Department of Microbiology, Faculty of Science, Qom Branch, Islamic Azad University, Qom, Iran.
Nuclear Medicine Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Biotechnol Prog. 2022 Jan;38(1):e3206. doi: 10.1002/btpr.3206. Epub 2021 Sep 15.
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.
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。