?rea de concentra??o: Farmacologia de produtos naturais e plantas medicinais. / Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-01-07T13:08:56Z No. of bitstreams: 2 michaelle_geralda_santos.pdf: 2124437 bytes, checksum: 4efd2da996375e9bd402d5e8df0ae10f (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-01-07T13:10:40Z (GMT) No. of bitstreams: 2 michaelle_geralda_santos.pdf: 2124437 bytes, checksum: 4efd2da996375e9bd402d5e8df0ae10f (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-01-07T13:11:27Z (GMT) No. of bitstreams: 2 michaelle_geralda_santos.pdf: 2124437 bytes, checksum: 4efd2da996375e9bd402d5e8df0ae10f (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Made available in DSpace on 2015-01-07T13:11:27Z (GMT). No. of bitstreams: 2 michaelle_geralda_santos.pdf: 2124437 bytes, checksum: 4efd2da996375e9bd402d5e8df0ae10f (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Previous issue date: 2014 / A Eriosema crinitum (Kunth) G. Don ? utilizada no Vale do Jequitinhonha como planta anti-inflamat?ria na forma de decoc??o de suas ra?zes. Tendo em vista a complexidade do processo inflamat?rio, o qual envolve a ativa??o de c?lulas do sistema imune e produ??o de diversos mediadores, o objetivo desse estudo foi realizar uma triagem fitoqu?mica e avaliar a poss?vel atividade anti-inflamat?ria, in vitro, do extrato diclorometano-etan?lico de ra?zes de E. crinitum, atrav?s da avalia??o do efeito do mesmo sobre a prolifera??o de linf?citos, a produ??o de citocinas e da ciclooxigenase 2 (COX-2). A triagem fitoqu?mica do extrato foi realizada por meio de rea??es cromog?nicas, fluorog?nicas e de precipita??o, seguidas da an?lise em cromatografia l?quida de alta efici?ncia acoplada a detector de arranjo de diodos (CLAE-DAD). A citotoxicidade do extrato sobre hem?cias, ap?s 4 horas de cultura, e sobre leuc?citos do sangue perif?rico humano, ap?s 24 horas ou 5 dias de cultura, foi avaliada por meio do teste de atividade hemol?tica e pelo m?todo de exclus?o com azul de Tripan, respectivamente. Na avalia??o do efeito do extrato sobre a prolifera??o de linf?citos, por citometria de fluxo, c?lulas mononucleares do sangue perif?rico humano (PBMC) foram incubadas, por 5 dias, em meio de cultura contendo o mit?geno Fitohemaglutinina (PHA), na presen?a ou aus?ncia do extrato nas concentra??es de 25 ?g/mL, 12,5 ?g/mL e 6,25 ?g/mL. Essas mesmas culturas foram realizadas no tempo de 36 horas para an?lise da citocina IL-2, por ELISA. Para an?lise da produ??o das citocinas TNF-? e IFN-?, culturas de sangue total foram estimuladas com Miristato Acetato de Forbol (PMA) e tratadas com o extrato nas concentra??es de 50 ?g/mL, 25 ?g/mL e 12,5 ?g/mL. Essas concentra??es foram utilizadas em culturas de sangue total estimuladas com lipopolissacar?deo (LPS) para investiga??o do efeito do extrato sobre a express?o de COX-2. A produ??o de citocinas e de COX-2 foi analisada por citometria de fluxo. Os resultados evidenciaram a presen?a de terpenos e das subclasses dos flavonoides: flavonas e flavon?is no extrato. O extrato, nas concentra??es iguais ou inferiores a 50 ?g/mL, n?o foi t?xico para culturas celulares de 24 horas e, para culturas de 5 dias, as concentra??es n?o t?xicas foram iguais ou inferiores a 25 ?g/mL. Quanto ? a??o anti-inflamat?ria, o extrato n?o inibiu a produ??o das citocinas TNF-? e IFN-? e tamb?m n?o foi capaz de reduzir a express?o de COX-2. No entanto, o extrato inibiu a prolifera??o de linf?citos e suas subpopula??es T CD4 e TCD8, tendo uma efic?cia em rela??o ? Dexametasona de 71,65%, 53,14% e 167,94% para linf?citos totais, T CD4 e T CD8, respectivamente. O extrato tamb?m reduziu os n?veis de IL-2 nas culturas estimuladas com PHA. Esses achados sugerem que o extrato apresenta um efeito inibidor, principalmente sobre a prolifera??o de c?lulas T CD8, associado ? inibi??o na produ??o de IL-2. Diante dos resultados apresentados, os princ?pios ativos presentes na planta parecem exercer suas fun??es anti-inflamat?rias regulando a prolifera??o de linf?citos T, principalmente os linf?citos T CD8. Essa fun??o poderia ser explorada em desordens de natureza inflamat?ria-linfoproliferativa, bem como preven??o da rejei??o de transplantes de ?rg?os. Para tal, ensaios adicionais s?o necess?rios para investigar e identificar os constituintes ativos e os mecanismos moleculares envolvidos na a??o inibit?ria apresentada pelo extrato. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2014. / ABSTRACT The Eriosema crinitum (Kunth) G. Don is used in Jequitinhonha Valley as anti-inflammatory plant in the form of decoction of its roots. Given the complexity of the inflammatory process, which involves the activation of immune cells and production of several mediators, the aim of this study was to perform a phytochemical screening and evaluate the potential anti-inflammatory activity, in vitro, the dichloromethane-ethanol extract of the roots of E. crinitum by assessing the effect of the same on lymphocyte proliferation, production of cytokines and cyclooxygenase-2 (COX-2). The phytochemical screening of the extract was performed using chromogenic reactions, fluorogenic and precipitation followed by analysis in high efficiency liquid chromatography coupled with a diode array detector (HPLC-DAD). The cytotoxicity of the extract after 24 hours or 5 days of culture on erythrocytes and leukocytes in human peripheral blood was evaluated by the test of hemolytic activity and the method of exclusion with Trypan blue, respectively. In evaluating of the effect of the extract on the proliferation of lymphocytes by flow cytometry, mononuclear cells from human peripheral blood (PBMC) have been incubated for 5 days in a culture medium containing the mitogen Phytohemagglutinin (PHA) in the presence or absence of extract at concentrations of 25 ?g/mL, 12.5 ?g/mL and 6.25 ?g/mL. These cultures too were performed in the time of 36 hours for analysis of IL -2 by ELISA. To analyze the production of cytokines TNF-? and IFN-?, whole blood cultures were stimulated with Phorbol Myristate acetate (PMA) and treated with the extract at concentrations of 50 ?g/mL, 25 ?g/mL and 12.5 ?g/mL. These concentrations were used in whole blood cultures stimulated with lipopolysaccharide (LPS) to investigate the effect of the extract on the expression of COX-2. Cytokine production and COX-2 was analyzed by flow cytometry. The results showed the presence of terpenes and subclasses of flavonoids: flavones and flavonols in extract. The extract, in concentration equal or greater than 50 ?g/mL was not toxic to cell cultures after 24 hours and in culture after 5 days, non- toxic concentrations were equal to or greater than 25 ?g/mL. In relation to the anti-inflammatory action, the extract did not inhibit the production of TNF-? and IFN-? and also was not able to reduce the expression of COX-2. Moreover, the extract inhibited the proliferation of lymphocytes and their subpopulations CD4 and CD8, and its efficacy compared to dexamethasone was 71.65%, 167.94% and 53.14% for total lymphocytes, CD4 and CD8, respectively. The extract also reduced the levels of IL-2 in cultures stimulated with PHA. These findings suggest that the extract has an inhibitory effect, especially on the proliferation of CD8 T cells associated with the inhibition of IL-2. Considering the presented results, the active molecules present in the plant seem to play their anti-inflammatory functions by regulating the T lymphocytes proliferation,?mainly T CD8 lymphocytes. This function could be exploited in inflammatory lymphoproliferative disorders, as well as in the prevention of transplant rejection. Further trials are needed to investigate the active constituents and the molecular mechanisms involved in the inhibitory action displayed by the extract.

Submitted by PPG Medicina e Ci?ncias da Sa?de (medicina-pg@pucrs.br) on 2018-06-22T19:59:08Z No. of bitstreams: 1 CLARICE_MARTINS_FEYH_BIRNFELD.pdf: 3318155 bytes, checksum: aa5476b4fe73fb5e7e0b2678aef02e4b (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-06-29T12:31:16Z (GMT) No. of bitstreams: 1 CLARICE_MARTINS_FEYH_BIRNFELD.pdf: 3318155 bytes, checksum: aa5476b4fe73fb5e7e0b2678aef02e4b (MD5) / Made available in DSpace on 2018-06-29T12:38:48Z (GMT). No. of bitstreams: 1 CLARICE_MARTINS_FEYH_BIRNFELD.pdf: 3318155 bytes, checksum: aa5476b4fe73fb5e7e0b2678aef02e4b (MD5) Previous issue date: 2014-10-31 / Objectives: To evaluate the expression of Hsp27 by immunohistochemistry in tissue samples of invasive ductal carcinoma in women with and without the presence of metastasis in sentinel axillary lymph node compared to samples from patients diagnosed with fibrocystic changes in breast tissue. In addition, to study the relationship between the rate of cell proliferation and apoptosis by evaluating the expression of Ki-67 protein and caspase-3 by immunohistochemistry. Material and Methods: A cross- sectional study was designed to select patients treated at Hospital S?o Lucas of PUCRS between September 2001 and October 2009. The patients were divided in three groups: 25 patients diagnosed with invasive ductal carcinoma of breast with axillary lymph node metastasis, 25 patients with invasive ductal carcinoma of breast without axillary lymph node metastasis and a control group composed by tissue samples of 25 patients diagnosed with fibrocystic change of the breast. The expression of Hsp27, Ki-67 and caspase 3 expression was evaluated by image analysis and the rate of proliferation (Ki-67) and apoptosis (caspase 3) was calculated for the three groups. Results: The present study showed an increased expression of Hsp27 in the group of invasive ductal carcinomas without metastasis as compared to the control group. There was no significant difference of expression of Hsp27 between the group of ductal carcinoma with axillary metastasis and the control group. The expression of Hsp27 was significantly higher in the group without axillary metastasis as compared to the group with metastasis in both the primary tumor (<0.001) and the respective lymph node tissue samples (=0.003). The expression of Ki-67 and the rate of proliferation (ki-67) and apoptosis (caspase 3) were significantly higher in the groups with invasive ductal carcinoma as compared to the control group. There was no difference in the expression of Ki-67 and the rate of proliferation and apoptosis between the two groups of invasive ductal carcinoma. There was no significant difference in the expression of caspase 3 among the three groups. Conclusion: The quantitative analysis of Hsp27 demonstrated increased expression of this protein in carcinomas without axillary metastases, with a significantly higher expression in the primary tumor and lymph node tissue samples of this group as compared to the group without axillary metastasis. Although the methodology employed and the number of cases evaluated in this work do not allow one to conclude that such behavior in metastatic ductal carcinomas of breast is constant, such finding warrants further investigation. / Objetivos: Avaliar a express?o de Hsp27 (HspB1) em amostras teciduais de carcinomas ductais invasores em mulheres com e sem presen?a de met?stase em linfonodo sentinela axilar e estudar a rela??o da taxa de prolifera??o celular e apoptose, atrav?s da avalia??o imunoistoqu?mica da express?o das prote?nas Hsp27, Ki-67 e caspase 3 em carcinomas ductais invasores de mulheres com e sem presen?a de met?stase linfonodal, tratadas em um hospital de ensino, comparado com amostras de pacientes diagnosticadas com altera??o fibroc?stica no tecido mam?rio. Material e M?todos: Foi realizado um estudo do tipo transversal, sendo selecionadas para este trabalho pacientes tratadas no Hospital S?o Lucas da PUCRS no per?odo entre setembro de 2001 e outubro de 2009 cujas bi?psias foram divididas em tr?s grupos: 25 pacientes com diagn?stico de carcinoma ductal invasor de mama com presen?a de met?stase linfonodal; e 25 pacientes com carcinoma ductal invasor de mama sem presen?a de met?stase linfonodal. Para constitui??o do grupo controle foi realizada a sele??o aleat?ria de 25 pacientes com diagn?stico de altera??o fibroc?stica da mama. Atrav?s da t?cnica de imunoistoqu?mica, tamb?m foram avaliadas as express?es de Hsp27, caspase 3 e Ki67 por an?lise de imagem digital e calculada a raz?o entre a taxa de prolifera??o (Ki-67) e apoptose (caspase 3) dos grupos estudados. Resultados: O presente estudo evidenciou a express?o aumentada da Hsp27 em carcinomas ductais invasores sem met?stase axilar, quando comparados ao grupo dos carcinomas ductais invasores com met?tase axilar e grupo controle. A express?o de Hsp 27 foi maior no grupo com carcinoma invasor sem met?stase axilar tanto no tecido da neoplasia prim?ria (p<0,001) quanto no respectivo tecido linfonodal comparado ao grupo com carcinoma com met?stase axilar (=0,003). Houve diferen?a na express?o da prote?na Ki-67 e raz?o entre prolifera??o celular e apoptose entre os grupos de pacientes com carcinoma ductal invasor com e sem met?stases quando comparados ao grupo controle, por?m n?o houve diferen?a significativa da express?o de Ki-67 e da raz?o entre prolifera??o e apoptose entre os grupos com carcinoma invasor com e sem met?stase, ou entre a express?o de caspase 3 nos tr?s grupos quando comparados entre si. Conclus?o: A an?lise quantitativa da Hsp27 demonstrou maior express?o da prote?na nos tecidos da neoplasia prim?ria e nos linfonodos em carcinomas sem met?stase em linfonodo sentinela axilar. Embora a metodologia empregada e o n?mero de casos estudados nesse trabalho n?o permitam concluir que esse comportamento na doen?a metast?tica seja um achado constante, esse resultado justifica a necessidade de avaliar esse achado em estudos futuros.

?rea de concentra??o: Ci?ncias Fisiol?gicas. / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-01-09T16:47:50Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lucas_abreu_costa.pdf: 2675995 bytes, checksum: b6686da3b0509ff9d2b21c38e1fdc5ce (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-01-10T13:46:07Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lucas_abreu_costa.pdf: 2675995 bytes, checksum: b6686da3b0509ff9d2b21c38e1fdc5ce (MD5) / Made available in DSpace on 2017-01-10T13:46:07Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lucas_abreu_costa.pdf: 2675995 bytes, checksum: b6686da3b0509ff9d2b21c38e1fdc5ce (MD5) Previous issue date: 2016 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A planta Ageratum fastigiatum (Gardn.) R. M. King et H. Rob. ? comumente utilizada na medicina popular da regi?o do Vale do Jequitinhonha como anti-inflamat?rio e analg?sico. Em virtude do papel do sistema imune e de fatores inflamat?rios sol?veis na etiologia e no mecanismo fisiopatol?gico de diversas doen?as inflamat?rias, este trabalho teve como objetivo investigar par?metros relacionados ? atividade anti-inflamat?ria do extrato etan?lico de A. fastigiatum e de uma fra??o derivada desse extrato. A identifica??o dos constituintes qu?micos da fra??o do extrato etan?lico de A. fastigiatum foi realizada por meio da Cromatografia L?quida de Alta Efici?ncia com Detectores de Arranjo de Diodo acoplada ? espectrometria de massas (CLAE-DAD-MS) seguida pela Espectrometria de Massas em Tandem com ioniza??o por electrospray (ESI-MS/MS). A t?cnica de exclus?o com azul de tripan foi utilizada para determina??o da viabilidade das culturas de c?lulas mononucleares do sangue perif?rico (PBMC) incubadas por 8, 24 e 120 horas com o extrato etan?lico, a fra??o e o solvente Dimetilsulf?xido (DMSO). Na avalia??o do perfil proliferativo de linf?citos pela t?cnica de citometria de fluxo, PBMC estimuladas com o mit?geno Fitohemaglutinina (PHA) foram incubados por 120 horas na aus?ncia ou presen?a de diferentes concentra??es n?o t?xicas dos componentes citados. Para an?lise da produ??o das citocinas pr?-inflamat?rias, PBMC tratadas ou n?o com diferentes concentra??es n?o t?xicas do extrato etan?lico de A. fastigiatum, da fra??o e do solvente DMSO foram incubadas por 8 horas e estimulada nas ?ltimas 4 horas com Miristato Acetato de Forbol (PMA), utilizando a t?cnica de citometria de fluxo, linf?citos totais, c?lulas CD4+ e CD8+ foram avaliados quanto ? produ??o de IFN-?, TNF-? e IL-2. De acordo com nossos resultados, a concentra??o de DMSO, para todas as culturas celulares tratadas com o extrato etan?lico ou sua fra??o, foi definida como sendo 1% v/v para avalia??o da citotoxicidade e an?lise da produ??o de citocinas em linf?citos e de 0.5% v/v no ensaio de prolifera??o celular. Na an?lise fitoqu?mica da fra??o do extrato etan?lico foram identificados dois flavon?is nunca antes descrito na constitui??o qu?mica da planta, a 7-metil-quercetina e a Rhamnocitrina, tais compostos n?o alteraram os par?metros anti-inflamat?rios avaliados neste estudo. O extrato etan?lico, por sua vez, nas concentra??es de 6,25 e 12,5 ?g/mL reduziu significativamente a prolifera??o de linf?citos, ao mesmo tempo, nas concentra??es de 50 e 75 ?g/mL inibiu a produ??o das citocinas TNF-? e IL-2 em c?lulas CD8+, na maior concentra??o inibiu ainda a produ??o de IL-2 na popula??o de linf?citos totais. Sugere-se que o extrato etan?lico de A. fastigiatum possui componentes ativos agindo sinergicamente ou de forma isolada que contribuem para atividade anti-inflamat?ria atribu?da ? planta em seu uso na medicina popular. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Ageratum fastigiatum (Gardner.) R. M. King et H. Rob. is a plant commonly used in the folk medicine of the Jequitinhonha Valley as anti-inflammatory and analgesic. Because of the role of the immune system and soluble inflammatory factors in the etiology and pathophysiological mechanism of various inflammatory diseases, this study aimed to investigate parameters related to the anti-inflammatory activity of the ethanol extract of A. fastigiatum and a fraction derived from this extract. The identification of the chemical constituents of the fraction of ethanol extract of A. fastigiatum was performed by High Performance Liquid Chromatography with Efficiency Diode Arrangement detectors coupled to mass spectrometry (HPLC-DAD-MS) followed by Mass Spectrometry in Tandem with electrospray ionization (ESI-MS/MS). The exclusion technique with Trypan blue was used to determine the viability of peripheral blood mononuclear cell (PBMC) cultures, previously incubated for 8, 24 and 120 hours with the ethanol extract, the fraction and the solvent dimethylsulfoxide (DMSO). In the evaluation of the proliferative profile of lymphocytes using the flow cytometry technique, PBMC stimulated with the mitogen Phytohemagglutinin (PHA) were incubated for 120 hours in the absence or presence of different non-toxic concentrations of the mentioned components. In order to analyze the production of pro-inflammatory cytokines, PBMC treated or not with different non-toxic concentrations of the ethanolic extract of A. fastigiatum, of the fraction, and of the solvent DMSO was incubated for 8 hours and stimulated with phorbol myristate acetate (PMA) in the last 4 hours. Subsequently, total lymphocytes, CD4+ and CD8+ cells were assessed for production of IFN-?, TNF-? and IL-2, using flow cytometry. According to our results, the concentration of DMSO, for all cell cultures treated with ethanolic extract and its fraction was defined as 1% v/v for evaluation of cytotoxicity and analysis of cytokine production by lymphocytes, and 0.5% v/v to be used in the cell proliferation assay. In the phytochemical analysis of the fraction of ethanol extract, were identified two flavonols never before described in the chemical constitution of the plant, 7-methyl-quercetin and Rhamnocitrina. Such compounds did not modified the anti-inflammatory parameters evaluated in this study. The ethanol extract, on the other hand, in concentrations of 6.25 and 12.5 ?g/ml significantly reduced the proliferation of lymphocytes, while at concentrations of 50 and 75 ug/mL, it inhibited the production of TNF-? and IL-2 in CD8+, in addition, at the highest concentration it inhibited IL-2 production by total lymphocytes population. It is suggested that the ethanol extract of A.fastigiatum has active components acting synergistically or separately, resulting in the anti-inflammatory activity attributed to the plant by the folk medicine.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-01-05T16:59:19Z No. of bitstreams: 1 GeissyLainnyDeLimaAraujo_DISSERT.pdf: 3295602 bytes, checksum: 5e99ad6700c5552b3b1ad3651c7bce8c (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-01-08T18:10:33Z (GMT) No. of bitstreams: 1 GeissyLainnyDeLimaAraujo_DISSERT.pdf: 3295602 bytes, checksum: 5e99ad6700c5552b3b1ad3651c7bce8c (MD5) / Made available in DSpace on 2016-01-08T18:10:33Z (GMT). No. of bitstreams: 1 GeissyLainnyDeLimaAraujo_DISSERT.pdf: 3295602 bytes, checksum: 5e99ad6700c5552b3b1ad3651c7bce8c (MD5) Previous issue date: 2014-10-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / O Sonic Hedgehog (Shh) ? um morf?geno com importantes a??es no sistema nervoso central (SNC) em desenvolvimento, assim como na vida adulta em quadros de les?o tecidual e processos tumorig?nicos. A rela??o da sua via de sinaliza??o com prolifera??o, diferencia??o e sobreviv?ncia celular ? amplamente estudada em regi?es ventrais do SNC. No entanto, o papel da sinaliza??o por Shh em egi?es dorsais, como o telenc?falo dorsal, origem do c?rtex cerebral, n?o est? bem documentada. A partir do cultivo de c?lulas de roedores retiradas do telenc?falo dorsal em desenvolvimento, observamos a influ?ncia do Shh sobre a prolifera??o e diferencia??o das c?lulas-tronco neurais. Utilizando v?deo-microscopia de tempo intervalado, podemos avaliar o tempo de ciclo celular, tamanho de c?lulas progenitoras antes da divis?o celular e tipo de divis?o sofrida pelas c?lulas na presen?a ou na aus?ncia de sinaliza??o por Shh. Verificamos um aumento do n?mero de c?lulas em estado proliferativo assim como um aumento de c?lulas reativas para o marcador astrocit?rio GFAP com o tratamento com Shh. Em contrapartida, ap?s bloqueio da sinaliza??o por Shh, observamos um menor n?mero de c?lulas em estado proliferativo, desacelera??o do ciclo celular, aumento da morte celular e redu??o da astrogliog?nese. Por fim, com intuito de avaliar a influencia do Shh in vivo, n?s injetamos f?rmacos agonista (Purmorfamina) e antagonista (Ciclopamina) da via de sinaliza??o dessa prote?na em diferentes per?odos da gesta??o de roedores. Ao avaliar os animais na vida p?s-natal, observamos um aumento no n?mero de progenitores gliais gerados com o tratamento com Purmorfamina na subst?ncia branca, enquanto na subst?ncia cinzenta n?o parece haver altera??o dessa popula??o em ambos os tratamentos. Al?m disso, a popula??o de c?lulas astrocit?rias, evidenciada por marcadores espec?ficos, parece estar alterada com a manipula??o da sinaliza??o por Shh. Em conjunto, nossos dados sugerem que a Shh est? presente no telenc?falo dorsal em per?odos precoces do desenvolvimento e influencia a gera??o, sobreviv?ncia e prolifera??o de progenitores e c?lulas gliais.

Made available in DSpace on 2014-12-17T14:03:44Z (GMT). No. of bitstreams: 1 HugoWBA_DISSERT.pdf: 1563778 bytes, checksum: ea0b3bf4e13c36f81c11450e423c8df4 (MD5) Previous issue date: 2014-02-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The alginic acid or alginates are acidic polysaccharides found in brown seaweed widely used in food, cosmetic, medical and pharmaceutical industry. This paper proposes the extraction, chemical characterization and verification of the pharmacological activities of brown seaweed variegata Lobophora . The alginate was extracted from the seaweed Lobophora variegata and part was sulphated for comparative purposes. The native extract showed 42% total sugar, 65% uronic acid, 0,36 % protein and 0% of sulfate, while the sulfate showed 39% , 60%, 0.36% and 27,92 % respectively. The presence of a sulfate group may be observed by the metachromasia with toluidine blue in electrophoresis system and characteristic vibration 1262,34 cm-1 in infrared spectroscopy connections assigned to S = O. We observed the formation of films and beads of native alginate, where more concentrated solution 6% resulted in a thicker and more consistent film. Native alginate showed proliferative activity at concentrations (25 and 50 mcg), (50 mg) and (100 mg) in 3T3 cell line in 24h, 48h and 72h, respectively , as the sulfated (100 mg) in 24 . Also showed antiproliferative or cytotoxic activity in HeLa cells of strain, (25 and 100 mg), (25 and 100 mg) and (25, 50 and 100 mg), to native, now for the sulfate concentrations (100 mg) in 24 (25, 50 and 100 mg) in 48 hours, and (50 and 100 mg ) 72h. For their antioxidant activity, the sulfated alginates have better total antioxidant activity reaching 29 % of the native activity while 7.5 % of activity . For the hydroxyl radical AS showed high inhibition ( between 77-83 % ) in concentrations, but the AN surpassed these numbers in the order of 78-92 % inhibition. The reducing power of AN and AS ranged between 39-82 % . In the method of ferric chelation NA reached 100 % chelating while the AS remained at a plateau oscillating 6.5%. However, in this study , we found alginates with promising pharmacological activities, to use in various industries as an antioxidant / anti-tumor compound / Os ?cidos alg?nicos ou alginatos s?o polissacar?deos ?cidos presentes em algas marrons amplamente utilizadas na ind?stria de alimentos, est?tica, m?dica e farmac?utica. Este trabalho prop?e a extra??o, caracteriza??o qu?mica e verifica??o das atividades farmacol?gicas da alga marinha marrom Lobophora variegata. O alginato foi extra?do da alga Lobophora variegata e parte foi sulfatado para fins comparativos. O extrato nativo apresentou 42% de a??car total, 65% de ?cido ur?nico, 0,36% de prote?na e 0% de sulfato, enquanto a sulfatada apresentou 39%, 60%, 0,36% e 27,92% respectivamente. A presen?a do grupo sulfato pode ser verificada atrav?s da metacromasia com o corante azul de toluidina no sistema de eletroforese e vibra??o caracter?stica em 1262,34 cm-1 na espectroscopia de infravermelho, atribu?do a liga??es S=O. Observou-se a forma??o de filmes e esferas de alginato nativo, onde a solu??o mais concentrada 6%, resultou em um filme mais espesso e consistente. O alginato nativo apresentou atividade proliferativa nas concentra??es (25 e 50?g), (50 ?g) e (100 ?g) em linhagem celular 3T3 em 24h, 48h e 72h, respectivamente, j? o sulfatado em (100 ?g) em 24h. Apresentou tamb?m atividade antiproliferativa ou citot?xica em c?lulas da linhagem HeLa, com (25 e 100 ?g), (25 e 100 ?g) e (25, 50 e 100 ?g), para o nativo, j? para a sulfatada nas concentra??es (100 ?g) em 24h, (25, 50 e 100 ?g) em 48h, e (50 e 100 ?g) 72h. Quanto a atividade antioxidante, os alginatos sulfatados apresentam melhor atividade antioxidante total chegando a 29% de atividade enquanto o nativo 7,5% de atividade. Para o radical hidroxila o AS apresentou alta inibi??o (entre 77-83%) nas concentra??es testadas, por?m o AN superou estes n?meros na ordem de 78-92% de inibi??o. O poder redutor de AN e AS variou entre 39-82%. Na metodologia de quela??o f?rrica, o NA chegou a 100% de quela??o, enquanto o AS permaneceu em um plat? oscilando em 6,5%. Contudo, neste trabalho, pudemos constatar alginatos com promissoras atividades farmacol?gicas, com uso nas diversas ind?strias como um composto antioxidante/antitumoral

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-07-17T13:12:44Z No. of bitstreams: 1 FernandaGinaniAntunes_TESE.pdf: 2184683 bytes, checksum: 53f3cc6dc1251b2dcdc05b91678ae4d3 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-07-18T15:34:49Z (GMT) No. of bitstreams: 1 FernandaGinaniAntunes_TESE.pdf: 2184683 bytes, checksum: 53f3cc6dc1251b2dcdc05b91678ae4d3 (MD5) / Made available in DSpace on 2017-07-18T15:34:49Z (GMT). No. of bitstreams: 1 FernandaGinaniAntunes_TESE.pdf: 2184683 bytes, checksum: 53f3cc6dc1251b2dcdc05b91678ae4d3 (MD5) Previous issue date: 2017-02-23 / O laser de baixa intensidade (LBI) tem apresentado resultados positivos na prolifera??o de diversos tipos celulares in vitro. A primeira parte do trabalho avaliou o efeito do LBI na prolifera??o e viabilidade de c?lulas-tronco da polpa de dentes dec?duos humanos esfoliados (SHED). As c?lulas foram irradiadas ou n?o (controle) com um laser diodo InGaAlP (660 nm, 30 mW, modo de a??o cont?nuo) usando duas diferentes doses (0,5 J/cm2 por 16 segundos e 1,0 J/cm2 por 33 segundos) e os tr?s grupos foram estudados nos intervalos de 0, 24, 48 e 72 h. Foi observado que a dose de 1,0 J/cm2 promoveu um aumento na prolifera??o celular nos intervalos de 48 e 72 h quando comparada ao grupo controle e ? dose de 0,5 J/cm2 e a viabilidade celular n?o foi afetada pela irradia??o ao longo do experimento. Em conjunto, os dados mostraram que os par?metros do LBI utilizados (660 nm, 30 mW, 1,0 J/cm2) promoveram prolifera??o das SHEDs e mantiveram a sua viabilidade. A partir destes resultados favor?veis, a segunda parte do trabalho avaliou o efeito do LBI com os mesmos par?metros na prolifera??o de SHEDs cultivadas sobre arcabou?os nanofibrosos de PLA confeccionados pela t?cnica de eletrofia??o. Este procedimento permite a obten??o de arcabou?os tridimensionais que simulam a matriz extracelular a partir de um biomaterial com propriedades f?sicas favor?veis e baixo custo. Tr?s grupos experimentais (G1 ? n?o irradiado; G2 ? irradiado com 0,5 J/cm2; G3 ? irradiado com 1,0 J/cm2) foram analisados nos intervalos de 24, 48 e 72 h. Os resultados indicaram que o PLA n?o ? citot?xico para as SHEDs e que os grupos submetidos ? laserterapia tiveram maior prolifera??o celular quando comparados com o grupo controle n?o irradiado. Com base nos achados, evidenciou-se que as nanofibras de PLA s?o arcabou?os favor?veis para o cultivo de SHEDs e que a laserterapia estimulou a prolifera??o quando aplicada nas c?lulas cultivadas sobre este arcabou?o. Com base nos dados gerais obtidos, conclui-se que a laserterapia nos par?metros avaliados apresenta efeitos bioestimulat?rios nas SHEDs cultivadas tanto na superf?cie padr?o (pl?stico da placa de cultivo) quanto sobre arcabou?os nanoestruturados de PLA. / The low-level laser therapy (LLLT) has been shown positive results in the in vitro proliferation of several cell types. The first part of the study evaluated the effect of LLLT on the proliferation and viability of stem cells from human exfoliated deciduous teeth (SHED).Cells were irradiated or not (control) with an InGaAlP diode laser (660 nm, 30 mW, continuous action mode) using two different doses (0.5 J/cm2 for 16 seconds and 1.0 J/cm2 for 33 seconds) and the three groups were analyzed at the intervals of 0, 24, 48 and 72 h.It was observed that the dose of 1.0 J/cm2 promoted an increase in cell proliferation at the 48 and 72 h intervals when compared to the control group and the dose of 0.5 J/cm2, and that cell viability was not affected by irradiation throughout the experiment. Taken together, the data showed that the LLLT parameters (660 nm, 30 mW, 1.0 J/cm2) promoted proliferation of SHEDs and maintained their viability. Based on these favorable results, the second part of the study evaluated the effect of LLLT with the same parameters on the proliferation of SHEDs cultured on PLA nanofibrous scaffolds obtained by electrospinning. This procedure allows obtaining three-dimensional scaffolds that mimic the extracellular matrix from a biomaterial with favorable physical properties and low cost.Three experimental groups (G1 - non-irradiated; G2 - irradiated with 0.5 J/cm2; G3 - irradiated with 1.0 J/cm2) were analyzed at 24, 48 and 72 h. The results indicated that PLA is not cytotoxic to SHEDs and that the groups submitted to laser therapy had higher cell growth when compared to the nonirradiated control group. Based on these findings, it was shown that PLA nanofibers are favorable scaffolds for the cultivation of SHEDs and that laser therapy stimulated the proliferation when applied to the cells cultured on this scaffold. Based on the general data obtained, it is concluded that the laser therapy in the evaluated parameters has biostimulatory effects on the proliferation of SHEDs on both the standard surface (plastic of the culture plate) and on nanostructured PLA scaffolds.

Made available in DSpace on 2014-12-17T15:43:54Z (GMT). No. of bitstreams: 1 DiegoMS_DISSERT.pdf: 1792102 bytes, checksum: 305ff686e7a1606e27026b528fd1d091 (MD5) Previous issue date: 2013-01-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in &#945;-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm? - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm? promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm?) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm? exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm?. No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration / O laser de baixa intensidade (LBI) tem sido utilizado na Odontologia com a finalidade de promover cicatriza??o e regenera??o dos tecidos. A literatura mostra um efeito positivo do LBI na prolifera??o celular, por?m pouco se sabe sobre a sua efic?cia na prolifera??o de c?lulas-tronco. O objetivo deste estudo foi avaliar o efeito da irradia??o do LBI na taxa proliferativa de c?lulas -tronco do ligamento periodontal humano. Extratos de ligamento periodontal foram isolados de dois terceiros molares h?gidos removidos por indica? ?o cir?rgica e/ou ortod?ntica. Ap?s a digest?o enzim?tica, as c?lulas foram cultivadas em meio de cultura &#945;-MEM suplementado com antibi?ticos e 15% de soro fetal bovino. No terceiro subcultivo, as c?lulas foram irradiadas com um laser diodo InGaAlP, utilizando-se duas diferentes densidades de energia (0,5J/cm 2 - 16 segundos e 1,0J/cm? - 33 segundos), comprimento de onda de 660nm e pot?ncia de 30mW. Uma nova irradia??o, utilizando os mesmos par?metros, foi realizada 48 h ap?s a primeira. Um grupo controle (n?o irradiado) foi mantido nas mesmas condi??es experimentais de cultivo. O m?todo de exclus?o por azul de Tripan e a atividade mitocondrial das c?lulas medida atrav?s do ensaio de MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetraz?lio], nos intervalos de 0, 24, 48 e 72 h p?s-irradia??o, foram utilizados a fim de avaliar a prolifera??o celular. Os dados das contagens celulares foram submetidos a testes estat?sticos n?o param?tricos de Kruskal-Wallis e Mann-Whitney, considerando um intervalo de confian?a de 95%. Com o objetivo de verificar poss?veis altera??es morfol?gicas nucleares induzidas pelo laser, as c?lulas foram submetidas ? marca??o com DAPI (4 -6-Diamidino-2-phenylindole) no intervalo de 72 h. Os resultados do presente estudo mostraram que a densidade de energia de 1,0 J/cm? promoveu maior prolifera??o das c?lulas em compara??o com os outros grupos (controle e laser 0,5 J/cm?) nos intervalos de 48 e 72 h. A atividade mitocondrial, medida pelo ensaio de MTT, apresentou resultados semelhantes ?s contagem celulares com azul de Tripan, com o grupo irradiado com 1,0 J/cm? exibindo uma atividade significativamente maior do MTT nos intervalos de 48 e 72 h, quando comparado com o grupo irradiado com 0,5 J/cm?. Nenhuma altera??o morfol?gica nuclear foi observada, tanto das c?lulas do grupo controle quanto nas c?lulas irradiadas. Conclui-se que o LBI apresenta efeitos estimulantes sobre a prolifera??o de c?lulas-tronco do ligamento periodontal humano. Portanto, a aplica??o da laserterapia neste tipo celular pode ser importante para futuros avan?os na regenera??o periodontal

Made available in DSpace on 2014-12-17T14:03:37Z (GMT). No. of bitstreams: 1 MariliaSNS_TESE.pdf: 2303324 bytes, checksum: d055f1a32ee94b3ffe5f389dbf9dffe0 (MD5) Previous issue date: 2014-04-08 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Compounds derived from fungi has been the subject of many studies in order to broaden the knowledge of their bioactive potential. Polysaccharides from Caripia montagnei have been described to possess anti-inflammatory and antioxidant properties. In this study, glucans extracted from Caripia montagnei mushroom were chemically characterized and their effects evaluated at different doses and intervals of treatment. It was also described their action on colonic injury in the model of colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS), and its action on cells of the human colon carcinoma (HT-29). Compounds extracted of C. montagnei contain high level of carbohydrates (96%), low content of phenolic compounds (1.5%) and low contamination with proteins (2.5%). The (FT-IR) and (NMR) analysis showed that polysaccharides from this species of mushroom are composed of &#945;- and &#946;-glucans. The colonic damage was evaluated by macroscopic, histological, biochemical and immunologic analyses. The results showed a reduction of colonic lesions in all groups treated with the glucans of Caripia montagnei (GCM). GCM significantly reduced the levels of IL-6 (50 and 75 mg/kg, p < 0.05), a major inflammatory cytokine. Biochemical analyses showed that such glucans acted on reducing levels of alkaline phosphatase (75 mg/kg, p < 0.01), nitric oxide (p < 0.001), and myeloperoxidase (p < 0.001). These results were confirmed microscopically by the reduction of cellular infiltration. The increase of catalase activity suggest a protective effect of GCM on colonic tissue, confirming their anti-inflammatory potential. GCM displayed cytostatic activity against HT-29 cells, causing accumulation of cells in G1 phase, blocking the cycle cell progression. Those glucans also showed ability to modulate the adhesion of HT-29 cells to Matrigel? and reduced the oxidative stress. The antiproliferative activity against HT-29 cells displayed by GCM (p <0.001) can be attributed to its cytostatic activity and induction of apoptosis by GCM / Compostos derivados de fungos tem sido alvo de muitos estudos a fim de desenvolver o conhecimento acerca de seu potencial bioativo. Polissacar?deos de Caripia montagnei j? foram descritos por possu?rem propriedades anti-inflamat?ria e antioxidante. Neste estudo, os polissacar?deos extra?dos do fungo Caripia montagnei foram caracterizados quimicamente e seus efeitos sobre as les?es intestinais foram avaliados em diferentes intervalos de tratamento no modelo de colite induzida por ?cido 2,4,6 - trinitrobenzenossulf?nico (TNBS), verificou-se ainda sua a??o sobre c?lulas do carcinoma de c?lon humano, HT-29. Na an?lise realizada no extrato obtido de C. montagnei foi verificado que este ? formado principalmente, por carboidratos (96%) apresentando um baixo teor de compostos fen?licos (1,5%) e baixa contamina??o prot?ica (2,5%). As an?lises por espectroscopia de infra vermelho (FT-IR) e resson?ncia magn?tica nuclear (RMN) mostraram que os polissacar?deos desta esp?cie de fungo s?o &#945; e &#946; -glucanas. O dano col?nico foi avaliado por an?lises macrosc?picas, histol?gicas, bioqu?micas e imunol?gicas. Os resultados mostraram a redu??o das les?es no c?lon em todos os grupos tratados com as glucanas (GCM). GCM reduziram significativamente os n?veis de IL-6 (50 e 75 mg/Kg, p < 0,05), uma importante citocina inflamat?ria. As an?lises bioqu?micas mostraram que essas glucanas atuaram na redu??o dos n?veis de fosfatase alcalina (75 mg/Kg, p < 0,01), ?xido n?trico (p < 0,001) e mieloperoxidase (p < 0,001). Estes resultados foram confirmados pela redu??o da infiltra??o celular observado microscopicamente. O aumento da atividade da catalase, sugere um efeito protetor de GCM no tecido do c?lon, o que confirma o seu potencial anti-inflamat?rio. GCM mostraram atividade citost?tica sobre as c?lulas HT-29, causando ac?mulo de c?lulas na fase G1 e impedindo, assim, a progress?o do ciclo celular. As glucanas deste estudo tamb?m mostraram habilidade em modular a ades?o de c?lulas HT-29 ao Matrigel? e reduzir o estresse oxidativo nessas c?lulas. A atividade antiproliferativa contra c?lulas HT-29 exibida por GCM pode ser atribu?da ? sua a??o citost?tica ou indu??o da apoptose por essas glucanas

Made available in DSpace on 2014-12-17T14:03:38Z (GMT). No. of bitstreams: 1 MoniqueGCFA_DISSERT_PARCIAL.pdf: 1765775 bytes, checksum: 3815c2c3560cb6ce59df27ea29b474ca (MD5) Previous issue date: 2011-07-18 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Marine algae are one of the major sources of biologic compounds. In extracellular matrix of these organisms there are sulfated polysaccharides that functions as structural components and provides protection against dehydration. The fraction 1.0 (F1.0) rich in sulfated galactans obtained from red seaweed Hypnea musciformis was physicochemical characterized and evaluated for pharmacologic activity through antioxidant activity, cytotoxic action on erythrocytes, anticoagulant, stimulatory action under antithrombotic heparan sulfate synthesis and their effects on cell proliferation and cycle cell progression. The main components of F1.0 were carbohydrates (49.70 ? 0.10%) and sulfate (44.59 ? 0.015%), presenting phenolic compounds (4.79 ? 0.016%) and low protein contamination (0.92 ? 0.001%). Fraction 1.0 showed polidisperse profile and signs in infrared analysis in 1262, 1074 and 930, 900 and 850 attributed to sulfate esters S=O bond, presence of a 3,6- anidrogalactose C-O bond, non-sulfated ?-D-galactose and a C-O-SO4 bond in galactose C4, respectively. The fraction rich in sulfated galactans exhibited strong antioxidant action under lipid peroxidation assay with IC50 of 0.003 mg/mL. Besides the inhibition of hemolysis induced by H2O2 in erythrocytes treated with F1.0, this fraction did not promote significant cytotoxity under erythrocytes membranes. F1.0 exhibited low anticoagulant activity causing moderate direct inhibition of enzimatic activity of thrombin. This fraction promoted stimulation around of 4.6 times on this synthesis of heparan sulfate (HS) by rabbit aortic endothelial cells (RAEC) in culture when was compared with non treated cells. The fraction of this algae displayed antiproliferative action under RAEC cells causing incresing on cell number on S fase, blocking the cycle cell progression. Thus F1.0 presented cytostatic and no cytotoxic action under this cell lineage. These results suggest that F1.0 from H. musciformis have antioxidant potential which is a great effect for a compound used as food and in food industry which could be an alternative to food industry to prevent quality decay of lipid containing food due to lipid peroxidation. These polysaccharides prevent the lipid peroxidation once the fraction in study exhibited strong inhibitory action of this process. Furthermore that F1.0 present strong antithrombotic action promoting the stimulation of antithrombotic HS synthesis by endothelial cells, being important for thrombosis preventing, by its inhibitory action under reactive oxygen species (ROS) in some in vitro methods, being involved in promotion of hypercoagulability state. / Algas marinhas s?o uma das principais fontes de compostos biologicamente ativos. Na matriz extracelular desses organismos existem os polissacar?deos sulfatados que funcionam como componente estrutural prevenindo-a contra desidrata??o. A fra??o 1,0 (F1,0) rica em galactanas sulfatadas obtida da alga vermelha Hypnea musciformis foi caracterizada fisicoquimicamente e avaliada quanto a atividade farmacol?gica por meio de ensaios de atividade antioxidante, a??o citot?xica sobre hem?cias, atividade anticoagulante, a??o estimulat?ria sobre a s?ntese de heparam sulfato antitromb?tico e seus efeitos na prolifera??o e progress?o do ciclo celular. Os principais componentes da F1,0 foram carboidratos (49,70 ? 0,10%) e sulfato (44,59 ? 0,015%), apresentando compostos fen?licos (4,79 ? 0,016%) e baixa contamina??o prot?ica (0,92 ? 0,001%). F1,0 mostrou perfil polidisperso e sinais na an?lise de infravermelho em 1262, 1074 e 930, 900 e 850 cm-1 atribu?dos a liga??es S=O de ?steres de sulfato, presen?a de liga??o C-O de 3,6-anidrogalactose, ?-D-galactose n?o sulfatada e liga??o C-O-SO4 no C4 da galactose, respectivamente. A fra??o rica em galactanas sulfatadas exibiu forte a??o antioxidante sobre o ensaio de peroxida??o lip?dica com IC50 de 0,003 mg/mL. Al?m da alta inibi??o da hem?lise induzida por H2O2 em hem?cias humanas tratadas com F1,0, esta fra??o n?o promoveu citotoxicidade significativa sobre a membrana de hem?cias. F1,0 exibiu baixa atividade anticoagulante, causando moderada inibi??o direta da atividade enzim?tica da trombina. Esta fra??o promoveu estimula??o de cerca de 4,6 vezes na s?ntese de heparam sulfato (HS) pelas c?lulas endoteliais da aorta de coelho (RAEC), em cultura, quando comparadas com as c?lulas n?o tratadas com F1,0. A fra??o dessa alga mostrou atividade antiproliferativa sobre as c?lulas RAEC, causando aumento no n?mero de c?lulas na fase S, impedindo a progress?o do ciclo celular. Assim, F1,0 apresentou a??o citost?tica e n?o citot?xica sobre esta linhagem celular. Esses resultados sugerem que F1,0 de H. musciformis tem potencial antioxidante, efeito importante para um composto utilizado como alimento e na ind?stria aliment?cia, podendo ser uma alternativa na ind?stria aliment?cia para a preven??o do decaimento da qualidade dos alimentos contendo lip?dio devido a peroxida??o lip?dica, uma vez que a fra??o em estudo exibiu forte a??o inibit?ria sobre a peroxida??o lip?dica. Al?m disso F1,0 apresenta forte a??o antitromb?tica promovendo a estimula??o da s?ntese de HS antitromb?tico pelas c?lulas endoteliais, sendo ?til na preven??o da trombose, devido tamb?m a sua a??o inibit?ria sobre as esp?cies reativas do oxig?nio (ROS) em alguns sistemas in vitro, estando envolvidos na promo??o de estado de hipercoagulabilidade.

Made available in DSpace on 2014-12-17T14:03:44Z (GMT). No. of bitstreams: 1 ThiagoGC_DISSERT.pdf: 2736665 bytes, checksum: 120a3ba44fefe1ccd9373aeb0ff1629f (MD5) Previous issue date: 2014-02-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Seaweeds are a major source of biologically active compounds . In the extracellular matrix of these organisms are sulfated polysaccharides that functions as structural components preventing it against dehydration. The fraction 0.9 (FucB) rich in sulfated fucans obtained from brown seaweed Dictyota menstrualis was chemical characterized and evaluated for pharmacological activity by testing anticoagulant activity, stimulatory action on the synthesis of an antithrombotic heparan sulfate, antioxidant activity and its effects in cell proliferation. The main components were FucB carbohydrates (49.80 ? 0.10 %) and sulfate (42.30 ? 0.015 %), with phenolic compounds ( 3.86 ? 0.016 %) and low protein contamination ( 0.58 ? 0.001 % ) . FucB showed polydisperse profile and analysis of signals in the infrared at 1262, 1074 and 930 cm -1 and 840 assigned to S = O bonds sulfate esters , CO bond presence of 3,6- anhydrogalactose , &#946; -D- galactose non- sulfated sulfate and the axial position of fucose C4 , respectively. FucB exhibited moderate anticoagulant activity , the polysaccharides prolonged time (aPTT ) 200 ug ( > 90s ) partial thromboplastin FucB no effect on prothrombin time (PT), which corresponds to the extrinsic pathway of coagulation was observed. This stimulation promoted fraction of about 3.6 times the synthesis of heparan sulfate (HS) by endothelial cells of the rabbit aorta ( RAEC ) in culture compared with cells not treated with FucB . This has also been shown to compete for the binding site with heparin. The rich fraction sulfated fucans exhibited strong antioxidant activity assays on total antioxidant (109.7 and 89.5 % compared with BHT and ascorbic acid standards ) , reducing power ( 71 % compared to ascorbic acid ) and ferric chelation ( 71 , comparing with 5 % ascorbic acid). The fraction of algae showed cytostatic activity on the RAEC cells revealed that the increase of the synthesis of heparan sulfate is not related to proliferation. FucB showed antiproliferative action on cell lines modified as Hela and Hep G2 by MTT assay . These results suggest that FucB Dictyota menstrualis have anticoagulant , antithrombotic , antioxidant potential as well as a possible antitumor action, promoting the stimulation of the synthesis of antithrombotic HS by endothelial cells and is useful in the prevention of thrombosis, also due to its inhibitory action on species reactive oxygen ( ROS ) in some in vitro systems , being involved in promoting a hypercoagulable state / Algas marinhas s?o uma das principais fontes de compostos biologicamente ativos. Na matriz extracelular desses organismos existem os polissacar?deos sulfatados que funcionam como componente estrutural prevenindo-a contra desidrata??o. A fra??o 0,9 (FucB) rica em fucanas sulfatadas obtida da alga marrom Dictyota menstrualis foi caracterizada quimicamente e avaliada quanto a atividade farmacol?gica por meio de ensaios de atividade anticoagulante, a??o estimulat?ria sobre a s?ntese de heparam sulfato antitromb?tico, atividade antioxidante e seus efeitos na prolifera??o celular. Os principais componentes da FucB foram carboidratos (49,80 ? 0,10%) e sulfato (42,30 ? 0,015%), apresentando compostos fen?licos (3,86 ? 0,016%) e baixa contamina??o prot?ica (0,58 ? 0,001%). FucB mostrou perfil polidisperso e sinais na an?lise de infravermelho em 1262, 1074 e 930 e 840 cm-1 atribu?dos a liga??es S=O de ?steres de sulfato, presen?a de liga??o C-O de 3,6-anidrogalactose, &#946;-D-galactose n?o sulfatada e sulfato na posi??o axial do C4 da fucose, respectivamente. FucB exibiu moderada atividade anticoagulante, este polissacar?deo prolongou o tempo de tromboplastina parcial activada (aPTT) a 200 ug (>90s) n?o foi observado qualquer efeito de FucB sobre o tempo de protrombina (PT), que corresponde a via extr?nseca da coagula??o. Esta fra??o promoveu estimula??o cerca de 3,6 vezes na s?ntese de heparam sulfato (HS) pelas c?lulas endoteliais da aorta de coelho (RAEC), em cultura, quando comparadas com as c?lulas n?o tratadas com FucB. Esta tamb?m demonstrou competir pelo s?tio de liga??o com a heparina. A fra??o rica em fucanas sulfatadas exibiu forte a??o antioxidante sobre os ensaios de antioxidante total (109,7 e 89,5% comparados com padr?es BHT e ?cido asc?rbico), poder redutor (71% comparado ao ?cido asc?rbico) e quela??o f?rrica (71,5% comparando com ?cido asc?rbico). A fra??o dessa alga mostrou atividade citost?tica sobre as c?lulas RAEC revelando que o aumento da s?ntese de heparan sulfato n?o est? relacionado ? prolifera??o. FucB apresentou a??o antiproliferativa sobre linhagens celulares modificadas como Hela e Hep G2 pelo ensaio de MTT. Esses resultados sugerem que FucB de Dictyota menstrualis tem potencial anticoagulante, antitromb?tico, antioxidante bem como uma poss?vel a??o antitumoral, promovendo a estimula??o da s?ntese de HS antitromb?tico pelas c?lulas endoteliais, sendo ?til na preven??o da trombose, devido tamb?m a sua a??o inibit?ria sobre as esp?cies reativas do oxig?nio (ROS) em alguns sistemas in vitro, estando envolvidos na promo??o de estado de hipercoagulabilidade