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Detec??o in vitro de citocinas intracitoplasm?ticas (interferon gama, fator de necrose tumoral, interleucina 4 e interleucina 10) em leuc?citos humanos tratados com extrato bruto dilu?do de euphorbia tirucalliAvelar, Beth?nia Alves de 13 August 2010 (has links)
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Previous issue date: 2010 / O uso de plantas da fam?lia Euphorbiaceae, principalmente do g?nero Euphorbia, tem sido popularmente difundido para o tratamento de uma variedade de doen?as de natureza infecciosa, tumoral e inflamat?ria. Entre as esp?cies desse g?nero, a Euphorbia tirucalli ? uma planta de uso difundido no Brasil e em algumas regi?es, tal como a do Vale do Jequitinhonha. H? ind?cios de que o l?tex de E. tirucalli tenha atividade antitumoral e antiviral, por?m, pouco se sabe sobre os mecanismos envolvidos nessa a??o. ? prov?vel que o mecanismo de a??o para tais atividades envolva a ativa??o de leuc?citos e a produ??o de citocinas para o direcionamento de uma resposta antiviral e antitumoral efetivas. Diante disso, o presente trabalho avaliou a produ??o de citocinas TNF-?, IFN-?, IL-4 e IL-10 nas popula??es e subpopula??es leuc?cit?rias do sangue perif?rico estimulados com o l?tex bruto da referida planta. Para tal, leuc?citos do sangue perif?rico de vinte indiv?duos h?gidos foram incubados por 4 horas com o l?tex bruto de E. tirucalli dilu?do em Dimetilsulfoxido (DMSO) e constituiu o grupo l?tex (Lt). Culturas celulares incubadas com salina e DMSO, utilizando-se o mesmo per?odo de incuba??o, constitu?ram as culturas controle n?o estimuladas (CC) e controle de solvente (DMSO), respectivamente. Ap?s o per?odo de incuba??o, as c?lulas foram marcadas com anticorpos monoclonais espec?ficos para receptores de superf?cie celular CD4, CD8 e CD14, bem como para as citocinas intracitoplasm?ticas TNF-?, IFN-?, IL-4 e IL-10. A aquisi??o e an?lise dos dados foram realizadas por citometria de fluxo. Os resultados demonstraram um aumento significativo (p < 0,05) no percentual de linf?citos T e sua subpopula??o T CD4 positivos para as citocinas do tipo 1, TNF-? e IFN-?. Essas citocinas s?o respons?veis por ativar o sistema imune para respostas imunoprotetoras frente ?s infec??es virais e processos tumorais. J? os neutr?filos e linf?citos T CD8, nas culturas estimuladas com o l?tex da planta, apresentaram um perfil misto de produ??o de citocinas do tipo 1 e 2. Esses resultados evidenciaram uma resposta do tipo 1 predominante, in vitro, e uma prov?vel ativa??o de mecanismos moduladores, mediados por linf?citos T CD8 e neutr?filos. Este estudo sugere que a utiliza??o popular da planta em infec??es virais e processos tumorais pode ter algum fundamento, tendo em vista a produ??o preferencial de citocinas do tipo 1 em c?lulas estrat?gicas da resposta imune celular, fundamental no combate ?s patologias mencionadas. No entanto, os estudos in vitro aqui realizados n?o s?o suficientes para garantir que a utiliza??o da planta no contexto in vivo possa, de fato, apresentar tais propriedades. Assim, estudos adicionais, incluindo outros ensaios in vitro, bem como ensaios pr?-cl?nicos, s?o fundamentais para o avan?o das pesquisas envolvendo a utiliza??o de E. tirucalli para fins terap?uticos. / ABSTRACT
The use of plants in the family Euphorbiaceae, especially of the genus Euphorbia, has been popularly circulated for the treatment of a variety of infectious, tumoral and inflammatory illnesses. Among the species of this genus, Euphorbia tirucalli is a the plant with widespread use in Brasil and in some regions, such as the Vale do Jequitinhonha. There is evidence that the latex of E. tirucalli has antiviral and antitumor activity, but little is known about the mechanisms involved in this action. It is likely that the mechanism of action for such activities involves leukocyte activation and cytokine production for directing an effective antitumor and antiviral response. Thus, the present study evaluated the production of cytokines TNF-?, IFN-?, IL-4 and IL-10 in the populations and subpopulations in peripheral blood leukocytes stimulated with crude latex of this plant. For such, peripheral blood leukocytes of twenty healthy subjects were incubated for 4 hours with crude latex of E. tirucalli diluted in dimethyl sulfoxide (DMSO) and was the latex group (Lt). Cell cultures incubated with saline and DMSO, using the same incubation period, represent the control cultures that were not stimulated (CC) and solvent control (DMSO), respectively. After the incubation period, cells were stained with monoclonal antibodies specific to cell surface receptors CD4, CD8 and CD14, as well as for TNF-? intracytoplasmic cytokines, IFN-?, IL-4 and IL-10. The acquisition and analysis were performed by flow cytometry. The results showed a significant increase (p <0.05) in the percentage of T lymphocytes and their subpopulation T CD4 positive for type 1 cytokines, TNF-? and IFN-?. These cytokines are responsible for activating the immune system to immunoprotective responses against viral infections and tumor processes. However the neutrophils and T CD8 lymphocytes, in cultures stimulated with the latex of the plant, showed a mixed profile of cytokine production of type 1 and 2.These results showed a response of the type 1 predominant, in vitro, and a probable activation of modulatory mechanisms mediated by T CD8 lymphocytes and neutrophils.This study suggests that the popular use of plant in viral infections and tumor processes may have some foundation, in order to preferential production of type 1cytokines in strategic cells of the cellular immune response, that is essential in combating the diseases mentioned. However, in vitro studies conducted here are not sufficient to ensure that the use of the plant in vivo context may, in fact, possesses such properties. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2010.
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