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Previous issue date: 2017-03-03 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The genus Baccharis belongs to the family Asteraceae and has several species widely used in folk medicine and own many compounds of pharmaceutical interest. The aim of this work was to evaluate the cytotoxic, antiproliferative effect and phenotypic reversion of Baccharis anomala extract on activated hepatic stellate cells (HSCs) and identify the compounds present in the extract. In order to evaluate the cytotoxic and antiproliferative effect of the fractionated extract, lactate dehydrogenase (LDH) release and the trypan blue exclusion method were used. The phenotype reversion was evaluated by staining with Oil Red and PPAR-? expression by RT-PCR. The fractions obtained from the methanolic extract were characterized for their phenolic composition by HPLC. The results showed that two fractions of methanolic extract decreased cell proliferation without increasing LDH release levels. Cell cycle arrest was evaluated using 7-AAD and DAPI staining was used to evaluate indications of apoptosis. Fractions of B.anomala induced cell cycle arrest in G1 phase, while DAPI staining did not reveal cell death by apoptosis. Oil-Red (ORO) staining showed the ability of fractions to induce phenotypic reversion and the evaluation of PPAR-? mRNA expression was not altered, suggesting that there is an independent PPAR-? pathway involved. The main components identified in the fractions that presented biological activity were hydroxybenzoic, chlorogenic and coumaric acid. Our study was able to show the antiproliferative effect of methanolic extract fractions of B.anomala, its potential to reverse the activated phenotype of hepatic stellate cells and showed its potential for the treatment of liver fibrosis. / O g?nero Baccharis pertence ? fam?lia Asteraceae e tem v?rias esp?cies amplamente utilizadas na medicina popular, possuindo muitos compostos de interesse farmac?utico. O objetivo deste trabalho foi avaliar o efeito citot?xico, antiproliferativo e a revers?o fenot?pica do extrato de Baccharis anomala em c?lulas estreladas hep?ticas ativadas (HSCs) e identificar os compostos presentes no extrato. Para avaliar o efeito citot?xico e antiproliferativo do extrato fracionado, foram utilizadas a libera??o de lactato desidrogenase (LDH) e o m?todo de exclus?o de azul de Tripan. A revers?o do fen?tipo foi avaliada por colora??o com Oil Red e pela express?o de PPAR-? por RT-PCR. As fra??es obtidas a partir do extrato metan?lico foram caracterizadas pela sua composi??o fen?lica por HPLC. Os resultados mostraram que duas fra??es do extrato metan?lico diminu?ram a prolifera??o celular sem aumentar os n?veis de libera??o de LDH. A parada do ciclo celular foi avaliada usando 7-AAD e colora??o por DAPI foi utilizada para avaliar a apoptose celular. As fra??es de B.anomala induziram a parada do ciclo celular em fase G1, a colora??o por DAPI n?o revelou morte celular por apoptose. Colora??o por Oil-RedO (ORO) mostrou a capacidade de indu??o da revers?o fenot?pica, a avalia??o da express?o mRNA de PPAR-y por RT-PCR n?o foi alterada, sugerindo que exista uma via independente de PPAR-y envolvida. Os principais componentes identificados nas fra??es que apresentaram atividade biol?gica foram os ?cidos hidroxibenz?ico, clorog?nico e cum?rico. Nosso estudo foi capaz de mostrar o efeito antiproliferativo das fra??es de extrato metan?lico de B.anomala, seu potencial para reverter o fen?tipo ativado das c?lulas estreladas hep?ticas e mostrou seu potencial para o tratamento da fibrose hep?tica.
| Identifer | oai:union.ndltd.org:IBICT/oai:tede2.pucrs.br:tede/7615 |
| Date | 03 March 2017 |
| Creators | Basso, Bruno de Souza |
| Contributors | Oliveira, Jarbas Rodrigues, Santar?m, Eliane Romanato |
| Publisher | Pontif?cia Universidade Cat?lica do Rio Grande do Sul, Programa de P?s-Gradua??o em Biologia Celular e Molecular, PUCRS, Brasil, Faculdade de Bioci?ncias |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da PUC_RS, instname:Pontifícia Universidade Católica do Rio Grande do Sul, instacron:PUC_RS |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 8198246930096637360, 500, 500, 500, 600, 36528317262667714, -1634559385931244697, 2075167498588264571 |
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