Made available in DSpace on 2014-12-17T14:03:42Z (GMT). No. of bitstreams: 1 LucianaFC_DISSERT.pdf: 1219461 bytes, checksum: 86df41deecc523bb278b7a9bc3b86ec9 (MD5) Previous issue date: 2013-03-04 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / In recent years the heparin has been the subject of several studies that aim to expand its use as a therapeutic agent, due to its ability to modulate the activity of various proteins that play important roles in the regulation of pathophysiological processes. In several experiments and preclinical trials, heparin has demonstrated an anti-inflammatory role. However, its clinical use is limited, due to its strong anticoagulant activity and hemorrhagic complications. For this reason, considerable efforts have been employed in discovery of heparin analogous (heparinoid) with reduced side effects, that retain the anti-inflammatory properties of heparin. In this context, a heparinoid obtained from the head of Litopenaeus vannamei shrimp, which presents a structural similarity to heparin, showed, in previous studies, anti-inflammatory activity in a model of acute peritonitis with reduced anticoagulant effect in vitro and low hemorrhagic activity. Thus, the present work had as objective to evaluate the effect the heparinoid of the cephalothorax of gray shrimp on the acute inflammatory response in different times (3 or 6 hours after the induction of inflammatory stimulus), using the model of acute peritonitis induced in mice. It was also analyzed the HL effect over the activity of elastase, an enzyme involved in leukocyte recruitment. Furthermore to check if the different doses of heparin and heparinoid change the hemostatic balance in vivo, was assessed the effect of these compounds on the plasma clotting time in animals submitted to inflammation. The results show that in 3 hours, all doses of heparinoid were able to prevent efficiently in the acute inflammatory process without any anticoagulant effects, unlike the extrapolation dose of heparin, which has induced a large hemorrhage due its high anticoagulant activity. However, 6 hours after induction of inflammation, only the dosages of 0.1 and 1.0 μg/Kg of heparin and 1.0 μg/Kg of heparinoid kept anti-migratory effect, without changing of the hemostatic balance. These results indicate that the anti-migratory effect of theses compounds depends on the dosage and time of inflammatory stimulus. The HL and heparin were also able to inhibit the activity of the enzyme elastase. The discovery of this bioactive compound in the cephalothorax of shrimps can arouse great interest in biotechnology, since this compound could be useful as a structural model interesting for the development of new therapeutic agents for peritonitis / Nos ?ltimos anos a heparina tem sido alvo de diversos estudos que visam ampliar seu uso como agente terap?utico, devido ? sua habilidade de modular a atividade de v?rias prote?nas que desempenham pap?is importantes na regula??o de processos fisiopatol?gicos. Em diversos experimentos e ensaios pr?-cl?nicos, a heparina tem demonstrado papel anti-inflamat?rio. Entretanto, seu uso cl?nico ? limitado, devido ? sua forte atividade anticoagulante e complica??es hemorr?gicas. Por essa raz?o, consider?vel esfor?o tem sido empregado na descoberta de an?logos da heparina (heparin?ide) com reduzidos efeitos colaterais, que retenham as propriedades anti-inflamat?rias da heparina. Nesse contexto, um heparin?ide (HL) obtido da cabe?a do camar?o Litopenaeus vannamei, de semelhan?a estrutural ? heparina, apresentou em estudos pr?vios atividade anti-inflamat?ria em modelo de peritonite aguda, com reduzido efeito anticoagulante in vitro e baixa atividade hemorr?gica. Assim, o presente trabalho teve como objetivo avaliar o efeito deste heparin?ide sobre a resposta inflamat?ria aguda em diferentes tempos (3 ou 6 horas ap?s a indu??o do est?mulo inflamat?rio), utilizando o modelo de peritonite aguda induzida em camundongos. Foi analisado tamb?m o efeito do HL sobre a atividade da elastase, uma enzima envolvida no recrutamento de leuc?citos. Al?m disso, para verificar se as diferentes doses do heparin?ide e da heparina alteram o equil?brio hemost?tico in vivo, foi avaliado o efeito desses compostos sobre o tempo de coagula??o do plasma nos animais submetidos ? inflama??o. Os resultados revelam que em 3 horas, todas as doses do heparin?ide foram capazes de interferir de forma eficiente no processo inflamat?rio agudo sem apresentar efeito anticoagulante e interfer?ncia no equil?brio hemost?tico, ao contr?rio da dose de extrapola??o da heparina que induziu forte hemorragia, al?m de apresentar alta atividade anticoagulante. Entretanto, no tempo de 6 horas ap?s a indu??o da inflama??o, apenas as doses de 0,1 e 1,0 μg/Kg da heparina e 1,0 μg/Kg do heparin?ide mantiveram o efeito antimigrat?rio, sem alterar o equil?brio hemost?tico. Esses resultados indicam que o efeito antimigrat?rio depende do tempo e da dose administrada. O HL e a heparina tamb?m foram capazes de inibir a atividade da enzima elastase. A descoberta desse composto bioativo no cefalot?rax do camar?o poder? despertar grande interesse biotecnol?gico, pois este composto poderia servir como um modelo estrutural interessante para o desenvolvimento de novos agentes terap?uticos espec?ficos para a peritonite

Made available in DSpace on 2014-12-17T14:03:41Z (GMT). No. of bitstreams: 1 VanessaOS.pdf: 1126102 bytes, checksum: dba464c890585c6be32a3fd2dc600a81 (MD5) Previous issue date: 2006-06-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Sulfated Polysaccharides with unique chemical structures and important biological activities has been found in a diversity of sea invertebrates. For that, to exist a huger interest on the biotechnology field in the research theses sulfated compounds isolated from sea organisms. Despite the privileged brazilian position for these compounds attainment, there are still a few scientific informations about the isolated substances and their biological activities. A head the displayed, the present work has for objectives, to evaluate the pharmacological properties of the glycosaminoglycans isolated from the sea shrimp Litopenaeus schimitti on homeostasis, blood coagulation, leukocytes migration and platelet/leukocyte adhesion. For this, yhe glycosaminoglycans were extracted from crustacean tissues by proteolysis, fractionation with acetone and later submitted to pharmacological assays. The crustacean tissues showed compounds heparin-like, with anticoagulant activity of 45 IU/mg and 90 IU/mg, respectively. These molecules showed low residual hemorrhagic effects in the tested concentration (100 ?g/mL), when compared to unfractionated commercial heparin (UFH). Another dermatan sulfate-like compound, predominately constituted for disulfated disaccharides, was isolated from crustacean abdomen. This compound showed an efficient effect on leukocytes migration inhibition, in the concentration of 15 ?g/mL, reducing the cellular infiltration in 65% when compared to the controlled animals. In this same concentration, the DS reduced in 60% the protein concentration of the peritoneal exudates. In the concentration, this compound of 0.5 mg/mL, it was capable to reduce in 40% platelet/leukocytes adhesion. Our data demonstrate that these sulfated polysaccharides isolated from the shrimp L. schimitti will can be used as bioactive compounds, appearing as active principles for pharmacological development, anticoagulants and inflammatory response regulators / Polissacar?deos sulfatados com caracter?sticas estruturais distintas e importantes atividades biol?gicas t?m sido encontrados em uma diversidade de invertebrados marinhos. Por isso existe um grande interesse no campo da biotecnologia na pesquisa destes compostos sulfatados isolados de organismos aqu?ticos. No entanto, apesar da posi??o privilegiada do Brasil para a obten??o destes compostos, ainda s?o poucas as informa??es cient?ficas sobre as subst?ncias isoladas e suas atividades biol?gicas. Diante do exposto, este presente trabalho teve por objetivos avaliar os potenciais farmacol?gicos dos glicosaminoglicanos (GAGs) isolados do camar?o marinho Litopenaeus schimitti, sobre a hemostasia, coagula??o sang??nea, migra??o leucocit?ria e ades?o celular. Para isso os GAGs foram extra?dos dos tecidos do crust?ceo mediante prote?lise, fracionamento com acetona e posteriormente submetidos aos ensaios farmacol?gicos. Os tecidos do crust?ceo, abd?men e cefalot?rax, apresentaram compostos semelhantes ? heparina (heparin?ides) com atividade anticoagulante de 45 UI/mg e 90 UI/mg, respectivamente. Estas mol?culas apresentaram baixo efeito hemorr?gico residual na concentra??o de 100 ?g/mL, quando comparada com a heparina comercial n?o fracionada (HNF). Um outro composto semelhante ao dermatam sulfato (DS), constitu?do predominantemente por dissacar?deos dissulfatados foi isolado do abd?men do crust?ceo. Este composto apresentou, na concentra??o de 15 ?g/?L, uma inibi??o significativa (P<0.01) da migra??o leucocit?ria, reduzindo a infiltra??o celular em 65% quando comparado com os animais controle. Nessa mesma concentra??o o DS reduziu em 60% a concentra??o de prote?nas do lavado peritonial. As an?lises qualitativas da composi??o celular do exudato peritonial foram similares ao encontrado para os animais controles em todas as concentra??es testadas. Na concentra??o de 0,5 mg/mL foi capaz de reduzir em 40% a ades?o das plaquetas aos leuc?citos. Os dados obtidos demonstram que estes polissacar?deos sulfatados isolados do camar?o L.schimitti podem vir a ser utilizados como compostos bioativos, podendo surgir como princ?pios ativos para o desenvolvimento de f?rmacos, anticoagulantes e moduladores da resposta inflamat?ria