Estudos de membranas modelo e efeitos de terpenos em membranas de leishmania por ressonância paramagnética eletrônica

• Autor(a) principal: Camargos, Heverton Silva de
• Data de Publicação: 2013
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da UFG
• Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/3559
• topic: Componentes espectrais; RPE; Terpenos; Leishmania; Spectral components; EPR; Terpenes; Leishmania; FISICA::FISICA GERAL

Electron paramagnetic resonance (EPR) spectroscopy of spin labels was used to study the main structural accommodations of environment-sensitive probes in the bilayers of saturated phosphatidylcholines with acyl chains lengths ranging from 16 to 22 carbon atoms. The more detailed analysis were made on the spin probe 5-doxyl methyl stearate (5-DMS) whose EPR spectra allowed to identify two distinct spectral components in thermodynamic equilibrium at temperatures below and above the main phase transition. The EPR spectroscopy distinguishes two components associating lower motion with higher polarity (denoted component 1) and higher motion with lower polarity (component 2), which may be assigned to one shallow (more rigid structure) and one deep population of spin probe, respectively. At temperatures until 22◦C only one spectral component can be noted in the spectra whereas at 30◦C the component 1 coexists with an appreciable fraction of component 2. In the liquid-crystalline phase the 5-MSL showed two spectral components for all studied lipids in the entire range of measured temperatures. An accurate analysis of EPR spectra, performed using two fitting programs (NLLS and EPRSIM), allowed us to obtain the thermodynamic profile to these major probe accommodations. Focusing the analysis on two-component EPR spectra, it was studied the influences of cholesterol and a membrane permeation enhancer on the mobility and distribution of spin label on these two main bilayer environments. Parte II Cutaneous leishmaniasis is a neglected tropical disease that infects millions of people worldwide, representing a serious public health problem. The current treatment is based on chemotherapy, using pentavalent antimonials compounds, which cause serious side effects. Electron paramagnetic resonance (EPR) spectroscopy of the spin-label analog of stearic acid (5-DSA ) was used to monitor the effect of the terpenes α-terpineol, 1,8-cineole, III (+)-limonene and nerolidol on the plasma membrane fluidity of Leishmania amazonensis promastigotes. Cytotoxic effects on the parasite were also measured to investigate the relationship between the cytotoxic potential of terpenes and their ability to alter membrane fluidity. All terpenes increased the fluidity of the cellular membrane, without significant differences at higher concentrations. However, the minimum concentration required to cause a change in the membrane was very different between the terpenes and similar to that caused 50% growth inhibition (IC50) showing a correlation between membrane alterations and cytotoxicity. The IC50 values of terpenes analyzed showed the following relationship: nerolidol < (+)-limonene < α-terpineol < 1,8-cineole, with an IC50 of 8 μM for nerolidol and 4700 μM to 1,8- cineole. The EPR spectra of the maleimide derivative spin label (6-MSL) covalently attached to the Leishmania membrane proteins indicated that the terpenes essentially do not alter the dynamics of protein backbone and only increase the mobility of the nitroxide side chain. Cell lysis was not detected at cytotoxic concentrations, as measured by the presence of spin-labeled membrane fragments. Since the terpenes are considered potent skin permeation enhancers with low irritation potential, this work suggests checking the possibility of terpenes applications in the treatment of tegumentary leishmaniasis, where terpenes could perhaps perform a dual action of be an active principle and at the same time facilitate the penetration of other molecules with antileishmanial activity.