Concentration- and time-dependence of amphotericin-B induced permeability changes across ergosterol-containing liposomes

Abstract

The effect of amphotericin B on the permeability properties of liposomes prepared by reverse-phase evaporation was examined by using an osmotic method. This study has revealed that the magnitude and type of the alterations in permeability induced by amphotericin B in liposomes made of egg phosphatidylcholine and ergosterol depend not only on the amphotericin B concentration in the external aqueous solution but also on the time elapsed after mixing. Thus, low amphotericin B concentrations (from 0.2 to 1.2 μM) led to, (1) an small increment of the total extent of shrinkage of liposomes suspended in non-electrolytes such as urea or salts like KNO 3, (2) an enhancement of urea and salt permeabilities at the same time scale at which volume changes were measured (ms to s), (3) a maximal blocking by tetraethylammonium of amphotericin B-induced urea permeability and (4) an enhancement of glucose permeability but only after liposomes were incubated with amphotericin B for some minutes before mixing. The high amphotericin B concentration regime (beyond 1.2 μM) led to, (1) a decrease of the total extent of shrinkage of liposomes immediately after rapid mixing of liposomes with urea solutions containing amphotericin B and (2) a 50% reduction of the tetraethylammonium blocking of amphotericin B-induced urea permeability. These results are explained by assuming that amphotericin B may form in ergosterol-containing liposomes two types of active channel differing in internal diameter.