Assessment of the effect of cell transplantation on DNA repair in rat hepatocytes exposed to carbon tetrachloride (CCl4) by DNA comet assay

L. V. Shabasheva, G. Protasova, V. B. Popov
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Abstract

Introduction. The effect of carbon tetrachloride (freon-10, asordin, hladon-10) is an organochlorine compound with the chemical formula CCl4 and the subsequent transplantation of fetal liver cells (FLC) on DNA degradation and repair in rat hepatocytes by means of alkaline single-cell gel electrophoresis (DNA comet assay) was assessed. Material and methods. Acute toxic damage to the rat liver was simulated by a single oral administration to female Wistar rats of CCl4 in an oil solution at a dose of 3000 mg/kg. As a protective agent, a suspension of FLC of E19 rat fetuses was used. Quantitative assessment of the degree of damage to the nuclear DNA of liver cells was performed by DNA comet assay on days 1, 3, 5, 7 and 16 of the experiment. Results. Intravenous injections of fetal liver cells 6 h after exposure to CCl4 induces DNA repair processes in rat hepatocytes in 5-7 days and led to a decrease in the intensity of nuclear DNA damage. The trend toward a decrease in the number of undamaged hepatocytes continued on the 16th day of the experiment, and, therewith, the enhancement of reparative processes after FLC injection revealed itself in in a significant decrease in the number of hepatocytes with a high intensity of nuclear DNA damage. Limitations. To prevent unwanted death of animals in the group, studies were limited to a dose of 3000 mg/kg of CCl4 in oil solution. Conclusion. The method of alkaline single-cell gel electrophoresis (DNA comet assay) allowed quantitative assessment of the degrees of genome damage and repair. The obtained positive results suggest that FLC exert a protective effect of the structure of the DNA of rat liver role after acute exposure to CCl4.
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细胞移植对四氯化碳(CCl4)暴露大鼠肝细胞DNA修复的影响
介绍。采用碱性单细胞凝胶电泳(DNA彗星法)研究了化学式为CCl4的有机氯化合物四氯化碳(氟利昂-10、蛇毒素、hladon-10)和胎儿肝细胞移植对大鼠肝细胞DNA降解和修复的影响。材料和方法。雌性Wistar大鼠单次口服剂量为3000 mg/kg的CCl4油溶液,模拟大鼠肝脏急性毒性损伤。采用E19大鼠胎儿FLC悬浮液作为保护剂。在实验第1、3、5、7、16天采用DNA彗星法定量评价肝细胞核DNA的损伤程度。结果。暴露于CCl4后6小时静脉注射胎儿肝细胞可诱导大鼠肝细胞在5-7天内的DNA修复过程,并导致核DNA损伤强度降低。实验第16天,未损伤的肝细胞数量继续减少,因此,FLC注射后修复过程的增强表现为核DNA高强度损伤的肝细胞数量显著减少。的局限性。为了防止该组动物意外死亡,研究仅限于在油溶液中使用3000 mg/kg的CCl4剂量。结论。碱性单细胞凝胶电泳(DNA彗星测定)方法可以定量评估基因组损伤和修复的程度。结果表明,FLC对大鼠急性CCl4暴露后肝脏DNA结构具有保护作用。
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