大黄素蒽醌衍生物介导KB及KBv200细胞氧化损伤的研究

Oxidative damage of parental drug-sensitive KB cells and multidrug resistant KBv200 cells mediated by anthraquinone derivatives

目的 研究以大黄素为母体结构 ,人工半合成的 4种蒽醌衍生物抑制 KB和 KBv2 0 0细胞增殖的作用机制。方法 采用 MTT比色法测定细胞毒作用 ;采用流式细胞仪检测分别用 DCFH- DA和 Di OC6 荧光探针标记的细胞内活性氧 (ROS)和线粒体跨膜电位 (ΔΨm)。结果 4种蒽醌衍生物均能不同程度地抑制 KB和 KBv2 0 0细胞的增殖 ,表现为较强的体外抗肿瘤作用 :1,8-二 (二甲氨乙氨基 ) - 3-甲基 - 6 -甲氧基蒽醌 (H- 19)对 KB和 KBv2 0 0细胞 IC50 分别为 1.37和 1.4 2μmol/ L ;1-吡啶乙氨基 - 3-甲基 - 6 ,8-二甲氧基蒽醌 (H- 2 1)的 IC50 分别是 13.0和17.9μm ol/ L;1-吡咯烷乙氨基 - 3-甲基 - 6 ,8-二甲氧基蒽醌 (H- 2 5 )的 IC50 分别是 8.5和 11.7μmol/ L;1-羟丁氨基 -3-甲基 - 6 ,8-二甲氧基蒽醌 (H - 2 8)的 IC50 分别是 7.6和 8.6μmol/ L ,且各药对敏感株 KB和相应的多药耐药(MDR)细胞 KBv2 0 0的 IC50 相近 (P>0 .0 5 )。用 4种药物分别处理两种细胞 12、2 4、4 8h,12 h即能引起 ROS的明显增加 ,2 4 h达到最大 ,与 4 8h引起的 ROS增加差异不显著 ;各药均引起两种细胞ΔΨm时间依赖性的降低 ,4 8h时 ΔΨm降低达到最大。结论 4种大黄素蒽醌衍生物可能通过诱导细胞内 ROS的增加 ,使得 ΔΨm降低 ,从?

Object To study the inhibitory mechanism of four anthraquinone derivatives, which were obtained from modifying the structure of emodin, on proliferation of parental drug-sensitive KB cells and multidrug resistant (MDR) KBv200 cells. Methods Cytotoxicity was determined by tetrazolium (MTT) assay. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (ΔΨ m) in cells respectively labelled by DCFH-DA and DiOC6 were assayed by flow cytometry. Results All four anthraquinone derivatives suppressed proliferation of KB and KBv200 cells, showed potent cytotoxicity, the mean IC 50 of H-19 (1,8-dimethylaminethylamine-3-methyl-6-methoxy-anthraquinone) to KB and KBv200 cells was 1.37 and 1.42 μmol/L, respectively. IC 50 of H-21 (1-pyridylethylamine-3-methyl-6,8-dimethoxy-anthraquinone) was 13.0 and 17.9 μmol/L, IC 50 of H-25 (1-pyrrolylethylamine-3-methyl-6,8-dimethoxy-anthraquinone) was 8.5 and 11.7 μmol/L, IC 50 of H-28 (1-hydroxybutylamine-3-methyl-6,8-dimethoxy-anthraquinone) was 7.6 and 8.6 μmol/L. The IC 50 of them to MDR KBv200 cells was similar to that of them to the parental drug-sensitive KB cells (P>0.05). The generation of ROS increased obviously after the cells were incubated with them for 12 h, and the increase of ROS reached the peak treated for 24 h. The levels of ΔΨ m were time-dependently decreased after treating with four compounds for 12, 24, and 48 h. Conclusion The grouth of both MDR KBv200 cells and parental drug-sensitive KB cells were inhibited to the treatment of four anthraquinone derivative in vitro. The mechanism of their effects is associated with the increase of the cellular ROS level and the decrease of ΔΨ m.