Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata - Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Mar del Plata, Argentina.
Laboratório de Fisiologia e Bioquímica de Microrganismos, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, Brazil.
J Appl Microbiol. 2022 Jun;132(6):4310-4320. doi: 10.1111/jam.15544. Epub 2022 Apr 5.
This study aimed to evaluate the combined effect of a mannose-binding lectin Helja with fluconazole (FLC) on Candida albicans and to get insights about the joint action mechanism.
The fungal growth was assessed following the optical density at 630 nm. Fungal cell morphology and nucleus integrity were analysed by flow cytometry and confocal laser scanning microscopy using Calcofluor White (CFW) and 4',6-diamidino-2-phenylindole (DAPI) staining respectively. The basis of Helja + FLC action on cell wall and plasma membrane was analysed using perturbing agents. The Helja + FLC combination exhibited an inhibitory effect of fungal growth about three times greater than the sum of both compounds separately and inhibited fungal morphological plasticity, an important virulence attribute associated with drug resistance. Cells treated with Helja + FLC showed morphological changes, nucleus disintegration and formation of multimera structures, leading to cell collapse.
Our findings indicate that the Helja + FLC combination exhibited a potent antifungal activity based on their simultaneous action on different microbial cell targets.
The combination of a natural protein with conventional drugs might be helpful for the design of effective therapeutic strategies against Candida, contributing to minimize the development of drug resistance and host cell toxicity.
本研究旨在评估甘露聚糖结合凝集素 Helja 与氟康唑（FLC）联合对白色念珠菌的协同作用，并深入了解联合作用机制。
通过在 630nm 处测量吸光度来评估真菌生长。使用流式细胞术和共聚焦激光扫描显微镜，分别用 Calcofluor White（CFW）和 4'，6-二脒基-2-苯基吲哚（DAPI）染色分析真菌细胞形态和核完整性。使用干扰剂分析 Helja+FLC 对细胞壁和质膜的作用基础。Helja+FLC 联合用药对真菌生长的抑制作用比两种化合物单独使用的总和强约三倍，且抑制了真菌形态可塑性，这是与耐药性相关的一个重要毒力属性。用 Helja+FLC 处理的细胞表现出形态变化、核解体和多聚体结构的形成，导致细胞崩溃。
我们的研究结果表明，Helja+FLC 联合用药基于其对不同微生物细胞靶标的同时作用，表现出了强大的抗真菌活性。
天然蛋白与常规药物的联合使用可能有助于设计针对白色念珠菌的有效治疗策略，有助于减少耐药性的发展和宿主细胞毒性。
