Atividade anti-inflamat??ria de novos compostos sint??ticos da classe dos Nitroestirenos

Submitted by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-29T13:03:50Z No. of bitstreams: 1 Disserta????o - M??rcia de Jesus A. Silva.pdf: 2397176 bytes, checksum: 10d2601e2d4c7bb5bd425131e63a0ebc (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-29T13:04:06Z (GMT) No. of bitstreams: 1 Disserta????o - M??rcia de Jesus A. Silva.pdf: 2397176 bytes, checksum: 10d2601e2d4c7bb5bd425131e63a0ebc (MD5) Approved for entry into archive by Divis??o de Documenta????o/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-29T13:04:27Z (GMT) No. of bitstreams: 1 Disserta????o - M??rcia de Jesus A. Silva.pdf: 2397176 bytes, checksum: 10d2601e2d4c7bb5bd425131e63a0ebc (MD5) Made available in DSpace on 2016-11-29T13:04:27Z (GMT). No. of bitstreams: 1 Disserta????o - M??rcia de Jesus A. Silva.pdf: 2397176 bytes, checksum: 10d2601e2d4c7bb5bd425131e63a0ebc (MD5) Previous issue date: 2016-04-29 CAPES - Coordena????o de Aperfei??oamento de Pessoal de N??vel Superior Inflammation is a physiological process that begins in response to infection or tissue damage. It is a mechanism focusing on tissue repair after injury and consists of a cascade of cellular and microvascular events that aims to remove damaged tissue and generation of new ones. In this cascade is observed an increasing in microvascular permeability, followed by adhesion and cell infiltration at the site of injury, apoptosis, and growth of new tissue and blood vessels. The nitrostyrene is a group of compounds consisting of an aromatic ring linked to a chain of the nitro group (NO2). It has been studied by having various biological activities such as anti-apoptotic, anti-platelet or anti-microbial, although few studies have been made for the anti-inflammatory activity of these compounds. The aim of this study was to evaluate anti-inflammatory activity of four new compounds from the class of nitrostyrene (NPA, 7B, 7D and 7E) in in vitro/in vivo assays. Initially, the compounds were submitted to in vitro testing in a murine macrophages model activated by lipopolysaccharide (LPS) and MRC5 human lung fibroblast in order to assess the cytotoxic potential of such compounds. Shortly thereafter, they were evaluated for inhibition of nitric oxide (NO??), for dosage of the cytokine TNF-?? and evaluation of the expression of proteins involved in the inflammatory process. To evaluate the acute inflammatory process in vivo was performed edema induction by carrageenan tests on zebrafish model of LPS-induced pleurisy in mice. The four new compounds tested showed cytotoxic activity on J774A.1 and MRC5 cell when tested in the concentrations (3.12-25 ??g/ml). For THP-1 cytotoxic activity was starting concentration of 10 ??g/ml. All compounds inhibited nitric oxide production by macrophages activated by LPS but only the NPA showed a possibly significant concentration-response at the concentrations of 10 and 20 ??g/ml. This same compound also reduced TNF-??, NF-???B and ERK 1/2 levels in THP-1 cells. The NPA possibly also inhibited the carrageenan-induced edema in Zebrafish fish and leukocyte migration into the pleural cavity of mice by inhibiting the intracellular signaling pathways NF-???B and ERK 1/2 in in vivo assays. In conclusion, compounds tested in this study not only presented significant cytotoxic activity, as also showed an excellent in vitro/in vivo anti-inflammatory potential, most likely by acting in the inhibition of NF-???B and ERK 1/2 pathways. Data were important to explore the biological activity of these novel compounds and their possible application in the treatment of inflammatory diseases. A inflama????o ?? um processo fisiol??gico que se inicia em resposta a uma infec????o ou dano aos tecidos. ?? um mecanismo direcionado ao reparo tecidual ap??s a les??o e consiste em uma cascata de eventos celulares e microvasculares que tem como objetivo a remo????o de tecidos danificados e a gera????o de outros novos. Nessa cascata ?? observado o aumento da permeabilidade microvascular, seguido pela ades??o e infiltra????o de c??lulas no local da les??o, apoptose celular e crescimento de novos tecidos e vasos sangu??neos. Os nitroestirenos s??o um grupo de compostos constitu??do por anel arom??tico ligado a uma cadeia do grupo nitro (NO2). Eles t??m sido estudados por apresentar diversas atividades biol??gicas tais como anti-apopt??tica, antiplaquet??ria e antimicrobiana, entretanto poucas pesquisas t??m sido realizadas quanto ?? atividade anti-inflamat??ria desses compostos. O objetivo deste estudo foi avaliar atividade anti-inflamat??ria de quatro novos compostos da classe dos nitroestirenos (NPA, 7B, 7D e 7E) em ensaios in vitro/in vivo. Inicialmente, os compostos foram submetidos a ensaio in vitro em modelo de macr??fagos murino ativados por lipopolissacar??deo (LPS) e fibroblastos de pulm??o humano MRC5, a fim de avaliar a capacidade citot??xica desses compostos. Logo em seguida, foram avaliados quanto ?? inibi????o de ??xido n??trico (NO??), dosagem da citocina TNF-?? e avalia????o da express??o de prote??nas envolvidas com o processo inflamat??rio. Para avaliar o processo anti-inflamat??rio agudo in vivo foi realizado ensaios de indu????o do edema por carragenina em peixe Zebrafish e o modelo de pleurisia induzido por LPS em camundongos. Os quatro novos compostos testados apresentaram atividade citot??xica em c??lula J774A.1 e MRC5 quando testados nas concentra????es (3.12-25 ??g/mL). Para THP-1 a atividade citot??xica foi apartir da concentra????o 10 ??g/mL. Todos os compostos inibiram a produ????o de ??xido n??trico em macr??fago ativado por LPS, entretanto somente NPA apresentou uma poss??vel concentra????o-resposta significativa nas concentra????es de 10 e 20 ??g/mL. Este mesmo composto tamb??m reduziu os n??veis de TNF-??, NF-???B e ERK 1/2 em c??lulas THP-1. O NPA tamb??m possivelmente inibiu o edema induzido por carragenina em peixe Zebrafish e a migra????o de leuc??citos na cavidade pleural de camundongos atrav??s da inibi????o das vias de sinaliza????o intracelular NF-???B e ERK 1/2 em ensaios in vivo. Em conclus??o, compostos testados neste estudo nao somente apresentaram atividade citot??xica significativa, como tamb??m demonstraram um excelente potencial anti-inflamat??rio in vitro e in vivo, provavelmente por atua????o de inibi????o das vias NF-???B e ERK 1/2. Os dados obtidos foram importantes para explora????o da atividade biol??gica destes novos compostos e sua poss??vel aplica????o na terap??utica de doen??as inflamat??rias.