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Previous issue date: 2017-03-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Em certas circunst?ncias, Candida albicans pode passar de colonizante para infectante e, como na candid?ase oral. A morfog?nese de C. albicans, bem como sua capacidade de combater o estresse oxidativo no interor de c?lulas fagoc?ticas, ? um fator essencial para a invas?o tecidual e estabelecimento da infec??o. Devido ao baixo arsenal antif?ngico dispon?vel no mercado e o constante surgimento de cepas resistentes, faz-se necess?ria a pesquisa de novas fontes terap?uticas, principalmente oriundas de produtos naturais. No presente estudo foram selecionados 48 isolados cl?nicos de C. albicans oriundos da cavidade bucal de pacientes transplantados renais. A fra??o acetato de etila de Eugenia uniflora foi utilizada na concentra??o de 1000 ?g/mL para avaliar a a??o comparativa (tratada e n?o tratada com extrato) sobre a fagocitose e morfog?nese de C. albicans. Foi realizado o ensaio de resist?ncia ao ataque de neutr?filos polimorfonucleares. O isolado 111R, de alta capacidade filamenta??o foi utilizado para avalia??o do perfil prot?ico por meio de an?lise prote?mica, bem como da intera??o com prote?nas diretamente associadas ? morfog?nese. A resposta ? infec??o foi observada em modelo murino de candid?ase oral e a a??o t?xica do extrato de E. uniflora foi observada em ensaio de MTT. O extrato de E. uniflora reduziu significativamente a capacidade de fagocitose de C. albicans (m?dia total de 120.36 ? 36.71 vs. 44.68 ? 19.84). Trinta e nove prote?nas foram identificadas na an?lise prote?mica, relacionadas ? gera??o de energia, metabolismo de prote?nas e glicose, divis?o celular, transporte citoplasm?tico, metabolismo de ?cidos nucl?icos, estrutura celular e resposta ao estresse. Importantes prote?nas relacionadas com a forma??o do citoesqueleto foram reguladas negativamente nas c?lulas tratadas. Houve instala??o de infec??o na cavidade oral dos camundongos e a infec??o foi atenuada quando C. albicans foi pr?-incubada na presen?a do extrato de E. uniflora e quando o extrato foi aplicado na cavidade oral ap?s a instala??o da infec??o. Este resultado foi condizente com a redu??o na contagem de UFC (2.36 vs. 1.85 Log10 CFU/ml) e a atenua??o dos danos teciduais observados na an?lise histopatol?gica. O extrato de E. uniflora n?o foi t?xico para c?lulas humanas mesmo em concentra??es 8x acima da utilizada nos experimentos. A fra??o acetato de etila de E. uniflora poder? causar danos ? parede celular e prote?nas essenciais ao metabolismo de C. albicans, afetando prote?nas relacionadas ? estrutura celular, reduzindo a capacidade pl?stica de filamenta??o, atenuando a a??o invasiva em modelo animal, sem causar efeito t?xico em c?lulas humanas, podendo ser uma futura alternativa terap?utica para o tratamento de infec??es por Candida. / Under certain circumstances, Candida albicans may switch a colonizing to a infecting yeast, such as in oral candidiasis. Morphogenesis in C. albicans, besides its ability to combat the oxidative stress inside phagocytic cells is an essential step for tissue invasion and establishment of infection. Due to the reduced drug arsenal used for treatment of fungal infections and the constant emergence of resistant strains, it is mandatory to search for new therapeutic sources, mainly from natural products. In the present study, 48 clinical isolates of C. albicans from the oral cavity of renal transplant recipients were selected. In order to evaluate the comparative action (treated and untreated cellst) on phagocytosis and morphogenesis of C. albicans, the ethyl acetate fraction of Eugenia uniflora extract was used at a concentration of 1000 ?g/mL. Resistance of C. albicans to polymorphonuclear neutrophils was carried out. The isolate 111R, a highly filamentous strain was used to evaluate protein profiling through proteomic analysis, as well as the interaction with proteins directly associated with morphogenesis. The in vivo response to infection was observed in murine model of oral candidiasis and the toxic action of E. uniflora extract was observed in the MTT assay. E. uniflora extract significantly reduced the phagocytosis of C. albicans (Mean 120.36 ? 36.71 vs. 44.68 ? 19.84). Thirty-nine proteins, related to energy generation, protein and glucose metabolism, cell division, cytoplasmic transport, nucleic acid metabolism, cell structure and stress response were identified with proteomics analysis. Important proteins directly related with cytoesqueleton formation were down regulated on treated cells. The infection in the oral cavity of the mice was established and it was attenuated when both C. albicans cells were either preincubated in the presence of E. uniflora extract or when the extract was applied to the surface of the oral cavity after infection. These results were consistent with the reduction in CFU countings (2.36 vs. 1.85 Log10 CFU/ml) and attenuation of tissue damages observed in the histopathologycal analysis. E. uniflora extract was non-toxic to human cells even at concentrations 8 fold hugher than the one used in the experiments. The ethyl acetate fraction of E. uniflora may act act damaging and metabolism essential proteinsmainly related to cellular structure, reducing the plastic capacity of filamentation and attenuating infection in a murine modell model, without causing any toxic effect on human cells, suggesting that it may be a future therapeutic alternative for the treatment of Candida infections.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/23748 |
| Date | 30 March 2017 |
| Creators | Silva, Walicyranison Plinio da |
| Contributors | 25849518860, http://lattes.cnpq.br/1463249528959656, Melo, Analy Salles de Azevedo, 01170549845, http://lattes.cnpq.br/8922840487452492, Luchessi, Andr? Ducati, 30489740839, http://lattes.cnpq.br/4420863418928278, Silva, Marcelo de Sousa da, 85074276420, http://lattes.cnpq.br/1295430560645312, Batista, Wagner Luiz, 02674560932, Chaves, Guilherme Maranh?o |
| Publisher | PROGRAMA DE P?S-GRADUA??O EM CI?NCIAS DA SA?DE, UFRN, Brasil |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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