Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-12-12T13:41:02Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Alisson_Disssertação_Final_2015.pdf: 4745802 bytes, checksum: 6e96787b9b69ca08b2dfe6e99ae18b3f (MD5) / Made available in DSpace on 2016-12-12T13:41:02Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Alisson_Disssertação_Final_2015.pdf: 4745802 bytes, checksum: 6e96787b9b69ca08b2dfe6e99ae18b3f (MD5) Previous issue date: 2015-08-13 / CAPES / FACEPE / CNPQ / Morus alba L. (amoreira branca) é uma planta utilizada em todo o mundo no tratamento de diversas enfermidades, incluindo distúrbios inflamatórios. Essa planta tem origem asiática, mas demonstrou boa adaptação ao solo brasileiro. Diante da expansão do uso medicinal de M. alba, inclusive comercialmente, o presente trabalho teve por objetivos construir um painel de toxicidade para extrato etanólico das folhas dessa planta e determinar seu potencial como agente antimicrobiano e anti-inflamatório. O extrato foi analisado quanto à composição química por cromatografia em camada delgada. As análises toxicológicas realizadas foram: ensaio de letalidade para o microcrustáceo Artemia salina (concentrações: 100–1000 μg/mL); ensaio de toxicidade aguda por via oral para camundongos (doses: 300 e 2000 mg/kg); determinação da genotoxicidade in vivo para camundongos através do teste do micronúcleo (doses: 75, 150 e 300 mg/kg v.o.); e ensaios de citotoxicidade para as células tumorais humanas NCI-H292, HEp-2, HT-29, MCF-7 e HL-60 (concentração: 50 μg/mL). Na avaliação do potencial anti-inflamatório, foi utilizado o modelo de inflamação aguda induzida por carragenina em bolsão de ar (doses: 75, 150 e 300 mg/kg v.o.). A ação anti-inflamatória foi avaliada através da quantificação do número de leucócitos nos exsudatos coletados dos bolsões. Atividade antimicrobiana foi avaliada contra bactérias e fungos de importância médica. A análise fitoquímica revelou a presença de cumarinas, flavonoides, taninos e triterpenos no extrato, o qual não promoveu mortalidade dos náuplios de A. salina. No teste de toxicidade aguda, não houve mortalidade nem alterações comportamentais nos camundongos tratados com o extrato durante 14 dias. Contudo, a análise de parâmetros hematológicos e bioquímicos revelou que os animais que receberam a dose mais alta apresentaram valores significativamente (p < 0,05) menores de volume corpuscular médio eritrocitário (VCM) e concentração média de hemoglobina corpuscular (CMHC), bem como atividade aumentada de fosfatase alcalina no plasma. Nos tratamentos com o extrato a 300 e 2000 mg/kg, houve redução no número de leucócitos, com diminuição da percentagem de linfócitos e aumento na proporção de células segmentadas. Análise histopatológica dos órgãos dos animais tratados com o extrato a 2000 mg/kg revelou turgidez dos túbulos contorcidos renais, presença de infiltrado leucocitário ao redor da veia centrilobular hepática e elevada dispersão da polpa branca esplênica. Avaliação do potencial genotóxico in vivo revelou que o extrato não causou aumento na frequência de micronúcleos, em comparação com o controle negativo. O extrato também não demonstrou citotoxicidade para as linhagens celulares testadas. Com base nos resultados obtidos nos testes de toxicidade, o potencial anti-inflamatório do extrato foi avaliado utilizando doses menores e igual a 300 mg/kg. Foi verificada redução do número de leucócitos nos grupos tratados com 75 (56,8%), 150 (62,9%) e 300 (65,6%) mg/kg, em comparação com o grupo controle. A dose de 300 mg/kg mostrou efeito de inibição similar ao observado no grupo padrão, que recebeu indometacina a 10 mg/kg, i.p (64,7%). Por fim, o extrato apresentou atividade antimicrobiana contra Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Candida krusei, Candida tropicalis e Aspergillus flavus. Em conclusão, o extrato etanólico de folhas de M. alba apresentou potencial anti-inflamatório e antimicrobiano, não sendo letal nem apresentando efeito genotóxico para camundongos quando ingerido na dose de 300 mg/kg. Contudo, a utilização do extrato na dose de 2000 mg/kg deve ser evitada, devido às alterações bioquímicas, hematológicas e histopatológicas detectadas no ensaio de toxicidade aguda. / Morus alba L. (white mulberry) is a plant used around the world for treatment of several infirmities, including inflammatory disorders. This plant is originated from Asia but showed good adaptation to Brazilian soil. In view of the expansion of the medicinal use of M. alba, including commercially, the present work aimed to build a toxicity panel for an ethanolic extract from leaves of this plant and to determine its potential as antimicrobial and anti-inflammatory agents. The extract was analyzed for chemical composition through thin layer chromatography. The toxicological analyses performed were: lethality assay with the microcrustacean Artemia salina (concentrations: 100–1000 μg/mL); acute oral toxicity assay to mice (doses: 300 and 2000 mg/kg); determination of in vivo genotoxicity to mice by micronucleus test (doses: 75, 150 and 300 mg/kg per os); and cytotoxicity assays to the human tumor cells NCI-H292, HEp-2, HT-29, MCF-7 and HL-60 (concentration: 50 μg/mL). In the evaluation of anti-inflammatory potential, it was used the model of acute inflammation induced by carrageenan in air pouch (doses: 75, 150 and 300 mg/kg per os). The anti-inflammatory action was evaluated by the quantification of the number of leukocytes in exudates collected from the pouches. Antimicrobial activity was evaluated against bacteria and fungi of medical importance. The phytochemical analysis revealed the presence of coumarins, flavonoids, tannins and triterpenes in the extract, which did not promote mortality of A. salina nauplii. In the acute oral toxicity assay, there was no mortality or behavioral alterations in mice treated with the extract during 14 days. However, the analysis of hematologic and biochemical parameters revealed that animals that received the highest dose showed significantly (p < 0.05) lower values of mean erythrocyte corpuscular volume (MCV) and mean concentration of corpuscular hemoglobin (MCHC) as well as increased activity of alkaline phosphatase in plasma. In the treatments with the extract at 300 and 2000 mg/kg, there was reduction in the number of leukocytes, with decrease of lymphocyte percentage and increase in the proportion of segmented cells. Histopathological analysis of the organs from animals treated with the extract at 2000 mg/kg revealed turgidity of renal contorted tubules, presence of leukocyte infiltration around the hepatic centrilobular vein and high dispersion of the spleen white pulp. Evaluation of the in vivo genotoxic potential revealed that the extract did not cause increase in the frequency of micronuclei, in comparison with negative control. The extract also did not show cytotoxicity to the cell lines tested. Based on the results obtained in the toxicity assays, the anti-inflammatory potential of the extract was evaluated using doses lower and equal to 300 mg/kg. It was verified a reduction in the number of leukocytes in the groups treated with the extract at 75 (56.8%), 150 (62.9%) and 300 (65.6%) mg/kg, in comparison with control group. The dose of 300 mg/kg showed inhibitory effect similar to that observed in the standard group, which received indometacin 10 mg/kg, i.p (64.7%). Finally, the extract showed antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Candida krusei, Candida tropicalis and Aspergillus flavus. In conclusion, the ethanolic extract from M. alba leaves showed anti-inflammatory and antimicrobial potential, being not lethal and showing no genotoxic effect when ingested by mice at the dose of 300 mg/kg. However, the use of the extract at 2000 mg/kg should be avoided, due to the biochemical, hematological and histopathological alterations detected in the acute toxicity assay.

L'objectif est de mieux connaître les diverses propriétés (physico-mécaniques, biologiques et chimiques) du bois de Mûrier blanc (Morus alba L.), en tant que matériau éminemment utilisé dans la fabrication de luths Iraniens. La différence de caractéristiques vibratoires de cette espèce a été mesurée en fonction de la présence d'extractibles, mais aussi pour trois traitements artisanaux qui ont été adaptés à l'échelle du laboratoire.Le bois Mûrier blanc est caractérisé par un module spécifique modéré, un amortissement plus bas que prévu et une anisotropie faible entre ses trois axes. Il est précisé que cette espèce ne peut pas être classifiée avec les mêmes normes que les autres bois utilisés pour les instruments classiques Européens. En utilisant les deux extractions indépendantes et successives, il est révélé que l'amortissement est régi par deux types d'extraits, certains l'abaissent et certains l'augmentent. Ces composés ne sont pas tous extractibles par les mêmes solvants. L'immersion de longue durée dans une eau à température ambiante ne modifie pas les propriétés mécaniques de l'espèce, toutefois, elle peut laisser le bois avec des défauts irréversibles. Un court traitement à l'eau chaude (70°C) entraîne l'augmentation de l'amortissement et la diminution du module spécifique. Dans ce dernier cas, des modifications sont presque progressives avec le temps.Le bois de Mûrier blanc semble être « très durable » par rapport à une attaque fongique, même dans des conditions extrêmes. Le délavage dans l'eau n'a pas d'effets sur la résistance de cette espèce contre basidiomycètes. Ce bois est classé « moyennement durable » contre l'attaque des termites et devient sensible après le délavage. Ses extractibles ont donc des effets toxique sur les termites. Les phénols, acides gras, stérols, hydrocarbures supérieurs, et composés aromatiques sont les composés présents dans les extraits de cette espèce. Le résorcinol, composé prédominant trouvé dans l'espèce, a été récemment rapporté comme la cause de plusieurs problèmes de santé, ce qui est aujourd'hui observé chez les artisans Iraniens. / This work aims at studying the several divers properties (Physico-mechanical, biological and chemical) of white Mulberry (Morus alba L.), as the leading material used in fabrication of Iranian lutes. The vibrational characteristic of this species is measured in relation with secondary metabolites (extractives) as well as three artisanal hygro-thermal treatments adapted to laboratory scale.White Mulberry wood is characterized with a moderate specific modulus, less than expected damping and a low anisotropy between three directions. It is specified that this species cannot be described with the same standards as other woods used in European classical instruments. Using independent and successive extractions reveal that two types of extraneous compounds govern the damping in this wood, some raising it, when the rest have a decreasing effect. These compounds are not all extractable by the same solvents.Long time immersion in water at ambient temperature hardly changes mechanical properties of the species, however, it can leave wood with irreversible defects. Hot water treatment at 70°C results for damping and moduli to be increased and decreased respectively. These changes are time related. Several mild desorption and adsorption cycles, seem to reduce damping without greatly endangering the moduli. White Mulberry wood is found to be very durable towards fungi, even in extreme conditions. Water leaching seems to be affectless on this species resistance towards basidiomycetes. This wood is rated moderately durable towards termites and becomes sensible after water leaching. Extractives seem to play important role in this wood natural resistance, as they are found to have toxic effect on termites. Resorcinol, a phenol, is the leading compound in the extractives of white Mulberry. Fatty acids, sterols, higher hydrocarbons, and aromatic compounds are also found as the constituents of secondary metabolites. Resorcinol is documented to cause irritations close to what is normally experienced by the fabricants working with white Mulberry for a long time.