Estudo do metabolismo energético de Leishmania spp utilizando cromatografia líquida de alta eficiência e ressonância magnética nuclear
Data
2011-09-16
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Universidade Federal de Goiás
Resumo
Leishmaniasis is a disease of the endothelial reticulum system and is caused by protozoans form the Leishmania genus. The promastigote forms of Leishmania sp degradate carbohydrates through the glycolityc pathway and its first reaction occurs inside the glicosome. As they use as energy source glucose, aminoacids and lipids from the culture media the excreted/secreted (E/S) products may be useful as indicative of the energetic and metabolis profile of the parasite and of its biological disturbances. The biochemical analysis of Leishmania metabolism is important to upgrade the parasitological studies with techniques which are adequate to the evaluation of the biochemical profile of the host-parasite relationship towards a better understanding of how these organisms are able to survive in such diverse environments such as the digestory tract of the insect or inside vertebrate hosts tissues. The aim of this study was to perform a study of the energetic metabolism of species and isolates of Leishmania spp. Therefore the in vitro analysis of E/S metabolic products from the parasite within the culture media during logarithmic and stationary growth phases was performed through chromatographic (HPLC) methods. The spectrophometry was used to glucose measurement and the nuclear magnetic resonance (NMR) was implanted aiming the study of the parasite‟s metabolism. Through HPLC it was possible to verify a significant difference in the E/S of organic acids from the carbohydrates metabolism (pyruvate and lactate), intermediary metabolism (citrate, α-ketoglutarate, succinate, fumarate, malate and oxaloacetate) and from the fatty acids oxidation (β-hydroxibutyrate) between differents species and isolates of Leishmania spp. Through the enzymatic spectrophotometric analysis we verified the different concentrations of glucose. The NMR through 13C and 1H detected the following metabolites in pellet and supernatant: alanine, lactate, acetate, oxalate, pyruvate, succinate, glicerol, CO2, ureia and aminoacids (glutamine, valine and arginine). Therefore, through the several methods it was possible to observe that there is difference of E/S metabolites between the isolates and strains of Leishmania spp that utilize several biochemical pathways to energy production and the identification of these pathways may be used to identify each isolate and also establish metabolic differences in the host-parasite relationship.
Descrição
Palavras-chave
Leishmania spp , Bioquímica parasitária , Metabolismo energético , Ácidos orgânicos , CLAE, RMN , Leishmania spp , Parasite biochemistry , Energetic metabolism , Organic acids , HPLC , NMR , Bioquímic parasitária Leishmania spp.; Metabolism energético parasitário ácid orgânicos; Leishmania (Viannia) braziliensis CLAE/RMN
Citação
COSTA, Tatiane Luiza da. Study of energy metabolism of Leishmania spp using high performance liquid chromatography and nuclear magnetic resonance. 2011. 107 f. Tese (Doutorado em Ciências da Saúde) - Universidade Federal de Goiás, Goiânia, 2011.