| 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。 |
| 关键词: 白念珠菌 氟康唑 真菌耐药性 ABC转运蛋白 罗丹明6G |
| DOI：10.3724/SP.J.1008.2008.01038 |
| 投稿时间：2008-04-10修订日期：2008-06-03 |
| 基金项目:国家重点基础研究发展计划(“973计划”,2005CB523105). |
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| ATP-binding-cassette transporters and fluconazole-resistance of clinical Candida albicans strains |
| SUN Wei-wei1,GAO Ping-hui1, WANG Ying2, JIANG Yuan-ying1* |
| (1.Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China 2.Department of Dermatology, Changhai Hospital, Second Military Medical University, Shanghai 200433) |
| 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. |
| Key words: Candida albicans fluconazole fungal drug resistance ABC transporter proteins Rohdamine 6G |
