Evaluating Therapeutic Characteristics of Anticancer L - Glutaminase Enzyme on Ethylenimine Induced Mutagens in Rats
تقييم الخاصية الدوائية للأنزيم L-glutaminase المضادة للسرطان على طفرات المستحثة بمادة Ethylenimine في الجرذان

Gulbahar Karim, Ali H. Taqi
2018 Kirkuk University Journal-Scientific Studies  
This study include, the partial purification of L-glutaminase enzyme from E.coli by ammonium sulphate precipitation (40%-80%) fractions, and the enzyme activity elevated to 75 IU/ml following dialysis. Pharmacological evaluation of L-glutaminase enzyme was used for the in vivo studies on sixty rats as an animal models. The hematological parameters for rats in the treated group (T2) showed significant changes at the level (P<0.05) when compared with normal control group (T1). The WBCs was found
more » ... o be increased with the reduction in the RBCs and Hemoglobin (Hb.) concentration. However there were no significant changes of these parameters in the other treated groups (T3, T4, T5, and T6) in contrast to (T1). The serum total protein, globulin and, albumin were found to be decreased significantly for the (T2) in comparison with the (T1). However, their level had been relatively normalized in comparison to (T2) as a result of administrating doses of 0.5, and 1.0 ml L-glutaminase in combination with 0.5 mg ethylenimine /Kg. body weight for T5 and T6 respectively. Concerning the level of serum globulins, there were a significant elevation in the serum globulins value for T5 and T6 which were treated with doses of 0.5 and 1.0 ml L-glutaminase in combination with 0.5 mg ethylenimine/Kg. body weight respectively. While, a significant decrease in the values of serum albumin detected in all rats' treated groups and the L-glutaminase efficacy was not achieved completely compared with the T1.The Liver enzymes ALT, AST and, ALP for T2 showed significant elevation at the level (p<0.05) when compared with T1. The treatment with L-glutaminase at doses of 0.5 and, 1 ml /kg body weight in combination with 0.5 mg of ethylenimine/kg body weight in T5 and, T6 groups respectively, seem to reverse the changes to word near normal level. There were a Web Site: www.uokirkuk.edu.iq/kujss E. mail: kujss@uokirkuk.edu.iq 2 significant elevation (p<0.05) in the relative weight of Liver, kidney and spleen of rats in T2 in comparison with T1. However, there weight decreased significantly (p<0.05) in T5 group in comparison to (T2). Whereas there were no significant differences in the organs' relative weight for the T6 group from that of T1. The finding of the present study improved the preventive role of the L-glutaminase against the mutagenic, and carcinogenic effect of ethylinimine, and it was dose dependent. TWBCs ‫مع‬ ‫انخفاض‬ ‫الحمر‬ ‫الدم‬ ‫يات‬ ‫لكر‬ ‫الكمي‬ ‫العدد‬ ‫في‬ TRBCs ‫الهيموكموبي‬ ‫وتركيز‬ ‫ن‬ Hb concentration ‫لممجموعة‬ ‫المعاممة‬ T2 ‫لمادة‬ ‫الفموي‬ ‫يع‬ ‫التجر‬ ‫نتيجة‬ Ethylenimine ‫ة‬ ‫السيطر‬ ‫بمجموعة‬ ‫نة‬ ‫مقار‬ T1 ‫عمى‬ ‫معنويا"‬ ‫ا"‬ ‫تغير‬ ‫أ‬ ‫يطر‬ ‫لم‬ ‫حين‬ ‫في‬ ‫االخرى‬ ‫المجاميع‬ ‫في‬ ‫ات‬ ‫المتغير‬ ‫تمك‬ ، ‫معنوي‬ ‫انخفاض‬ ‫حصل‬ ‫كما‬ (P<0.05) ‫مصل‬ ‫في‬ ‫االلبومين‬ ‫و‬ ‫الكموبيولين‬ ‫و‬ ‫الكمي‬ ‫لمبروتين‬ Web Site: www.uokirkuk.edu.iq/kujss E. mail: kujss@uokirkuk.edu.iq 6 hemoglobin (Hb.) concentration, the manual Sahli's (Acid hematin) method has been used. Total Red Blood Cells (TRBC) count has been measured by standard manual technique. For measuring the Total White blood cells(TWBC) count the direct method has been used [16]. Biochemical parameters: The blood samples without EDTA were placed at room temperature for about one hour at room temperature, then centrifuged at 3000 rpm for 15 minutes. Follow that the supernatant (serum) was collected by using micropipette and transferred to ependorf tubes, then kept in a freezer at -20 °C for one week for biochemical tests. serum separator tubes were used for serum collection. Serum concentration of total protein, albumin, and globulin were determined according to Kits from Biolabo (France). The Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and Alkaline phosphatase (ALP) were determined according to the manufacturers recommended procedure from Biomerieux-France. The ALT test applied immediately following sample collection. ALT estimated immediately. Statistical Analysis: Data were analyzed by the ANOVA analysis, using the general linear model of the Statically Analysis System. Significant treatment differences were evaluated using Duncan's multiplerange test. All statements of significance are based on the 0.05 level of probability. The Results and Discussion:  Purification of L-glutaminase The Partial purification of the enzyme using ammonium sulphate precipitation showed that the best fraction was (80%) in contrast to the crude enzyme, 40% and 50% fractions. It produced the maximum value of enzyme activity that was 75 IU/ml following dialysis which estimated by Nesslerization process [14] . The finding of the present study was in agreement with that of other investigators who found that 80% ammonium sulphate was produced the highest yield of Lglutaminase activity from Streptomyces avermitilis [17]. Pharmacological Evaluation of L-glutaminase Enzyme: The partial purified L-glutaminase produced by E.coli isolate obtained from wound sample was used for in vivo study in rat models to investigate the effect of L-glutaminase enzyme on some hematological, chemical parameters and the relative organs weight of treated animal. Web Site: www.uokirkuk.edu.iq/kujss E. mail: kujss@uokirkuk.edu.iq 7 ALT, AST and ALP when compared to the treated group (T2) which administered ethylenimine (p<0.05), this finding may be attributed to the effect of L-glutaminase which may stabilize hepatocyte cell membrane and prevent delivery of ALT, AST, and ALP to the extracellular fluid. Since the elevated values of ALT and, AST in serum is indicative of liver damage [24] . ALT is elevated in acute liver damage and, is a more reliable marker of liver integrity than AST [25] . Since AST enzyme is not specific for liver, it is also present in RBCs, and cardiac with skeletal
doi:10.32894/kujss.2018.13.4.1 fatcat:wum3nigw3nfazcfqfnknvr2yuq