Regulation of calcium ions on the interaction between amphotericin B and cholesterol-rich phospholipid monolayer in LE phase and LC phase

Abstract

Amphotericin B, as a "gold standard", is used to treat invasive fungal infections. The AmB molecule can bind easily to cholesterol and damage cell membranes, so it produces the toxicity on cell membrane, which limits its clinical dose. However, the interaction between AmB and cholesterol-rich membrane is unclear now. The phase state of the membrane and the metal cation outside cell membrane may affect the interaction between AmB and the membrane. In this work, the effects of amphotericin B on the mean molecular area, elastic modulus and stability of mammalian cell membrane rich in cholesterol in the presence of Ca²⁺ ions were studied using DPPC/Chol mixed Langmuir monolayer as a model. The Langmuir-Blodgett method and AFM test were used to study the effects of this drug on the morphology and height of cholesterol-rich phospholipid membrane in the presence of Ca²⁺ ions. The influence of calcium ions on the mean molecular area and the limiting molecular area was similar in LE phase and in LC phase. The calcium ions made the monolayer more condensed. However, calcium ions can weaken the shortening effect of AmB on the relaxation time of the DPPC/Chol mixed monolayer in LE phase but enhance it in LC phase. Interestingly, calcium ions caused a LE-LC coexistence phase to occur in the DPPC/Chol/AmB mixed monolayers at 35mN/m, which was confirmed by atomic force microscopy. The results can help to understand the interaction between amphotericin B and cell membrane rich in cholesterol in the calcium ions environment.

Keywords: Amphotericin B; Atomic force microscopy; Calcium ions; Langmuir monolayer; Relaxation time.