatocytes death, while the mice treated with EAC/Cis or EAC/LQWBE showed a decline in the immunostain to hepatocytes. The treatment of tumor-bearing mice (EAC) group with both Cis/LQWBE together showed a clear improvement and a significant decrease in the immunostain of the hepatocytes to caspase-3 markers, and the restoration of the appearance of the liver tissues similar to the normal ones with rare apoptotic hepatocytes.. Conclusion and recommendation: The current study concluded that the protein content of LQWBE is higher than the fat content, and there is no carbohydrate content. Moreover, the present study recorded that there are six bands of protein using the method of protein electrophoresis. where by using the tumor abdominal ascites model of mice, it was found that the extract of LQWBE has antitumor activity, as well as a protective effect against toxicity induced by cisplatin, and it also can improve liver tissue and decreases the apoptosis by decreasing the rate of caspase-3. Therefore, the results recommend that more studies must be done on LQWBE extract, as well as the separation of biologically active compounds from it, to test them as antitumors in the laboratory and on experimental mouse models.