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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

CaracterizaÃÃo estrutural preliminar e efeitos na inflamaÃÃo da lectina da alga marinha verde Caulerpa cupressoides / Structural characterization and preliminary effects on inflammation of the lectin of the green seaweed Caulerpa cupressoides

Ismael Nilo Lino de Queiroz 19 February 2013 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / As lectinas de algas marinhas possuem vÃrias aplicaÃÃes farmacolÃgicas. O objetivo deste trabalho foi caracterizar parcialmente a estrutura e avaliar os efeitos em modelos clÃssicos de inflamaÃÃo da lectina da alga marinha verde Caulerpa cupressoides. A lectina de Caulerpa cupressoides (LCc) foi extraÃda com tampÃo Tris-HCl 25 mM, pH 7,5 e isolada por cromatografia de troca iÃnica em coluna de DEAE-celulose. A caracterizaÃÃo estrutural parcial apresentou uma sequÃncia aminoterminal com 31 resÃduos de aminoÃcidos, obtida de acordo com o mÃtodo de degradaÃÃo de Edman, enquanto que na espectroscopia de ressonÃncia magnÃtica nuclear unidimensional 1H para a LCc purificada por Sephadex G-100 e obtida por DEAE-celulose, foi demostrado uma similaridade entre os sinais obtidos em ambos os espectros. A atividade anti-inflamatÃria foi avaliada em ratos Wistar machos (n=6), utilizando o modelo de edema de pata induzidos por carragenana (700 μg/pata), dextrana (500 μg/pata), histamina (100 μg/pata), serotonina (20 μg/pata) ou bradicinina (30 μg/pata). Grupos de animais foram submetidos ao tratamento com LCc (0,1; 1,0 ou 10,0 mg/kg; i.v.), 30 min antes do estÃmulo inflamatÃrio. Foi avaliado o envolvimento da LCc (1,0 mg/kg) na via da Heme oxigenase-1. Foram utilizados grupos que receberam tratamento com LCc ligada ao inibidor mucina (8,0 mg/kg; i.v.) ou somente mucina (8,0 mg/kg; i.v.) e dexametasona (1,0 mg/kg; s.c.) foram utilizados como controles. O efeito edematogÃnico de LCc foi avaliado aplicando as doses de 0,1; 1,0 ou 10 mg/kg (i.pl.). No ensaio de edema de pata induzido por carragenana, LCc reduziu a formaÃÃo de edema sendo confirmado pela determinaÃÃo dos nÃveis teciduais de mieloperoxidase. A LCc (1,0 mg/kg) ligada a mucina nÃo apresentou efeito anti-inflamatÃrio no edema de pata induzido por carragenana, exceto na primeira hora apÃs o estÃmulo. No edema induzido por dextrana, LCc tambÃm inibiu o edema osmÃtico. Apenas a dose de 1,0 mg/kg foi utilizada no edema induzido por histamina reduzindo em 40% o edema no intervalo de 30 min LCc (1,0 mg/kg), entretanto, nÃo reduziu a formaÃÃo de edema induzido por serotonina ou bradicinina. AlÃm disso, na anÃlise histolÃgica do tecido subplantar, LCc (1,0 e 10,0 mg/kg) foi capaz de reduzir a migraÃÃo celular. Na presenÃa de ZnPP IX (3,0 mg/kg; s.c.), LCc perdeu sua capacidade inibir o edema induzido por carragenana, exercendo seu mecanismo de aÃÃo anti-inflamatÃrio atravÃs do envolvimento da via da HO-1. Enquanto que na imunohistoquÃmica, LCc (10 mg/kg) reduziu a expressÃo de IL-1β, porÃm ocorreu intensa marcaÃÃo das citocinas TNF-α e IL-6, alÃm da expressÃo de HO-1, nos grupos tratados com a mesma dose de LCc. Com relaÃÃo ao efeito edematogÃnico, LCc foi capaz de induzir intenso processo inflamatÃrio com efeito dose-dependente. No entanto, o edema induzido com a dose de 10 mg/kg de LCc foi foi inibido por indometacina, meclizina, pentoxifilina e dexametasona. Portanto, a LCc quando parcialmente caracterizada apresentou na sua sequÃncia aminoterminal uma identidade de 43% com a proteÃna da alga verde unicelular Chlamydomonas reinhardtii e apresentou propriedades anti- e prÃ-inflamatÃrias, sendo considerada um agente terapÃutico em potencial para estudos futuros nos processos inflamatÃrios. / Lectins seaweed have various pharmacological applications. This work was partially characterize the structure and evaluate the effects on classical models of inflammation lectin of the green seaweed Caulerpa cupressoides. The Caulerpa cupressoides lectin (CcL) was extracted with Tris-HCl 25 mM, pH 7.5 and isolated by ion exchange chromatography on DEAE-cellulose. Structural characterization showed a partial aminoterminal sequence with 31 amino acid residues, obtained according to the method of Edman degradation, while in nuclear magnetic resonance spectroscopy to the CcL 1H-NMR purified by Sephadex G-100 and DEAE-cellulose obtained by was demonstrated similarity between signals obtained in both spectra. The anti-inflammatory activity was evaluated in male Wistar rats (n = 6), using the model of paw edema induced by carrageenan (700 μg/paw), dextran (500 μg/paw), histamine (100 μg/paw), serotonin (20 μg/paw) or bradykinin (30 μg/paw). Groups of animals were treated with CcL (0.1, 1.0 or 10.0 mg/kg, i.v.) 30 min before the inflammatory stimulus. Was evaluated the involvement of CcL (1.0 mg/kg) towards heme oxygenase-1. Groups were used that were treated with inhibitor CcL linked mucin (8.0 mg/kg, i.v.) or only mucin (8.0 mg/kg, i.v.) and dexamethasone (1.0 mg/kg, s.c.) were used as controls. The effect edematogenic CcL was evaluated by applying the doses of 0.1, 1.0 or 10 mg/kg (i.pl.). In the trial of paw edema induced by carrageenan, CcL reduced edema formation was confirmed by determining tissue levels of myeloperoxidase. CcL (1.0 mg/kg) showed no mucin linked to anti-inflammatory effect on the paw edema induced by carrageenan, except in the first hour after stimulation. In dextran-induced edema, CcL also inhibited the osmotic swelling. Only the dose of 1.0 mg/kg was used in reducing histamine-induced edema by 40% the edema within 30 min CcL (1.0 mg/kg), however, did not reduce the edema induced by serotonin or bradykinin. Besides, in the histological analysis of tissue subplantar, CcL (1.0 and 10.0 mg/kg) was able to reduce cell migration. In the presence of ZnPP IX (3.0 mg/kg, s.c.), CcL has lost its ability to inhibit the carrageenan-induced edema, exerting its mechanism of action anti-inflammatory pathway through the involvement of HO-1. While in immunohistochemistry, CcL (10 mg/kg) reduced the expression of IL-1β, but intense staining occurred cytokines TNF-α and IL-6, and expression of HO-1 in the groups treated with the same dose of CcL. In relation the effect edematogenic, CcL was able to induce intense inflammatory process with dose-dependent effect. However, the induced edema at a dose of 10 mg/kg was CcL was inhibited by indomethacin, meclizine, pentoxifylline and dexamethasone. Therefore, when the CcL partially characterized presented in its aminoterminal sequence of 43% identity with the protein from unicellular green alga Chlamydomonas reinhardtii and showed anti-and pro-inflammatory therapeutic agent is considered a potential for future studies in inflammatory processes.

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