Jagetia Abhinav, Jagetia Ganesh Chandra, Jha Shalini
Department of Metallurgy, Malviya National Institute of Technology, Jaipur-302 017, India.
J Appl Toxicol. 2007 Mar-Apr;27(2):122-32. doi: 10.1002/jat.1175.
The effect of naringin, a grapefruit flavonone was studied on bleomycin-induced genomic damage and alteration in the survival of cultured V79 cells. Exposure of V79 cells to bleomycin induced a concentration dependent elevation in the frequency of binucleate cells bearing micronuclei (MNBNC) and a maximum number of MNBNCs were observed in the cells treated with 50 microg ml(-1) bleomycin, the highest concentration evaluated. This genotoxic effect of bleomycin was reflected in the cell survival, where a concentration dependent decline was observed in the cells treated with different concentrations of bleomycin. Treatment of cells with 1 mm naringin before exposure to different concentrations of bleomycin arrested the bleomycin-induced decline in the cell survival accompanied by a significant reduction in the frequency of micronuclei when compared with bleomycin treatment alone. The cell survival and micronuclei induction were found to be inversely correlated. The repair kinetics of DNA damage induced by bleomycin was evaluated by exposing the cells to 10 microg ml(-1) bleomycin using single cell gel electrophoresis. Treatment of V79 cells with bleomycin resulted in a continuous increase in DNA damage up to 6 h post-bleomycin treatment as evident by migration of more DNA into the tails (% tail DNA) of the comets and a subsequent increase in olive tail moment (OTM), an index of DNA damage. Treatment of V79 cells with 1 mm naringin reduced bleomycin-induced DNA damage and accelerated DNA repair as indicated by a reduction in % tail DNA and OTM with increasing assessment time. A maximum reduction in the DNA damage was observed at 6 h post-bleomycin treatment, where it was 5 times lower than bleomycin alone. Our study, which was conducted on the basis of antioxidant, free radical scavenging and metal chelating properties of naringin demonstrates that naringin reduced the genotoxic effects of bleomycin and consequently increased the cell survival and therefore may act as a chemoprotective agent in clinical situations.
研究了葡萄柚黄酮柚皮苷对博来霉素诱导的基因组损伤及培养的V79细胞存活率改变的影响。将V79细胞暴露于博来霉素中,会导致含微核双核细胞(MNBNC)频率呈浓度依赖性升高,在用50μg/ml(-1)博来霉素(评估的最高浓度)处理的细胞中观察到MNBNC的最大数量。博来霉素的这种遗传毒性作用反映在细胞存活率上,在用不同浓度博来霉素处理的细胞中观察到浓度依赖性下降。在暴露于不同浓度博来霉素之前,用1mM柚皮苷处理细胞可阻止博来霉素诱导的细胞存活率下降,与单独使用博来霉素处理相比,微核频率显著降低。发现细胞存活率与微核诱导呈负相关。通过单细胞凝胶电泳将细胞暴露于10μg/ml(-1)博来霉素来评估博来霉素诱导的DNA损伤的修复动力学。用博来霉素处理V79细胞导致博来霉素处理后6小时内DNA损伤持续增加,彗星尾部迁移更多DNA(%尾DNA)以及随后橄榄尾矩(OTM,DNA损伤指数)增加表明了这一点。用1mM柚皮苷处理V79细胞可减少博来霉素诱导的DNA损伤并加速DNA修复,随着评估时间的增加,%尾DNA和OTM降低表明了这一点。在博来霉素处理后6小时观察到DNA损伤最大程度降低,此时比单独使用博来霉素低5倍。我们基于柚皮苷的抗氧化、自由基清除和金属螯合特性进行的研究表明,柚皮苷降低了博来霉素的遗传毒性作用,从而提高了细胞存活率,因此在临床情况下可能作为一种化学保护剂。