MIR1基因影响白念珠菌对唑类药物的敏感性

Effect of MIR1 gene on the sensitivity of Candida albicans to azoles

目的考察MIR1基因对白念珠菌对唑类药物敏感性的影响。方法以野生型白念珠菌、mir1Δ/Δ敲除菌和mir1Δ/MIR1回复菌为实验对象,应用斑点法考察MIR1基因敲除对抗真菌药物敏感性的影响,利用罗丹明6G外排实验考察白念珠菌MIR1基因敲除对药物外排能力的影响,利用JC-1考察MIR1基因敲除对线粒体膜电位的影响,利用BacTiter-Glo Reagent考察MIR1基因敲除对三磷酸腺甘(ATP)生成能力的影响。结果MIR1基因敲除使白念珠菌对唑类药物的敏感性提高,药物外排能力缺陷,线粒体膜电位有降低趋势,ATP生成能力缺陷。结论MIR1基因影响白念珠菌对唑类药物的敏感性,可能是通过影响ATP生成进而影响ABC转运蛋白的药物外排功能造成的。以白念珠菌MIR1为靶点,有望开发与唑类药物协同的新型抗真菌药物。

Objective To determine the role of MIR1 gene on the sensitivity of Candida albicans to azoles.Methods Wild-type Candida albicans,mir1Δ/Δmutant and mir1Δ/MIR1 mutant were used as experimental strains.Spot assay was conducted to determine the effect of MIR1 gene knockout on antifungal susceptibility.Rhodamine 6G was used to check the effect of MIR1 gene on drug efflux in Candida albicans.JC-1 was used to decide the effect of MIR1 gene on mitochondrial membrane potential.Finally,the effect of MIR1 gene on ATP production was examined by BacTiter-Glo Reagent.Results MIR1 gene knockout increased the sensitivity of Candida albicans to azoles,impaired drug efflux,decreased the mitochondrial membrane potential,and impaired the ability of ATP production.Conclusion MIR1 gene affects the sensitivity of Candida albicans to azoles,which may be caused by affecting ATP production and drug efflux mediated by ATP-binding cassette transporter.Targeting MIR1 gene in Candida albicans helps develop new antifungal drugs that synergize with azoles.