Silymarin exerts antifungal effects via membrane-targeted mode of action by increasing permeability and inducing oxidative stress
Silymarin, which is derived from the seeds of Silybum marianum, has been widely used to prevent and treat liver disorders. It is also consumed as a dietary supplement to improve liver function, as it does not exhibit any toxic effects in humans. Recently, silymarin has been reported to show antimicrobial effects against various pathogenic microorganisms, but the mode of action remains unknown. Thus, we investigated the antifungal activity of silymarin and aimed to determine the underlying mechanism. Initially, a propidium iodide assay was carried out; the results indicated that silymarin induced injury to the fungal plasma membrane. Subsequently, large unilamellar vesicles encapsulating calcein and fluorescein isothiocyanate-labeled dextrans (FDs) 4, 10, and 20 were prepared to analyze whether silymarin affects an artificial membrane model. The results indicated that silymarin increased membrane permeability by disturbing the membrane structure, thereby allowing free access to molecules smaller than FD20 (approximately 3.3nm). The accumulation of reactive oxygen species (ROS) results in deleterious effects to various cellular components. In particular, ROS easily react with the membrane lipids and induce lipid peroxidation, which increases membrane permeability and disturbs hydrophobic phospholipids. Using 2',7'-dichlorodihydrofluorescein diacetate and thiobarbituric acid, we confirmed that silymarin induced harmful effects on the plasma membrane. Membrane depolarization and K + leakage, which were associated with an increase in membrane permeability, were also observed in Candida albicans cells. An assay using 1,6-diphenyl-1,3,5-hexatriene showed that silymarin decreased membrane fluidity. Taken together, we suggest that silymarin exerts its antifungal activity by targeting the C. albicans plasma membrane.
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