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Effects of fluconazole on Candida glabrata biofilms and its relationship with ABC transporter gene expression

机译:氟康唑对光滑念珠菌生物膜的影响及其与ABC转运蛋白基因表达的关系

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摘要

Candida glabrata has emerged as the second most prevalent fungal pathogen and its ability to form biofilms has been considered one of the most important virulence factors, since biofilms present a high tolerance to antifungal agents used in fungal infection treatment. The mechanisms of biofilm tolerance to antifungal agents remain poorly understood. Thus, the aim of this study was to evaluate the effects of fluconazole (FLU) on the formation and control of C. glabrata biofilms and its relation with the expression of genes encoding for ABC transporters, CDR1, SNQ2, and PDR1. For that, minimal inhibitory concentration values for seven C. glabrata strains were determined and the effect of FLU against C. glabrata biofilms was evaluated by total biomass quantification and viable cell enumeration. Matrices from biofilms were analyzed in terms of protein, carbohydrate and DNA content. ABC transporter gene expression was analyzed for quantitative real-time PCR. In addition to the high amounts of proteins and carbohydrates detected in the extracellular matrices in the presence of FLU, this work showed that the overexpression of efflux pumps is a possible mechanism of biofilm tolerance to FLU and this phenomenon alters the structure of C. glabrata biofilms by creating cell clusters.
机译:光滑念珠菌已成为第二大流行的真菌病原体,其形成生物膜的能力被认为是最重要的毒力因子之一,因为生物膜对真菌感染治疗中使用的抗真菌剂具有很高的耐受性。生物膜抗真菌剂耐受性的机制仍知之甚少。因此,本研究的目的是评估氟康唑(FLU)对光滑小球藻生物膜形成和控制的作用及其与编码ABC转运蛋白,CDR1,SNQ2和PDR1的基因表达的关系。为此,确定了七种光滑毛霉菌菌株的最小抑菌浓度值,并通过总生物量定量和活细胞计数评估了FLU对光滑毛霉菌生物膜的影响。对生物膜中的基质进行了蛋白质,碳水化合物和DNA含量分析。分析ABC转运蛋白基因表达以用于定量实时PCR。除了在存在FLU的情况下在细胞外基质中检测到大量蛋白质和碳水化合物外,这项工作还表明外排泵的过表达是生物膜对FLU的耐受性的一种可能机制,并且这种现象改变了光滑念珠菌生物膜的结构。通过创建单元簇。

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