Sterol Methenyl Transferase Inhibitors Alter the Ultrastructure and Function of the Leishmania amazonensis Mitochondrion Leading to Potent Growth Inhibition

Abstract

We describe here the effects of Delta(24(25)) sterol methenyl transferase inhibitors (SMTI) on promastigote and axenic amastigote forms of Leishmania amazonensis. When these cells were exposed to 20-piperidin-2-yl-5alpha-pregnan-3beta-20-diol (22,26-azasterol; AZA), hydrazone-imidazol-2-yl-5alpha-pregnan-3beta-ol (IMI), 20-hydrazone-pyridin-2-yl-5alpha-pregnan-3beta-ol (PYR) or 24(R,S),25-epiiminolanosterol (EIL), a concentration- and time-dependent inhibition of growth was observed, with IC(50) values in the sub-micromolar range. Ultrastructural alterations in treated cells were mainly observed in the mitochondrion, which displayed an intense swelling and a reduction of the electron density of the matrix with remarkable changes in the inner mitochondrial membranes. Mitochondrial transmembrane electric potential (DeltaPsi) was measured using spectrophotometric methods in control and treated promastigotes permeabilized with digitonin. After energization with the substrates for complexes I, II or IV of the respiratory chain, it was possible to detect marked changes of DeltaPsi in promastigotes treated with 1 microM of the SMTI for 48 or 72 h when compared with normal cells, indicating that these compounds led to the loss of the energy-transducing properties of the mitochondrial inner membrane, probably related to the alteration of its lipid composition. The present study confirms these findings, showing that in Leishmania amazonensis the mitochondrial complex appears to be the first organelle affected after treatment with different SMTI.