Usha Subbiah, Johnson Irudayam Maria, Malathi Raghunathan
Department of Genetics, Dr. ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai 600 113, India.
Mol Cell Biochem. 2006 Mar;284(1-2):57-64. doi: 10.1007/s11010-005-9013-6.
Time correlated Single Photon Counting study (TCSPC) was performed for the first time to evaluate the effect of resveratrol (RES) and genistein (GEN) at 10-100 microM and 10-150 microM respectively, in modulating the DNA conformation and the variation induced due to intercalation by the dyes, ethidium bromide (EtBr) and acridine orange (AO). It is demonstrated using UV-absorption and fluorescence spectroscopy that RES and GEN, at 50 microM and 100 microM respectively can bind to DNA resulting in significant de-intercalation of the dyes, preventing their further intercalation within DNA. Hyperchromicity with red/blue shifts in DNA when bound to dyes was reduced upon addition of RES and GEN. DNA-dependent fluorescence of EtBr and AO was quenched in the presence of RES by 87.97% and 79.13% respectively, while similar quenching effect was observed for these when interacted with GEN (85.52% and 83.85%). It is found from TCSPC analysis that the higher lifetime component or constituent of intercalated dyes (tau(2), A (2)) decreased with the subsequent increase in smaller component or constituent of free dye (tau(1), A (1)) after the interaction of drugs with the intercalated DNA. Thus these findings signify that RES and GEN can play an important role in modulating DNA intercalation, leading to the reduction in DNA-directed toxicity.
首次进行了时间相关单光子计数研究(TCSPC),以评估白藜芦醇(RES)和染料木黄酮(GEN)分别在10 - 100微摩尔和10 - 150微摩尔浓度下,对DNA构象的调节作用以及由溴化乙锭(EtBr)和吖啶橙(AO)插入所诱导的变化。通过紫外吸收光谱和荧光光谱证明,RES和GEN分别在50微摩尔和100微摩尔浓度时可与DNA结合,导致染料显著去插入,阻止其进一步插入DNA。加入RES和GEN后,与染料结合时DNA出现的增色效应及红/蓝移减小。在RES存在时,EtBr和AO依赖DNA的荧光分别猝灭87.97%和79.13%,而当它们与GEN相互作用时也观察到类似的猝灭效应(85.52%和83.85%)。从TCSPC分析发现,药物与插入DNA相互作用后,插入染料的较高寿命组分或成分(τ(2), A (2))随着游离染料较小组分或成分(τ(1), A (1))的随后增加而降低。因此,这些发现表明RES和GEN在调节DNA插入方面可发挥重要作用,从而导致DNA定向毒性降低。