دراسة وراثية خلوية وجزيئية للتاثير الوقائي للكركم في سمية العقار ميترونيدازول في الفئران البيض == Cytogenetic And Molecular Study of Protective Effect of Curcumin In Toxicity of Metronidazole In White Mice
Author name:
فائق ابراهيم علي محمود
Supervisor name:
عادل فوزي شهاب
General topic:
Biology
Specific topic:
zoology - Genetics
Degree:
Doctorate
University:
Tikrit University - Faculty Of Education - Department Of Biology
Language:
Arabic
University location:
Salahaddin
First pages:
24T2933 - p.pdf
Abstract:
صممت الدراسة الحالية لغرض التعرف على التاثيرات السمية الوراثية والسمية الخلوية للعقار ميترونيدازول(MTZ) في الفئران البيض Balb/ C والتاثير الوقائي الذي توفره المعاملة المسبقة بالكركم(CUR) في سمية العقار ميترونيدازول. تم استخدام مؤشرات الوراثة الخلوية (النو | The current study was designed to identify the genotoxicity and cytotoxicity of Mitronidazole (MTZ) and protective effect of Curcumin (CUR) against the toxicity of MTZ in whit mice. Cytogenetic endpoints (micronuclei (MN) and mitotic chromosomes), Productive biomarkers (sperms), molecular cytogenetic biomarkers (comet assay), molecular endpoints (genomic and mitochondrial DNA concentration in liver and spleen and testis) and mutation assessment in mitochondrial DNA (mtDNA) of liver and testis by PCR and restriction pattern were used to evaluate genotoxicity, cytotoxicity of MTZ and protective effects of CUR in white Balb/Cmice. The study was conducted in the laboratories of college of education, college of science in Tikrit university and laboratories in the state company for drugs industry - Samarra - Iraq (SDI). Dosages of 30 and 50 mg/kb.b.wt. of MTZ were tested for genotoxic and cytotoxic and mutagenic effects in somatic and sperms of white mice. The dosages 4.5 and 10 mg/kgb.wt. of CUR were used to study the protective effects of CUR in genotoxicity and cytotoxicity and mutagenecity of MTZ. The current study showed that MTZ has genotoxic effect expressed by high frequency of micronuclei in polychromatic erythrocytes (PECs) which reached 15.5 ± 2.88 with the dosage 30 mg/kg.b.wt. and 21.31 ± 4.2 with the dosage 50mg/kg.b.wt. compared with 7.25 ± 1.88 in negative control. In the study of the protective effects of CUR against genotoxic and cytotoxic effects of MTZ, we observed that in mice treated with the dosage 4.5 mg/kg.b.wt of CUR for 9 days before the dosage 50 mg/kg.b.wt. of MTZ, There were significant p<0.05 decrease in MN frequency which reached 13.25 ± 4.19 compared with 21.31 ± 4.2 for mice treated with the dosage 50mg/kg.b.wt. of MTZ withoutCurcumin. The study showed that MTZ has cytotoxicity expressed by significant p<0.05 decrease in mitotic index (MI) which reached 0.010 in bone - marrow of the mice treated with 50 mg/kg.b.wt. compared with 0.023 in negative control. The dosage 4.5 mg/kg.b.wt. of CUR for 9 days before giving the dosage 50 mg/kg.b.wt. of MTZ resulted in returning of MI in bone - marrow 0.021 to its levels in negative control 0.023 compared with its value which reached 0.010 with the dosage 50 mg/kg.b.wt. of MTZ. The two dosages 30 and 50 mg/kg.b.wt. of MTZ caused significant p<0.05 decrease in total number of sperms which reached 2.4±2.4 and 1.5 ± 0.1 milions simultaneously compared with 6.7 ± 0.9 millions in negative control. More over there were significant p<0.05 decrease in motile sperms which reached 25.2 ± 11.0 and 14.6 ± 10.6 compared with 57.6 ± 8.0 of negative control. Significant p<0.01 decrease in the length of mid - piece and total length of sperm tail with the dosage 50 mg/kg.b.wt. of MTZ which reached 19.2 ± 6.0 and 69.0 ± 6.0 simultaneously compared with 31.6 ± 9.0 and 100.3 ± 15.0for negative control. Treating with the dosage 4.5 mg/kg.b.wt. of CUR for 9 day before the dosage 50 mg/kg.b.wt. of MTZ resulted in the return of mid - piece and total length of sperm tail which reached 29.5 ± 3.0 and 95.0 ± 5.0 close to the levels of negative control 31.06 ± 9.0 and 100.3 ± 15.0 simultaneously. Compared to 19.2 ± 6.0 for the length of mid - piece and 69.0 ± 6.0 for total length of sperm tail in mice treated with the dosage 50 mg/kg.b.wt. of MTZ without previous treatment with CUR. The study of damage levels in DNA with comet assay showed that MTZ induced significant p<0.01 levels of damage reached 45.4 ± 2.36 with the dosage 50mg/kg.b.wt. of MTZ in bone - marrow and 36.8 ± 0.58 in liver cells compared with 9.0 ± 2.12 and 5.2 ± 0.58 simultaneously in negative control. There were decrease occurred in the level of damage in DNA of bone - marrow and liver cells. However it did not return to its levels of negative control 9.0 ± 2.12 in bone - marrow and 5.2 ± 0.58 in liver cells. OTM values reached 26.4 ± 1.29 and 24.0 ± 1.14 in mice treated with the dosage 4.5 mg/kg.b.wt. of CUR for 9 days before the dosage 50 mg/kg.b.wt. of MTZ. However The values of OTM lower than its values in bone - marrow 45.4 ± 2.36 and 36.8 ± 1.66 in liver cells of mice treated with the dosage 50 mg/kg.b.wt. without CUR. In the molecular study, The results showed mutagenic effect of the dosage 50 mg/kg.b.wt. of MTZ in mice mtDNA expressed by novel restriction sites for Bam - HI and Hind - III in liver tissue and SauA3 in testis. The treatment of mice with the dosage 4.5mg/kg.b.wt. of CUR for 9 days before the dosage 50mg/kg.b.wt. of MTZ did not resulted in the inhibition of mutagenesis in mtDNA. In conclusion, results of this study showed that MTZ has genotoxic and cytotoxic effects in white mice and mutagenic effect in mice mtDNA. The dosage 4.5 mg/kg.b.wt.of Curcumin has protective effect against genotoxic and cytotoxic effects of MTZ. on the other hand the dosage 4.5 mg/kg.b.wt. of Curcumin has no protective effect against mutagenicity of MTZ in mice mtDNA.
