光滑念珠菌两性霉素B耐药株的体外诱导及其耐药分子机制  

Study on the induction of amphotericin B resistance in vitro in Candida glabrata and its corresponding molecular mechanism

作　　者：黄金梅[1] 叶宝国[1] 沈建箴[2] 吴淡森[2] 

机构地区：[1]福建医科大学附属漳州市医院血液科,福建漳州363000 [2]福建医科大学附属协和医院血液科,福建福州350001

出　　处：《中国医院药学杂志》2014年第9期688-692,共5页Chinese Journal of Hospital Pharmacy

基　　金：福建省科技厅产学研项目(编号:2011Y4002);福建卫生厅创新课题(编号:2009-cx-5);福省教育厅资助项目(编号:JA10129)

摘　　要：目的:体外诱导光滑念珠菌两性霉素B(AMB)耐药株,检测耐药株耐药基因CYP51A、CYP51B、CDR1、CDR2、MDR1、MDR2、Yap1、Sho1的表达情况,探讨其对AMB耐药的分子机制。方法:先以MTT微量液基稀释法检测光滑念珠菌对AMB的敏感性,再用AMB体外诱导其耐药株,并用实时荧光定量PCR法检测野生株与耐药株耐药基因的表达水平。结果:光滑念珠菌野生株有CYP51A、CYP51B、CDR1、CDR2、MDR1、MDR2、Yap1、Sho1基因低水平表达,而耐药株CYP51A、CYP51B、CDR1、CDR2、MDR1、MDR2、Yap1、Sho1基因表达水平明显升高,野生株与耐药株有显著差别(P<0.05)。结论:光滑念珠菌对AMB耐药与CYP51A、CYP51B、CDR1、CDR2、MDR1、MDR2、Yap1、Sho1基因过度表达有关,AMB可诱导麦角固醇合成过程中的关键酶基因、外排泵基因及信号传导相关基因过表达,从而产生耐药性。OBJECTIVE To induce the amphotericin B resistant strain of the Candida glabrata in vitro; to test the expres- sions of the drug-resistant genes and discuss the corresponding molecular mechanism. METHODS Amphotericin B susceptibil- ity was evaluated by a standard broth microdilution method. Then the resistant strain was induced with amphotericin B in vitro. The drug-resistant gene expression levels of the wild strain and the resistant strain were measured by real-time PCR method. RESULTS The wild strain had CYP51A, CYP51B, CDR1, CDR2, MDR1, MDR2, Yapl, Shol mRNA expression at low level in the Candida glabrata. But the mRNA expression level of the CYP51A, CYP51B, CDR1, CDR2, MDR1, MDR2, Yap1, Shol was higher in the amphotericin B resistant strain than in the wild strain （P〈0. 05）. CONCLUSION The excessive expression of the CYP51A, CYP51B, CDR1, CDR2, MDR1, MDR2, Yapl, Shol mRNA was associated with the resistance to amphotericin B in the Candida glabrata strains. Amphotericin B can induce the key enzyme gene in the synthesis of ergosterol , the efflux pumps genes and the signal transduction related genes expression leading to the resistance to the am- photericin B.

关 键 词：光滑念珠菌 两性霉素B 耐药 基因表达 

分 类 号：R969[医药卫生—药理学]