Journal of Advanced Research ( IF 11.4 ) Pub Date : 2021-06-17 , DOI: 10.1016/j.jare.2021.06.016 Xueyang Jin 1 , Ming Zhang 2 , Jinghui Lu 1 , Ximeng Duan 1 , Jinyao Chen 1 , Yue Liu 1 , Wenqiang Chang 1 , Hongxiang Lou 1
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Introduction
The increasing morbidity of fungal infections and the prevalence of drug resistance highlighted the discovery of novel antifungal agents and investigation of their modes of action. Iron chelators have been used to treat superficial fungal infections or potentiate the efficacy of certain antifungal drugs. Hinokitiol exhibits potent antifungal activity and iron-chelating ability. However, their relationships have not been established.
Objectives
This study aims to explore the selectivity of hinokitiol against fungal cells and mammalian cells and determine the role of iron-chelating for the antifungal activity of hinokitiol.
Methods
Iron probe FeRhonox-1 was used to determine intracellular Fe2+ content. 5-Cyano-2,3-ditolyl tetrazolium chloride probe and Cell Counting Kit-8 were used to detect the mitochondrial respiratory activities. Quantitative real-time PCR and rescue experiments were performed to determine the effect of iron on the antifungal activity of hinokitiol. The effects of hinokitiol on fungal mitochondria were further evaluated using reactive oxygen species probes and several commercial Assay Kits. The ability of hinokitiol to induce resistance in Candida species was carried out using a serial passage method. The in vivo therapeutic effect of hinokitiol was evaluated using Galleria mellonella as an infectious model.
Results
Hinokitiol was effective against a panel of Candida strains with multiple azole-resistant mechanisms and persistently inhibited Candida albicans growth. Mechanism investigations revealed that hinokitiol chelated fungal intracellular iron and inhibited the respiration of fungal cells but had minor effects on mammalian cells. Hinokitiol further inhibited the activities of mitochondrial respiratory chain complexes I and II and reduced mitochondrial membrane potential, thereby decreasing intracellular ATP synthesis and increasing detrimental intracellular reductive stress. Moreover, hinokitiol exhibited low potential for inducing resistance in several Candida species and greatly improved the survival of Candida-infected Galleria mellonella.
Conclusions
These findings suggested the potential application of hinokitiol as an iron chelator to treat fungal infections.
中文翻译：
桧醇螯合细胞内铁，通过干扰线粒体呼吸来延缓真菌生长
介绍
真菌感染发病率的增加和耐药性的普遍存在突出了新型抗真菌剂的发现和对其作用方式的研究。铁螯合剂已被用于治疗浅表真菌感染或增强某些抗真菌药物的功效。桧木醇表现出有效的抗真菌活性和铁螯合能力。但是，他们的关系尚未确定。
目标
本研究旨在探索桧醇对真菌细胞和哺乳动物细胞的选择性，并确定铁螯合对桧醇抗真菌活性的作用。
方法
铁探针FeRhonox-1用于测定细胞内Fe 2+含量。5-Cyano-2,3-ditolyl tetrazolium chloride 探针和 Cell Counting Kit-8 用于检测线粒体呼吸活动。进行定量实时 PCR 和拯救实验以确定铁对扁柏醇抗真菌活性的影响。使用活性氧探针和几种商业化验试剂盒进一步评估了扁柏醇对真菌线粒体的影响。桧醇在念珠菌属中诱导抗性的能力是使用连续传代法进行的。使用大花菌作为感染模型评估了扁柏醇的体内治疗效果。
结果
桧醇对一组具有多种唑类抗性机制的念珠菌菌株有效，并持续抑制白色念珠菌的生长。机制研究表明，桧醇螯合真菌细胞内的铁并抑制真菌细胞的呼吸作用，但对哺乳动物细胞的影响较小。Hinokitol 进一步抑制线粒体呼吸链复合物 I 和 II 的活性并降低线粒体膜电位，从而降低细胞内 ATP 合成并增加有害的细胞内还原应激。此外，桧醇在几种念珠菌中表现出较低的诱导抗药性的潜力，并大大提高了念珠菌感染的存活率梅洛内拉。
结论
这些发现表明桧醇作为铁螯合剂治疗真菌感染的潜在应用。
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