Effects of Different Concentrations of AmB on the Unsaturated Phospholipid-Cholesterol Membrane Using the Langmuir Monolayer and Liposome Models.

Amphotericin B (AmB) causes toxicity to the erythrocyte membrane, leading to hemolysis, which limits the clinically effective dose for AmB intravenous therapy in invasive fungal infections. The molecular mechanism by which AmB adheres to the membrane of erythrocytes is the key factor in causing AmB to be toxic to the membrane of erythrocytes, but it is not yet fully understood; the mechanism by which AmB adheres to the liquid microdomains with higher fluidity formed by cholesterol and unsaturated phospholipids remains especially unclear. This study examined the adsorption of AmB at different concentrations, 5, 45, 85, and 125 μg/mL, on unsaturated phospholipid membranes containing 50 mol% cholesterol. The thermodynamic properties and structure of DOPC monolayers and DOPC/cholesterol mixed monolayers at different concentrations of AmB have been investigated using the Langmuir monolayer model and the BAM method. The impact of varying concentrations of AmB on the hydrophilic and hydrophobic domains of the DOPC bilayers and the DOPC/cholesterol mixed bilayers have also been discussed using large unilamellar vesicle liposomes and fluorescence techniques. It is shown that for AmB concentrations greater than 5 μg/mL, with an increase in AmB's concentration, the reorganization time for the DOPC/cholesterol monolayer increases, and the elastic modulus of the DOPC/cholesterol mixed monolayer decreases. In particular, when AmB's concentration is higher than 85 μg/mL, the liquid-condensed phase domains on the DOPC/cholesterol monolayer reduce significantly and the liquid-expanded phase domain enlarges from the BAM images. When the AmB concentration reaches 5 μg/mL, the disorder of the hydrophobic and hydrophilic domains of the DOPC/cholesterol bilayer increases as the AmB concentration increases. The way in which AmB interacts with the DOPC/cholesterol mixed membrane is related to the concentration of AmB. The higher the concentration of AmB, the more likely it is to remove cholesterol from the unsaturated phospholipid membrane. The results are helpful to understand the mechanism of AmB's toxicity to the erythrocyte's membrane, which has a guiding value for seeking ways to reduce the AmB's toxicity.