中国真菌学杂志
中國真菌學雜誌
중국진균학잡지
CHINESE JOURNAL OF MYCOLOGY
2015年
2期
75-79
,共5页
许洪涛%唐若愚%屈莺歌%王晓娟%姜远英%曹永兵%颜天华
許洪濤%唐若愚%屈鶯歌%王曉娟%薑遠英%曹永兵%顏天華
허홍도%당약우%굴앵가%왕효연%강원영%조영병%안천화
白念珠菌%转录因子%药物敏感性%多药耐药基因
白唸珠菌%轉錄因子%藥物敏感性%多藥耐藥基因
백념주균%전록인자%약물민감성%다약내약기인
Candida albicans%transcription factors%drug susceptibility%multidrug resistance genes
目的：利用基因缺失菌库研究转录因子敲除后对白念珠菌药物敏感性的影响，并利用药物敏感性差异菌株初步考察可能的耐药性调控机制。方法微量液基稀释法测定最低抑菌浓度（minimal inhibitory concentration，MIC）；点板法（spot assay）和生长曲线法验证菌株对氟康唑的敏感性；实时定量 PCR （RT?PCR）法检测药物敏感性差异菌株中多药耐药基因 CDR1和 MDR1的表达，并通过罗丹明6G 外排实验测定外排能力。结果亲本菌 SN250对氟康唑表现为耐药，多药耐药基因 CDR1和 MDR1高表达，MIC80大于16μg／ mL，大部分转录因子缺失菌与亲本菌 SN250表现出相同的耐药性，但转录因子 RPN4缺失菌株对氟康唑的敏感性升高，MIC80降为0．5μg／ mL；多药耐药基因 CDR1和 MDR1的表达均降低，20 min 和40 min 时对罗丹明6G 的外排能力降低。结论转录因子 RPN4有可能通过促进多药耐药基因 CDR1和 MDR1表达和菌株外排能力而降低对药物的敏感性，但相关机制有待进一步深入研究。
目的：利用基因缺失菌庫研究轉錄因子敲除後對白唸珠菌藥物敏感性的影響，併利用藥物敏感性差異菌株初步攷察可能的耐藥性調控機製。方法微量液基稀釋法測定最低抑菌濃度（minimal inhibitory concentration，MIC）；點闆法（spot assay）和生長麯線法驗證菌株對氟康唑的敏感性；實時定量 PCR （RT?PCR）法檢測藥物敏感性差異菌株中多藥耐藥基因 CDR1和 MDR1的錶達，併通過囉丹明6G 外排實驗測定外排能力。結果親本菌 SN250對氟康唑錶現為耐藥，多藥耐藥基因 CDR1和 MDR1高錶達，MIC80大于16μg／ mL，大部分轉錄因子缺失菌與親本菌 SN250錶現齣相同的耐藥性，但轉錄因子 RPN4缺失菌株對氟康唑的敏感性升高，MIC80降為0．5μg／ mL；多藥耐藥基因 CDR1和 MDR1的錶達均降低，20 min 和40 min 時對囉丹明6G 的外排能力降低。結論轉錄因子 RPN4有可能通過促進多藥耐藥基因 CDR1和 MDR1錶達和菌株外排能力而降低對藥物的敏感性，但相關機製有待進一步深入研究。
목적：이용기인결실균고연구전록인자고제후대백념주균약물민감성적영향，병이용약물민감성차이균주초보고찰가능적내약성조공궤제。방법미량액기희석법측정최저억균농도（minimal inhibitory concentration，MIC）；점판법（spot assay）화생장곡선법험증균주대불강서적민감성；실시정량 PCR （RT?PCR）법검측약물민감성차이균주중다약내약기인 CDR1화 MDR1적표체，병통과라단명6G 외배실험측정외배능력。결과친본균 SN250대불강서표현위내약，다약내약기인 CDR1화 MDR1고표체，MIC80대우16μg／ mL，대부분전록인자결실균여친본균 SN250표현출상동적내약성，단전록인자 RPN4결실균주대불강서적민감성승고，MIC80강위0．5μg／ mL；다약내약기인 CDR1화 MDR1적표체균강저，20 min 화40 min 시대라단명6G 적외배능력강저。결론전록인자 RPN4유가능통과촉진다약내약기인 CDR1화 MDR1표체화균주외배능력이강저대약물적민감성，단상관궤제유대진일보심입연구。
Objective To study the influence on the drug susceptibility of Candida albicans after transcription factor RPN4 was knocked out and the mechanism how this transcription factor regulates the drug susceptibility in Candida albicans.Methods The broth microdilution method was used to determine the minimal inhibitory concentrations (MICs) of 30 strains transcription factor knocked out Candida albicans .The drug susceptibility was reassured by spot assay and growth curve.The total RNA of parental strain SN250 and susceptible strains were extracted and the expression of multidrug resistance genes CDR1 and MDR1 was determined by real?time PCR,and the pump ability was examined by the ability of pump out rhodamine 6G.Results Compaired to the parental strain SN250,transcription factor RPN4 knocked out strain was more susceptible to fluconazole.The expression of CDR1 and MDR1 was lower than SN250 and the pump ability was also weaker than SN250.Conclusion The knockout of transcription factor PRN4 re?sults susceptible to fluconazole in Candida albicans,and this transcription factor may be involved in the regulation of drug susceptibil?ity in Candida albicans.
