Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T15:38:43Z No. of bitstreams: 2 bethania_alves_avelar_freitas.pdf: 67938565 bytes, checksum: 4f06416afedd4040fb61d7bad494c673 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T15:40:36Z (GMT) No. of bitstreams: 2 bethania_alves_avelar_freitas.pdf: 67938565 bytes, checksum: 4f06416afedd4040fb61d7bad494c673 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T15:41:09Z (GMT) No. of bitstreams: 2 bethania_alves_avelar_freitas.pdf: 67938565 bytes, checksum: 4f06416afedd4040fb61d7bad494c673 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2014-12-17T15:41:09Z (GMT). No. of bitstreams: 2 bethania_alves_avelar_freitas.pdf: 67938565 bytes, checksum: 4f06416afedd4040fb61d7bad494c673 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2013 / Ageratum fastigiatum ? uma planta utilizada na medicina popular como anti-inflamat?rio e analg?sico, no entanto, poucos estudos foram realizados a fim de detalhar os mecanismos envolvidos nessa atividade. O objetivo deste estudo foi avaliar a atividade anti-inflamat?ria in vitro do ?leo essencial e do extrato dicloromet?nico de A. fastigiatum. Pela t?cnica de exclus?o do azul de tripam por citometria de fluxo foram determinadas concentra??es n?o t?xicas das prepara??es de A. fastigiatum. As concentra??es n?o t?xicas do ?leo essencial foram 5x10-3 e 1x10-2 ?L/mL. Essas concentra??es foram utilizadas para a pesquisa do potencial anti-inflamat?rio do ?leo essencial, medido por meio da an?lise do perfil de citocinas pro (TNF- ? e IFN- ?) e anti-inflamat?rias (IL-10), em culturas de leuc?citos humanos estimulados e n?o estimulados com PMA (acetato de forbol miristato) . Os dados demonstraram que ambas as concentra??es inibiram o percentual de linf?citos-TNF+ nas culturas estimuladas com PMA. A an?lise cromatogr?fica em fase gasosa acoplada a espectrometria de massas (CG/EM) revelou como principais constituintes no ?leo essencial as subst?ncias ?-pineno (7,51%), limoneno (5,9%), ?xido de cariofileno (13,59%), 1,2 ep?xido humuleno (8,41%) e 1,6-humulanodien-3-ol (17,71%). O extrato dicloromet?nico de A. fastigiatum, na concentra??o 20 ?g/mL, n?o apresentou toxicidade aos leuc?citos do sangue perif?rico humano e reduziu o percentual de linf?citos-TNF-?+ e linf?citos-IFN-?+ nas culturas estimuladas com o PMA. Este extrato foi fracionado em coluna de Sephadex LH-20 (150 g). Na an?lise de citocinas, a fra??o 10 (AFDM 10), na concentra??o 10 ?g/mL, demonstrou efeito anti-inflamat?rio in vitro reduzindo a frequ?ncia de linf?citos-TNF-?+ e a prolifera??o de linf?citos estimulados com PHA (fitohemaglutinina). Sugere-se que parte da atividade anti-inflamat?ria de A. fastigiatum se d? pela inibi??o que os constituintes da planta promovem sobre a ativa??o de leuc?citos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2013. / ABSTRACT Ageratum fastigiatum is a plant used in folk medicine as topical anti-inflammatory and analgesic agent. However, few studies have been conducted with this plant in order to detail the molecular mechanisms involved in its biological activities. Therefore, main purpose of the present study was to investigate the anti-inflammatory activity of the essential oil and leaf rinse dichloromethane of A. fastigiatum extract. The cytotoxicity of the compounds was assessed by means of trypan blue exclusion. The essential oil at 5x10-3 e 1x10-2 ?L/ did not present cytotoxic activity. These two concentrations were used for investigating the anti-inflammatory potential of the essential oil, by analyzing the pro-inflammatory (TNF-? and IFN-?) and anti-inflammatory (IL-10) cytokine profiles on human leukocyte cultures stimulated and non-stimulated with PMA (phorbol myristate acetate). The data obtained by flow cytometry demonstrated that both concentrations decreased the percentage of lymphocytes ?TNF-? + on the cell cultures stimulated with PMA. In the non-stimulated cell cultures there were no changes in the percentage of IL-10+ cells. The gas phase chromatography coupled with mass spectrometry analysis (GC/MS) revealed that the main compounds present in oil samples were: ?-pinene (7.51%), limonene (5.9%), caryophyllene oxide (13.59%), 1,2 humulene epoxide (8.41%) and 1,6-humulanodien-3-ol (17.71%). The dichloromethane extract of A. fastigiatum, obtained from the washing leaves, at the concentration 20 ?g/mL, did not presented cytotoxicic action on the human peripheral blood leukocytes. Following the same protocol performed to cytokines analysis used before, we observed that the dichloromethane A. fastigiatum extract also decreased the percentage of lymphocytes-TNF-? + and IFN-?+ on the cell cultures stimulated with PMA. This extract after fractionated on Sephadex LH-20 . The dichloromethane A. fastigiatum fraction (AFDM 10) at 10 ?g/mL reducing the frequency of lymphocytes-TNF-?+. Furthermore AFDM 10 decreasing the proliferation of lymphocytes stimulated with PHA (phytohemagglutinin). It is concluded that part of the anti-inflammatory activity of A. fastigiatum is give by inhibiting the plant constituents on promoting the activation of leukocytes.