芯片毛细管电泳结合光谱法对阿霉素和谷胱甘肽相互作用的研究

Interaction of doxorubicin and glutathione by microchip capillary electrophoresis coupled with spectroscopic methods

目的:通过考察谷胱甘肽与阿霉素的相互作用,探讨谷胱甘肽在阿霉素耐药机制中的作用。方法:用芯片毛细管电泳激光诱导荧光技术,萘-2,3-二甲醛在线衍生法考察阿霉素和谷胱甘肽的相互反应情况;用紫外分光光度法、荧光分光光度法、简化的Hummel-Dreyer芯片毛细管电泳法考察阿霉素与还原性、氧化型谷胱甘肽的亲和作用。结果:还原型的谷胱甘肽不能与阿霉素直接结合,氧化型谷胱甘肽与阿霉素有弱结合;还原型的谷胱甘肽自身不稳定,容易被氧化,其氧化产物之一,磺酸化物与阿霉素可以发生亲和作用。结论:谷胱甘肽自身对阿霉素的耐药作用不大。

Objective:To investigate the interaction between glutathione and doxorubicin,and to explain the mechanism of doxorubicin resistance.Methods:The reaction between doxorubicin and glutathione was detected by a microchip capillary electrophoresis laser-induced fluorescence detector with an on-line derivatization technique by naphthalene-2,3-dicarbaldehyde(NDA).The affinity of doxorubicin with glutathione and oxidized glutathione were investigated separately by UV spectrophotometry,fluorescence spectrophotometry and simplified Hummel-Dreyer chip capillary electrophoresis.Results:Reduced glutathione couldn’t bind directly to doxorubicin.Oxidized glutathione at a high concentration had a weak association with doxorubicin.Reduced glutathione was not stable and was easily oxidized.The oxidation product of glutathione,glutathione-sulfonate,had an affinity with doxorubicin.Conclusion:Glutathione has little effect on the resistance mechanism of doxorubicin.