Aydin Merve, Unusan Nurhan, Sumlu Esra, Korucu Emine Nedime
Department of Medical Microbiology, Faculty of Medicine, KTO Karatay University, Konya 42020, Turkey.
Department of Nutrition and Dietetics, Faculty of Health Sciences, KTO Karatay University, Konya 42020, Turkey.
J Fungi (Basel). 2024 Oct 30;10(11):751. doi: 10.3390/jof10110751.
species, opportunistic pathogens that cause various infections, pose a significant threat due to their ability to form biofilms that resist antifungal treatments and immune responses. The increasing resistance of spp. and the limited availability of effective treatments have prompted the research of natural compounds as alternative therapies. This study assessed the antifungal properties of RA against species, focusing on its impact on s biofilms and the underlying mechanisms. The antifungal efficacy of RA was evaluated using the CLSI M27-A3 microdilution method on both fluconazole-susceptible and -resistant strains. Biofilm formation by was assessed through a crystal violet assay, while its antibiofilm activity was analyzed using an MTT assay and field emission scanning electron microscopy (FESEM). Gene expression related to biofilm formation was studied using quantitative real-time PCR (qRT-PCR), and statistical analysis was performed with an ANOVA. Among the 28 strains tested, RA exhibited minimum inhibitory concentration (MIC) values ranging from 160 to 1280 μg/mL. At a 640 μg/mL concentration, it significantly reduced the expression of genes associated with adhesion (, , and ), hyphal development ( and ), and hyphal cAMP-dependent protein kinase regulators (, , and ) in RAS1-cAMP-EFG1 pathway ( < 0.05). FESEM analysis revealed a reduction in hyphal networks and disruptions on the cell surface. Our study is the first to demonstrate the effects of RA on adhesion, hyphae development, and biofilm formation through gene expression analysis with findings supported by FESEM. This approach distinguishes our study from previous studies on the effect of RA on . However, the high MIC values of RA limit its antifungal potential. Therefore, more extensive research using innovative methods is required to increase the antifungal effect of RA.
某些物种作为机会性致病菌会引发各种感染，因其能够形成抵抗抗真菌治疗和免疫反应的生物膜，从而构成重大威胁。[物种名称]的耐药性不断增加以及有效治疗方法的有限可用性促使人们研究天然化合物作为替代疗法。本研究评估了RA对[物种名称]的抗真菌特性，重点关注其对[物种名称]生物膜的影响及潜在机制。采用CLSI M27 - A3微量稀释法在氟康唑敏感和耐药菌株上评估RA的抗真菌疗效。通过结晶紫测定法评估[物种名称]的生物膜形成，同时使用MTT测定法和场发射扫描电子显微镜（FESEM）分析其抗生物膜活性。使用定量实时PCR（qRT - PCR）研究与生物膜形成相关的基因表达，并采用方差分析进行统计分析。在所测试的28株[菌株名称]菌株中，RA的最低抑菌浓度（MIC）值范围为160至1280μg/mL。在640μg/mL浓度下，它显著降低了RAS1 - cAMP - EFG1途径中与黏附（[基因名称1]、[基因名称2]和[基因名称3]）、菌丝发育（[基因名称4]和[基因名称5]）以及菌丝cAMP依赖性蛋白激酶调节因子（[基因名称6]、[基因名称7]和[基因名称8]）相关的基因表达（P < 0.05）。FESEM分析显示菌丝网络减少且细胞表面受到破坏。我们的研究首次通过基因表达分析证明了RA对[物种名称]黏附、菌丝发育和生物膜形成的影响，FESEM的结果支持了这些发现。这种方法使我们的研究有别于先前关于RA对[物种名称]影响的研究。然而，RA的高MIC值限制了其抗真菌潜力。因此，需要使用创新方法进行更广泛的研究以提高RA的抗真菌效果。
