Made available in DSpace on 2014-12-17T14:10:28Z (GMT). No. of bitstreams: 1 HylarinaMDS_DISSERT.pdf: 1971584 bytes, checksum: 9428714bda1598d765036d4f0b211af1 (MD5) Previous issue date: 2013-03-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Ulcerative colitis comprising an inflammatory bowel disease, whose most severe consequence is the development of intestinal neoplasia. The drugs currently used to treat the disease trigger a variety of serious adverse effects and are not effective in many cases. Recent studies demonstrated the effectiveness of natural products for the treatment of inflammatory processes. Seaweed extracts and their purified products have shown protective effects in models of inflammation and the association of traditional therapies with probiotics has significantly improved the clinical symptoms of ulcerative colitis. Therefore, the aims of this study include evaluating the potential effects of the use of probiotic strain Enterococcus faecium 32 (Ef32), the methanolic extract of the green seaweed Caulerpa mexicana (M.E.) and their concomitant administration in a murine model of colitis induced by dextran sodium sulfate (DSS). Accordingly, C57BL /6 mice were pretreated orally with Ef32 (109 CFU/ml) for seven days. In the seven days following, the colitis was induced by administration of 3% DSS (w/v) diluted in the animals drinking water. During this period, animals were treated daily with Ef32 and the M.E. (2.0 mg/kg) every other day by intravenous route. The development of colitis was monitored by the disease activity index (DAI), which takes into account the loss of body weight, consistency and presence of blood in stools. After euthanasia, the colon was removed, its length measured and tissue samples were destined for histological analysis and culture for cytokine quantification. The levels of cytokines in the culture supernatant of the colon were measured by ELISA. The treatments with the probiotic Ef32 or the M.E. alone or the combination of these two substances provoked significant improvement as to weight loss and DAI, and prevented the shortening of the colon in response to DSS. The isolated treatments triggered a slight improvement in intestinal mucosal tissue damage. However, their combination was able to completely repair the injury triggered by DSS. The association was also able to reduce the levels of all the cytokines analyzed (IFN-γ, IL-4, IL-6, IL-12, IL-17A and TNF-α). On the other hand, the treatment with Ef32 did not interfere with the levels of TNF-α, whereas treatment with M.E. did not alter the levels of IL-6. Moreover, the treatment with Ef32 not interferes in TNF-α levels, whereas treatment with M.E. did not alter the levels of IL-6. Therefore, the potential probiotic Ef32 and M.E. and especially when these samples were associated proved promising alternatives in the treatment of ulcerative colitis as demonstrated in an experimental model because of its beneficial effects on morphological and clinical parameters, and by reducing the production of proinflammatory cytokines of Th1, Th2 and Th17 / A colite ulcerativa compreende uma doen?a inflamat?ria intestinal, cuja consequ?ncia mais severa ? o desenvolvimento de neoplasia intestinal. Os medicamentos atualmente utilizados para o tratamento da doen?a desencadeiam uma variedade de efeitos adversos s?rios e n?o s?o eficazes em muitos casos. Estudos recentes apontaram para a efic?cia de produtos naturais no tratamento de processos inflamat?rios. Extratos de algas marinhas e seus produtos purificados t?m mostrado efeito protetor em modelos de inflama??o e a associa??o de terapias tradicionais com probi?ticos tem melhorado significativamente o quadro cl?nico da colite ulcerativa. Sendo assim, os objetivos do presente estudo inclu?ram avaliar os efeitos do uso da potencial cepa probi?tica Enterococcus faecium 32 (Ef32), do extrato metan?lico da alga marinha verde Caulerpa mexicana (E.M.) e da administra??o conjunta no modelo murino de colite induzida por dextrana sulfato de s?dio (DSS). Nesse sentido, camundongos C57BL/6 machos foram pr?-tratados oralmente com Ef32 (109 UFC/ml) durante sete dias. Nos sete dias seguintes, a colite experimental foi induzida atrav?s da administra??o DSS 3% (p/v) dilu?da na ?gua de beber dos animais. Neste per?odo, os animais foram tratados com Ef32 diariamente e com o E.M. (2,0 mg/kg) em dias alternados pela rota intravenosa. O desenvolvimento de colite foi monitorado atrav?s do ?ndice de atividade da doen?a (IAD), que leva em considera??o a perda de peso corporal, consist?ncia e presen?a de sangue nas fezes. Ap?s eutan?sia dos animais, o c?lon foi removido e mensurado e amostras do tecido foram destinadas ? an?lise histol?gica e cultura para dosagem de citocinas. Os n?veis de citocinas no sobrenadante da cultura do c?lon foram mensurados por ELISA. Os tratamentos com o probi?tico Ef32 ou com o E.M. sozinhos ou com a associa??o dessas duas subst?ncias desencadearam significativa melhora quanto ? perda de peso corporal e IAD, e preveniram o encurtamento do c?lon em resposta ? DSS. Os tratamentos isolados desencadearam uma melhora discreta no dano tecidual da mucosa intestinal. No entanto, a combina??o deles foi capaz de reparar completamente a inj?ria desencadeada pelo DSS. A associa??o foi ainda capaz de reduzir os n?veis de todas as citocinas analisadas (IFN-γ, IL-4, IL-6, IL-12, IL-17A e TNF-α). Por outro lado, o tratamento com Ef32 n?o interferiu nos n?veis de TNF-α, enquanto que o tratamento com o E.M n?o alterou os n?veis de IL-6. Portanto, o potencial probi?tico Ef32 e o E.M. e especialmente quando essas subst?ncias foram associadas, mostraram-se alternativas promissoras no tratamento da colite ulcerativa, como demonstrado no modelo experimental em raz?o dos seus efeitos ben?ficos sobre par?metros cl?nicos e morfol?gicos, bem como pela redu??o da produ??o de citocinas pr?-inflamat?rias das respostas Th1, Th2 e Th17

Made available in DSpace on 2014-12-17T14:16:27Z (GMT). No. of bitstreams: 1 MarianaAOB_DISSERT.pdf: 3079508 bytes, checksum: 5d34c7cd249531ceb74afa12d1184e1f (MD5) Previous issue date: 2011-02-28 / The regulation of the inflammatory response is essential to maintain homeostasis. Several studies have been performed to search new drugs that can contribute to avoiding or minimizing an excessive inflammatory process. The aim of this study was to evaluate the effect of extracts of green algae Caulerpa mexican in models of inflammation. In mice, the model of peritonitis induced inflammatory zymosan pretreatment of mice with aqueous and methanol extracts of C. mexican was able to reduce cell migration to the peritoneal cavity. Treatment of mice with extracts of C. mexican also reduced the ear edema induced by xylene and exerted inhibitory action on the migration of leukocytes in inflammation-induced zymosan the air pouch, and timedependent for the extracts tested in the model of ulcerative colitis induced by DSS 3%, the extract methanol, but not the aqueous C. mexican, significantly reduced the clinical symptoms of colitis, as well as the production of proinflammatory cytokines in the culture of mouse colon, in the histological analysis there was a slight reduction of inflammation in the intestinal mucosa. We concluded that the administration of the extracts resulted in the reduction of cell migration to different sites as well as reducing the edema formation induced by chemical irritant. This study demonstrates for the first time the antiinflammatory effect of aqueous and methanolic extracts from green marine algae Caulerpa mexican / A regula??o da resposta inflamat?ria ? essencial para manter a homeostase. Diversos estudos t?m sido realizados para a pesquisa de novas drogas que possam contribuir para evitar ou minimizar um processo inflamat?rio excessivo. O objetivo deste estudo foi avaliar o efeito de extratos de algas verde Caulerpa mexicana em modelos de inflama??o. Em camundongos, o modelo de peritonite inflamat?ria induzida por zimosam o tratamento pr?vio de camundongos com extrato aquoso e metan?lico de C. mexicana foi capaz de reduzir a migra??o celular para a cavidade peritoneal. O tratamento dos camundongos com os extratos da C. mexicana tamb?m diminuiu o edema de orelha induzido por xilol e exerceu a??o inibit?ria da migra??o de leuc?citos em inflama??o induzido pelo zimosam na bolsa de ar, sendo tempo-dependente para os extratos testados, no modelo de colite ulcerativa induzida por DSS 3%, o extrato metan?lico, mas n?o o aquoso da C. mexicana, reduziu de forma significativa o quadro cl?nico da colite, como tamb?m a produ??o de citocinas pr?-inflamat?rias na cultura do c?lon do camundongo, na an?lise histol?gica houve uma leve redu??o da inflama??o na mucosa intestinal. Conclu?mos que a administra??o dos extratos resultou na redu??o da migra??o de c?lulas para diferentes modelos experimentais, bem como reduzir a forma??o de edema induzida por irritante qu?mico. Este estudo demonstra pela primeira vez o efeito anti-inflamat?rio do extrato aquoso e metan?lico da alga verde marinha Caulerpa mexicana