ABC转运蛋白与临床白念珠菌耐药
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国家重点基础研究发展计划(“973计划”,2005CB523105).


ATP-binding-cassette transporters and fluconazole-resistance of clinical Candida albicans strains
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Supported by National Program on Key Basic Research Project of China ( “973” Program,2005CB523105).

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    摘要:

    目的:研究临床氟康唑耐药白念珠菌中CDR1和CDR2基因的表达情况与耐药性产生的关系,探寻临床白念珠菌耐药的分子机制。方法:微量液基稀释法测定氟康唑对白念珠菌临床分离株的最低抑菌浓度(minimal inhibitory concentration,MIC);实时定量PCR(RQ-PCR)方法检测主动外排泵基因CDR1和CDR2的表达水平;用罗丹明6G(Rhodamine 6G)外转运实验检测罗丹明6G在CDR1、CDR2高表达的白念珠菌中的外排情况。结果:氟康唑耐药组菌株CDR1和CDR2高表达发生率显著高于敏感组菌株;而在加入葡萄糖提供能量后,CDR1和CDR2蛋白发生高表达的耐药菌株罗丹明6G外排能力显著增加。部分耐药菌株中未见CDR1和CDR2的高表达。结论:白念珠菌临床分离株对氟康唑的耐药性与ABC转运蛋白过度表达相关;ABC转运蛋白的高表达可以使药物外排能力增加,从而导致耐药性产生。同时可能有其他机制参与临床白念珠菌耐药性的生成。

    Abstract:

    Objective:To explore the relationship between CDR1 and CDR2 expression and fluconazole-resistance in clinical Candida albicans strains, so as to explore the mechanism of drug-resistance in Candida albicans.Methods: The minimal inhibitory concentrations (MICs) of clinical Candida albicans strains to fluconazole were determined by broth microdilution method. The total RNA of fluconazole-susceptible and fluconazole-resistant Candida albicans strains were extracted,and the expression of the CDR1 and CDR2 genes was examined by real-time RT-PCR. The efflux of Rhodamine 6G was determined in the strains highly expressing CDR1 and CDR2 genes.Results: The incidences of CDR1 and (or) CDR2 overexpression in fluconazole-resistant strains were significantly higher than those in the fluconazole-sensitive stains. The efflux of Rhodamine 6G was significantly increased in the fluconazole-resistant strains when glucose was added, and overexpression of CDR1 and CDR2 were observed in theses strains. Some resistant strains showed no overexpression of CDR1 and CDR2.Conclusion: Fluconazole-resistance in clinical Candida albicans strains is related to the overexpression of ABC transporter proteins.Overexpression of ABC transporters can increase the efflux of drugs and therefore lead to drug-resistance. Other mechanisms may also participate in the development of drug resistance in the clinical Candida albicans strains.

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  • 收稿日期:2008-04-10
  • 最后修改日期:2008-06-03
  • 录用日期:2008-06-04
  • 在线发布日期: 2008-09-12
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