穿心莲内酯联合氟康唑抗耐药白假丝酵母菌作用机制研究  

Mechanisms of the combination effects of Andrographolide and Fluconazole against fluconazole-resistant Candida albicans

作　　者：施高翔[1,2] 严园园[1,2] 邵菁[1,2] 陆克乔 张梦翔[1,2] 汪天明[1,2] 汪长中[1,2] 

机构地区：[1]安徽中医药大学中西医结合临床学院,安徽合肥230038 [2]安徽省中医药科学院中西医结合研究所,安徽合肥230038

出　　处：《中国微生态学杂志》2014年第3期258-264,共7页Chinese Journal of Microecology

基　　金：国家自然科学基金项目(81073127)

摘　　要：目的探讨穿心莲内酯联合氟康唑抗耐药白假丝酵母菌作用及其机制。方法采用微量稀释法检测穿心莲内酯(AG)及联合氟康唑(FLC)对耐药白假丝酵母菌的MIC;采用罗丹明6G(Rh 6G)检测AG对耐药白假丝酵母菌CDR外排功能的影响;利用罗丹明123评估AG对耐药白假丝酵母菌MDR外排功能的影响;采用二氢罗丹明检测AG单用及联合FLC对耐药白假丝酵母菌活性氧(ROS)的影响;采用实时荧光定量PCR(qRT-PCR)检测AG联合FLC对耐药白假丝酵母菌外排泵相关基因CDR1、CDR2和MDR1表达的影响。结果 AG联合FLC抗耐药白假丝酵母菌呈相加作用;AG对CDR外排功能无影响;AG可抑制MDR外排功能;AG联合FLC能显著提高耐药白假丝酵母菌细胞ROS水平;AG与FLC联合作用于耐药白假丝酵母菌可下调CDR1和MDR1的表达量,上调CDR2的表达量。结论 AG联合FLC抗耐药白假丝酵母菌具有相加作用,其机制可能与抑制外排泵及相关基因表达,提高胞内ROS水平有关。Objective To investigate effects of Andrographolide combination with fluconazole on fluconazole-resist- ance in Candida albicans and its mechanism. Methods The antifungal activities of AG and FLC and determine minimum inhibitory concentration （MIC） were evaluated by mierodilution method on a fluconazole-resistant Candi- da albicans isolate 215 ; The efflux of Rhodamine 6G and Rhodamine 123 were observed in Candida albicans mem- brane; The levels of reactive oxygen species （ROS） with DHR staining was measured by flow eytometry; Efflux-re- [ated genes expression of CDR1 ,CDR2 and MDR1 were inspected by the real-time fluorescence quantification PCR. Results The antifungal effects showed significantly enhanced by AG combination with FLC, and the combination showed clear collaboration; AG could not affect the efflux of rhodamine 6G, and it dramatically reduce efflux of rhodamine 123 and promote the production of intraeelluar ROS with dose-dependent mode; And expressions of CDR1 and MDR1 were significantly downregulated, while the expression of CDR2 was upregulated by qRT-PCR.Conclusion The study indicates that AG can enhance the antifungal activity of FLC by inhibiting the ef？ ux of FLC from C. albicans and raising the levels of ROS.

关 键 词：穿心莲内酯 氟康唑 耐药白假丝酵母菌 ROS 外排泵 

分 类 号：R379.4[医药卫生—病原生物学]