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Previous issue date: 2015 / A Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob., (Asteraceae) ? nativa da flora brasileira, e sua prepara??o alco?lica tem sido utilizada de forma t?pica como anti-inflamat?rio na medicina popular. Tendo em vista a aus?ncia de estudos acerca da bioatividade e composi??o qu?mica dessa esp?cie, o objetivo do presente estudo foi avaliar o efeito de prepara??es de partes a?reas de P.brasiliensis sobre par?metros inflamat?rios, in vitro, e realizar a an?lise dos seus constituintes qu?micos. A triagem fitoqu?mica dos caules, flores e folhas da planta foi realizada por meio de rea??es cromog?nicas, fluorog?nicas e de precipita??o e sugeriu a presen?a de saponinas, terpenos, taninos e flavonoides nas partes a?reas da planta. Os extratos etan?licos de caules (PBETca), flores (PBETfl) e folhas (PBETfo) foram avaliados por cromatografia l?quida de alta efici?ncia acoplada a detector de arranjo de diodos (CLAE-DAD). Essa an?lise sugeriu a presen?a de flavonoides nos extratos, especialmente das classes das flavonas e flavon?is. Os ?leos essenciais de flores (PBOEfl) e de folhas (PBOEfo) foram analisados por Cromatografia Gasosa acoplada a Espectr?metro de Massas (CG-EM), sendo poss?vel a identifica??o de 22 compostos no PBOEfl e 27 no PBOEfo, dentre os quais, o ?-pineno, limoneno, ?-pineno, sabineno, ?xido de cariofileno e E-cariofileno est?o presentes em maior valor percentual. Foram confeccionadas culturas de 4h de sangue total para avaliar o efeito hemol?tico dos extratos, determinado pela densidade ?ptica do sobrenadante em 540 nm. Houve indu??o de hem?lise em culturas tratadas com PBETca ou PBETfo em concentra??es superiores a 100 ?g/mL. O extrato PBETfl induziu hem?lise em culturas na concentra??o final de 200 ?g/mL. A toxicidade dos produtos naturais ?s C?lulas Mononucleares do Sangue Perif?rico (PBMC) humano ap?s 24 h e 5 dias, foi avaliada por citometria de fluxo, utilizando Azul de Tripan ou Iodeto de Prop?deo. O efeito dos produtos naturais sobre a produ??o de TNF-?, IFN-? e IL-2 foi determinado por ELISA do sobrenadante de culturas de 24 h, ap?s est?mulo com Miristato Acetato de Forbol (PMA) + Ionomicina. Todos os extratos e ?leos essenciais reduziram a produ??o de TNF-?. A s?ntese de IFN-? foi inibida pelo extrato PBETfo e pelo ?leo essencial PBOEfl. Todos os extratos etan?licos e o PBOEfl, inibiram a produ??o de IL-2. O PBETfo foi fracionado por meio de parti??o em solventes com polaridade crescente e todas as fra??es foram eficientes em inibir a s?ntese de TNF-?, por PBMC, ap?s 24 h de cultura. Somente a fra??o hex?nica reduziu a s?ntese de IFN-? e IL-2. Foram confeccionadas culturas de sangue total estimuladas com lipopolissacar?deo (LPS) para investiga??o do efeito dos ?leos essenciais sobre a express?o de
COX-2, por citometria de fluxo. Os ?leos essenciais n?o interferiram na express?o de COX-2. Nossos achados sugerem que o efeito anti-inflamat?rio atribu?do ? P.brasiliensis na medicina popular pode, pelo menos em parte, ser devido ? inibi??o da produ??o de citocinas pr?-inflamat?rias por linf?citos e mon?citos humanos. ? poss?vel que flavonoides das subclasses das flavonas e flavon?is possam contribuir para a atividade biol?gica dos extratos etan?licos de P. brasiliensis. Muitos monoterpenos e sesquiterpenos presentes nos ?leos essenciais de flores e folhas de P. brasiliensis j? tiveram a atividade anti-inflamat?ria descrita em outros trabalhos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / ABSTRACT
The Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob., (Asteraceae) is native from Brazilian flora, and its alcoholic preparation has been used topically as anti-inflammatory in folk medicine. Considering the lack of studies about the bioactivity and chemical composition of this specie, the aim of this study was to evaluate the effect of P. brasiliensis aerial parts preparations on in vitro inflammatory parameters, and perform analysis of its chemical constituents. The plant stems, flowers and leaves phytochemical screening was carried out by chromogenic, fluorogenic and precipitation reactions and suggested the presence of saponins, terpenes, tannins and flavonoids in the aerial parts of the plant. The ethanolic extracts of stems (PBETca), flowers (PBETfl) and leaves (PBETfo) were evaluated by high performance liquid chromatography coupled to diode array detector (HPLC-DAD). These analyses suggested the presence of flavonoids in the extracts, especially the flavones and flavonols subclasses. The flowers (PBOEfl) and leaves (PBOEfo) essential oils were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), and it was possible to identify 22 and 27 compounds in PBOEfl and PBOEfo, respectively. Among the compounds the ?-pinene, limonene, ?-pinene, sabinene, caryophyllene oxide and E-caryophyllene were present in high porcentage. A 4h culture of whole blood was prepared to evaluate the hemolytic effect of the extracts, determined by supernatant optical density at 540 nm. There were hemolysis induction in cultures treated with PBETca or PBETfo at concentrations exceeding 100 ?g/mL. The extract PBETfl induced hemolysis in cultures at a final concentration of 200 ?g/mL. The natural products toxicity to human Peripheral Blood Mononuclear Cells (PBMCs) after 24 h and 5 days were assessed by flow cytometry using trypan blue or propidium iodide. The effect of non-toxic concentrations of the natural products on TNF-?, IFN-? and IL-2 production was determined by ELISA of 24h supernatants culture after stimulation with phorbol myristate acetate (PMA) + Ionomycin. All extracts and essential oils reduced the TNF-? production. The IFN-? synthesis was inhibited by PETfo extract and PBOEfl essential oil.The PBETfo and PBOEfl inhibited the IFN-? synthesis. The PBOEfl and all ethanolic extracts inhibited IL-2 production. The PBETfo was fractionated by solvent partitioning of increasing polarity and all fractions were effective in inhibiting the TNF-? synthesis by PBMC after 24 h culture. Only the hexane fraction reduced the IFN-? and IL-2 synthesis. Whole blood cultures stimulated with lipopolysaccharide (LPS) were prepared to investigate the effect of essential
oils on COX-2 expression by flow cytometry. Essential oils did not affect the expression of COX-2. Our findings suggest that the anti-inflammatory effect attributed to P.brasiliensis in folk medicine can, at least in part, be due to the inhibition of proinflammatory cytokines production by human lymphocytes and monocytes. It is possible that flavonoids of flavones and flavonols subclasses may contribute to the biological activity of ethanolic extracts of P. brasiliensis. Many monoterpenes and sesquiterpenes present in essential oils of P. brasiliensis flowers and leaves already had the anti-inflammatory activity described in other studies.
| Identifer | oai:union.ndltd.org:IBICT/oai:acervo.ufvjm.edu.br/jspui:1/798 |
| Date | 30 January 2015 |
| Creators | Almeida, Val?ria Gomes de |
| Contributors | Melo, Gustavo Eust?quio Brito Alvim de, Rocha-Vieira, Etel, Lopes, Miriam Teresa Paz, Santos, Raquel Gouveia dos, Pereira, Wagner de F?tima, Universidade Fedaral dos Vales do Jequitinhonha e Mucuri (UFVJM), Melo, Gustavo Eust?quio Brito Alvim de |
| Publisher | UFVJM |
| 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 UFVJM, instname:Universidade Federal dos Vales do Jequitinhonha e Mucuri, instacron:UFVJM |
| Rights | A concess?o da licen?a deste item refere-se ao ? termo de autoriza??o impresso assinado pelo autor, assim como na licen?a Creative Commons, com as seguintes condi??es: Na qualidade de titular dos direitos de autor da publica??o, autorizo a Universidade Federal dos Vales do Jequitinhonha e Mucuri e o IBICT a disponibilizar por meio de seus reposit?rios, sem ressarcimento dos direitos autorais, de acordo com a Lei n? 9610/98, o texto integral da obra disponibilizada, conforme permiss?es assinaladas, para fins de leitura, impress?o e/ou download, a t?tulo de divulga??o da produ??o cient?fica brasileira, e preserva??o, a partir desta data., info:eu-repo/semantics/openAccess |
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