华西口腔医学杂志

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白假丝酵母菌药物流出泵基因在生物膜中表达的动态研究

亓庆国1, 2, 周学东2, 杨德琴3, 肖晓蓉2, 刘正1   

  1. 1.上海交通大学医学院附属第九人民医院口腔科, 上海200025; 2.口腔生物医学工程教育部重点实验室, 四川大学, 四川成都610041; 3.遵义医学院附属口腔医院口腔内科, 贵州遵义563003
  • 收稿日期:2007-08-25 修回日期:2007-08-25 出版日期:2007-08-20 发布日期:2007-08-20
  • 通讯作者: 刘正,Tel:021- 63135412
  • 作者简介:亓庆国(1972-),男,山东人,副教授,博士
  • 基金资助:

    国家自然科学基金资助项目(30400498)

Dynamic study on Saccharomyces albicans drug efflux pumps gene expr ession dur ing the biofilm formation

QI Qing- guo1,2, ZHOU Xue- dong2, YANG De- qin3, XIAO Xiao- rong2, LIU Zheng1   

  1. 1. Dept. of Dentistry, The Ninth Hospital Medical College of Shanghai, Jiaotong University, Shanghai 200025, China; 2. Key. Laboratory of Oral Biomedical Engineering of Ministry of Education, Sichuan University, Chengdu 610041, China; 3. Dept. of Oral Medicine, Stomatological Hospital of Zunyi Medical College, Zunyi 563003, China
  • Received:2007-08-25 Revised:2007-08-25 Online:2007-08-20 Published:2007-08-20
  • Contact: LIU Zheng,Tel:021- 63135412

摘要:

目的动态观察白假丝酵母菌生物膜形成过程中耐药性产生相关的药物流出泵基因的表达和耐药性的变化情况,并分析该类基因的表达与生物膜耐药性产生的相关性。方法利用人工口腔恒化器形成白假丝酵母菌生物膜结构,收集生物膜形成不同阶段的生物膜态菌细胞,半定量RT- PCR技术分析菌细胞mRNA在生物膜成熟过程中的动态变化;96孔板XTT减低法获得生物膜耐药性的变化情况。结果耐药性随着生物膜的成熟不断增强;与早期生物膜相比,成熟生物膜菌株的药物流出泵基因CDR1表达上调,而MDR1基因的表达在生物膜的整个成熟过程中基本维持不变。结论生物膜成熟程度与耐药性增强有明显正相关性;药物流出泵基因的表达上调与生物膜耐药性的产生有一定关系但改变不一致;在生物膜成熟过程中,CDR1表达水平改变活跃,而MDR1基因相对稳定。

关键词: 白假丝酵母菌, 生物膜, 药物耐受性

Abstract:

Objective To observe the drug resistance and drug efflux pumps gene mRNA of Saccharomyces albicans, including CDR1 gene and MDR1 gene, at different stage of biofilm formation in chemostat, furthermore to analysis the relationship between the drug efflux pump gene expression and the biofilm related drug resistance. Methods To form the mature biofilm in vitro in chemostat, then collect the biofilm strains at different development stages(2, 12, 24, 48 h)to semi- quantified mRNA amount of CDR1 gene and MDR1 gene by one step RT- PCR method. Using XTT reduction method to test the dynamic change of Saccharomyces albicans drug resistance in biofilm. Results Antifungal resistance of biofilm- grown cells increased conjunction with the biofilm maturation. Compared with earth stage of biofiom strains, the amount of CDR1 mRNA gene in mature biofilm strains increased, while MDR1 gene did not. Conclusion There is positive correlation between drug resistance and biofilm maturation of Saccharomyces albicans. Biofilm related drug resistance appears to be partially associated with the upregulation of drug efflux pumps, although the variation is not shown coincidence. During the biofilm formation, CDR1 gene expression is actively up- regulated, but MDR1 gene expression is stable.

Key words: Saccharomyces albicans, biofilm, drug resistance