Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-12T21:06:34Z No. of bitstreams: 1 DenilsonVieira.pdf: 1857476 bytes, checksum: 07173b7d3884285218ba141bb85ab291 (MD5) / Made available in DSpace on 2017-05-12T21:06:34Z (GMT). No. of bitstreams: 1 DenilsonVieira.pdf: 1857476 bytes, checksum: 07173b7d3884285218ba141bb85ab291 (MD5) Previous issue date: 2012-08-31 / Himatanthus drasticus (Mart.) Plumel is popularly known in the North and Northeast Regions of Brazil as janaúba, janaguba, tiborna, jasmim-manga and pau-de-leite, as well. Brazilian traditional medicine knowledge attributes to the species antiinflammatory, analgesic, antitumoral and dewormer properties. Thus, the present study sought to investigate the anti-inflammatory, analgesic and wound-healing effects of the hidroalcoholic extract (HAE) made from Himatanthus dratiscus stem bark, which were collected in São Bento (MA) Brazil. Oral administration of HAE (0.5, 0.75 and 1.0 g/kg) reduced in a dose-dependent manner both oedema’s formation and leukocyte migration in mice submitted to intraplantar injection of 1% carragenin. Similarly, HAE also reduced the number of contortions in the acetic acid-induced writhing test. However, HAE had no effect on pain’s central perception. Corroborating these data, HAE showed significant wound-healing effect since it accelerated the wound contraction time in the mice. Finally, assessment of in vitro antioxidant activity of HAE suggested that phenolic compounds found in the extract is able to scavenger DPPH radical as fast as well-known antioxidants like -tocopherol. Therefore, this study shows that HAE promotes relevant analgesic, anti-inflammatory, woundhealing and antioxidant activity. Moreover, this study contributes to validation of popular use of the species as well as open up new perspectives for future investigations. / Himatanthus drasticus (Mart.) Plumel é popularmente conhecida nas regiões Norte e Nordeste do Brasil como janaúba, janaguba, tiborna, jasmim-manga, pau-de-leite entre outras denominações. Na prática popular, esta espécie é usada como antiinflamatória, antitumoral, analgésica e vermífuga. O presente estudo investigou os efeitos analgésico, anti-inflamatório, cicatrizante e antioxidante do extrato hidroalcoólico das cascas do caule da Himatanthus drasticus (EHA) coletadas no município de São Bento - MA. Quando administrado nas doses de 0,5; 0,75 e 1,0 g/kg (v.o.), o EHA reduziu de forma dependente de dose a formação de edema e a migração leucocitária em camundongos machos, submetidos a injeção intraplantar de carragenina 1%. De forma similar, o EHA reduziu o número de contorções abdominais induzidas por ácido acético 1%, evidenciando a atividade antinociceptiva. Entretanto, o EHA não promoveu qualquer efeito sobre a percepção central da dor. Em consonância com estes efeitos, o EHA também mostrou significante atividade cicatrizante, uma vez que, acelerou o tempo de contração da ferida induzida por excisão de pele nos camundongos. Finalmente, a avaliação da atividade antioxidante in vitro do EHA foi sugestiva de que a mistura fenólica presente no extrato é capaz de sequestrar o radical DPPH em velocidade semelhante a antioxidantes clássicos como o α-tocoferol. Sendo assim, este estudo mostra que o EHA das cascas da Himatanthus drasticus possui importante ação analgésica, anti-inflamatória, cicatrizante e antioxidante. Ademais, este estudo contribui para a validação do uso popular da espécie e abre perspectivas para novas investigações.

The aim of this thesis was to investigate morphogenesis in the fruit fly Drosophila melanogaster using three in vitro tissue culture systems. Primary embryonic cultures derived from Drosophila melanogaster were used to study the effect of the moulting hormone ecdysone on cells in culture. The hypothesis was that the effect of ecdysone on these primary embryonic cells would parallel events which occur during metamorphosis in vivo and therefore the primary embryonic cultures could be used as an ‘in vitro’ model system. Transgenic fly lines expressing GFP were used to visualise and identify specific cell types and it was shown that cells in primary embryonic cultures respond to ecdysone morphologically. However due to the variability of cultures it was concluded that this culture system was not suitable for use as a model system. As defined cell types were observed the development of a protocol suitable for use with the primary embryonic culture system using dsRNA in order to demonstrate RNA interference was undertaken. Although this was unsuccessful, as cells in the primary embryonic cultures appeared to be resistant to dsRNA, some technical avenues remain to be explored. The Drosophila melanogaster cell line, Clone 8+, was used to investigate cell adhesion in tissue culture. Statistical analyses were carried out and it was established that derivatives of the parent cell line, Clone 8+, showed differential adhesion and proliferation characteristics. Analysis of microarray data was carried out in order to identify genes which may be responsible for the loss of cell adhesion in Clone 8+ cell lines and the potential roles of these genes in adhesion were discussed. A gene of interest, glutactin, was identified which may be responsible for loss of cell adhesion. Antibody staining was used to establish the expression of the protein glutactin in the Clone 8+ cell lines. The expression of glutactin suggested that the Clone 8+ cell line had maintained properties of the wing disc epithelial cell-type and disruption of cell polarity was considered as a possible mechanism. It was shown that f-actin colocalised with glutactin and the role of the cytoskeleton in glutactin secretion was discussed. It was concluded that glutactin was not responsible for loss of cell adhesion in the Clone 8+ cell lines. Further analysis of the microarray data revealed potential genes that could be responsible for the loss of cell polarity in the Clone 8+ cell lines and the possibility of cellular senescence was considered. It was hypothesised that the properties of adhesion and proliferation related to their ‘in vitro’ age. In the final investigation the movement of epithelial cells in Drosophila melanogaster third instar larval imaginal discs during morphogenesis was investigated. Firstly a lumen was identified in fixed imaginal disc tissue in association with cells expressing f-actin. This result was discussed in relation to the process of dorsal closure and wound healing. Further investigations involved live imaging of the dynamic process of evagination in the imaginal wing disc using transgenic flies expressing moesin-GFP. It was concluded that the lumen was not associated with the process of wound healing and it was concluded that the lumen appeared to be the mechanism directing peripodial epithelium contraction during morphogenesis of the imaginal wing disc. Dorsal closure and the process of invagination in relation to morphogenesis of the imaginal wing disc were discussed.