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生物合成硒纳米颗粒对烟曲霉和黄曲霉中 CYP51A 和 HSP90 抗真菌耐药基因表达的影响。

The effect of biosynthesized selenium nanoparticles on the expression of CYP51A and HSP90 antifungal resistance genes in Aspergillus fumigatus and Aspergillus flavus.

机构信息

Department of Microbiology, Faculty of Science, Qom Branch, Islamic Azad University, Qom, Iran.

Nuclear Medicine Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.

出版信息

Biotechnol Prog. 2022 Jan;38(1):e3206. doi: 10.1002/btpr.3206. Epub 2021 Sep 15.

Abstract

The application of biological nanoparticles (NPs) can be considered as a way to overcome the problem of antifungal resistance in pathogenic fungi. This study takes a new approach to biosynthesized NPs influence on the expression of CYP51A and HSP90 antifungal resistance genes in Aspergillus fumigatus and A. flavus, and comparison with antifungal agents. Selenium NPs (Se-NPs) were biosynthesized using Aspergillus strains and their production was proved by several methods including, UV-Vis, XRD, FTIR, FESEM, and EDX techniques. The minimum inhibitory concentrations (MICs) of Aspergillus strains were determined using the CLSI M38-A2 broth microdilution method. The differences in expression levels of CYP51A and HSP90 genes were examined between untreated and treated of A. fumigatus and A. flavus using itraconazole and amphotericin B and biosynthesized Se-NPs through real-time PCR. After confirming the results of NPs synthesis, the MIC of itraconazole and amphotericin B against A. fumigatus and A. flavus was 4 μg/ml. Based on the real-time PCR results, the obtained ∆∆CTs for these strains were -0.18, -1.46, and -1.14. Whereas the MIC values for treated samples with Se-NPs have decreased to 0.5 μg/ml, and the ∆∆CTs for these were -0.25, -1.76, and -1.68. The expression of CYP51A and HSP90 genes was significantly down-regulated through the use of Se-NPs against A. fumigatus and A. flavus.

摘要

生物纳米粒子(NPs)的应用可以被视为克服致病真菌中抗真菌耐药性问题的一种方法。本研究采用新方法研究生物合成 NPs 对烟曲霉和黄曲霉 CYP51A 和 HSP90 抗真菌耐药基因表达的影响,并与抗真菌药物进行比较。使用曲霉属菌株合成硒纳米粒子(Se-NPs),并通过紫外-可见分光光度法、X 射线衍射(XRD)、傅里叶变换红外光谱(FTIR)、场发射扫描电子显微镜(FESEM)和能谱(EDX)等多种方法证明其生产。采用 CLSI M38-A2 肉汤微量稀释法测定曲霉属菌株的最小抑菌浓度(MIC)。采用实时 PCR 检测未处理和处理的烟曲霉和黄曲霉中 CYP51A 和 HSP90 基因的表达水平。在确认 NPs 合成结果后,测定烟曲霉和黄曲霉对伊曲康唑和两性霉素 B 的 MIC 分别为 4μg/ml。根据实时 PCR 结果,这些菌株的 ∆∆CT 值分别为-0.18、-1.46 和-1.14。而用 Se-NPs 处理后的样本 MIC 值降低至 0.5μg/ml,这些菌株的 ∆∆CT 值分别为-0.25、-1.76 和-1.68。Se-NPs 对烟曲霉和黄曲霉的使用显著下调了 CYP51A 和 HSP90 基因的表达。

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