Antifungal property of dihydrodehydrodiconiferyl alcohol 9′-O-β-d-glucoside and its pore-forming action in plasma membrane of Candida albicans

Abstract

The aims of this study wereto investigate the antifungal activity as a bioactive property of dihydrodehydro-diconiferyl alcohol 9'-O-3-D-glucoside (DDDC9G) and the mode of action(s) involved in its effect. Antifungal susceptibility testing showed that DDDC9G possessed potent antifungal activities toward various fungal strains with almost no hemolytic effect. To understand the antifungal mechanism(s) of DDDC9G, we conducted the following experiments in this study using Candida albicans. Fluorescence experiments using the probe 1,6-eiphenyl-1, 3, 5-hexatriene (DPH) and propidium iodide suggested that DDDC9G perturbed the fungal plasma membrane. Consecutively, the analysis of the transmembrane electrical potential (DeltaPsi) with 3, 3'-dipropylthiadicarbocyanine iodide [DiSC3(5)] and bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)] indicated that DDDC9G induced membrane-depolarization. Furthermore, model membrane studies were performed wiith rhodamine-labeled giant unilamellar vesicles (GUVs), calcein encapsulating large unilamellar vesicles (ILUVs), and FITC-dextran (FD) loaded LUVs. These results demonstrated that the antifungal effects of DDDC9G upon the fungal plasma membrane were through the formation of pores with the radii between 0.74 nm and 1.4 nm. Finally, in three dimensional (3D) flow cytometric contour plots, a reduced cell size was observed as a result of osmolarity changes from DDDC9G-induced structural and functional membrane damages.Therefore, the present study suggests that DDDC9G exerts its antifungal effect by damaging the membrane through pore formation in the fungal plasma membrane.

An Antifungal Mechanism of Protolichesterinic Acid from the Lichen Usnea albopunctata Lies in the Accumulation of Intracellular ROS and Mitochondria-Mediated Cell Death Due to Apoptosis in Candida tropicalis

“…After entering, PI intercalates between the guanine and the cytosine pair of DNA or with a stoichiometry of one unit dye per 3–5 base pairs. After DNA binding, the red fluorescent color of PI enhances more than 20-fold (Suzuki et al, 1997; Choi et al, 2012). From our study, we showed the distribution of the PI inside the C. tropicalis through the injured plasma membrane, and finally establish the cell membrane active mechanisms of protolichesterinic acid in Candida cells.…”

“…From our study, we showed the distribution of the PI inside the C. tropicalis through the injured plasma membrane, and finally establish the cell membrane active mechanisms of protolichesterinic acid in Candida cells. Maintaining the cytoplasmic membrane integrity is essential to various important functions of microorganisms, including the formation of gradient and discriminating permeability, cellular energetic, synthesis of various biopolymers, and other key virulence determinants (Choi et al, 2012). In this admiration, any changes in the physical state of the cell membrane induced by protolichesterinic acid may hinder with one or more functions directly or indirectly.…”

An antifungal mechanism of curcumin lies in membrane‐targeted action within Candida albicans

“…To further elucidate the membrane-active mechanisms, we used calcein-containing GUVs composed of PC/PE/PI/ergosterol (5:4:1:2, wt/wt/wt/wt; ref. 34), which mimic the membranes of C. albicans and were prepared using the electroformation method (15). GUVs have become a useful model for imitating biological membranes (35), and morphological changes in the artificial membrane model allow the study of structural and physical changes occurring during interactions with antimicrobial agents (36).…”

Blad-Containing Oligomer Fungicidal Activity on Human Pathogenic Yeasts. From the Outside to the Inside of the Target Cell

“…Damages at the cell membrane level, measured by propidium iodide uptake, are usually indicators of cell death (Simonin et al, 2007; Choi et al, 2012), but it is also possible that some damaged cells retain the ability to recover after a short incubation (Davey and Hexley, 2011). Propidium iodide may also be used as an indicator of cell leakage, or cell permeability (Piotrowski et al, 2015).…”