Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2015-10-20T15:14:03Z No. of bitstreams: 1 JulianaFelixDaSilva_DISSERT.pdf: 3172083 bytes, checksum: 0b4b23addc49fd9790ec430dc1ac0b32 (MD5) / Approved for entry into archive by clediane guedes (clediane@bczm.ufrn.br) on 2015-10-20T15:38:30Z (GMT) No. of bitstreams: 1 JulianaFelixDaSilva_DISSERT.pdf: 3172083 bytes, checksum: 0b4b23addc49fd9790ec430dc1ac0b32 (MD5) / Made available in DSpace on 2015-10-20T15:38:31Z (GMT). No. of bitstreams: 1 JulianaFelixDaSilva_DISSERT.pdf: 3172083 bytes, checksum: 0b4b23addc49fd9790ec430dc1ac0b32 (MD5) Previous issue date: 2014-02-25 / Os acidentes of?dicos s?o um grave problema de sa?de p?blica. Cerca de 90 % dos casos na Am?rica Latina s?o associados a serpentes do g?nero Bothrops. Atualmente, a principal forma de tratamento dispon?vel ? a soroterapia antiveneno, que apresenta algumas desvantagens, tais como inefici?ncia quanto a efeitos locais, riscos de rea??es imunol?gicas, custo elevado e dif?cil acesso em algumas regi?es. Nesse contexto, a busca por novas alternativas para o tratamento de picadas de serpentes se faz relevante. Jatropha gossypiifolia L., esp?cie vegetal popularmente conhecida no Brasil como ?pinh?o-roxo?, ? bastante utilizada na medicina popular como antiof?dica. Portanto, o objetivo do presente estudo ? avaliar a atividade antiof?dica desta esp?cie frente ?s atividades enzim?ticas e biol?gicas do veneno da serpente Bothrops jararaca. O extrato aquoso das folhas foi preparado por decoc??o. Os estudos de inibi??o foram realizados in vitro, pr?-incubando-se quantidade fixa de veneno com diferentes quantidades do extrato por 60 min a 37?C, e in vivo, atrav?s do tratamento oral ou intraperitoneal dos animais, em diferentes doses, 60 min antes da inje??o do veneno. A atividade proteol?tica sobre a azocase?na foi eficientemente inibida, o que indica a??o inibit?ria frente metaloproteases (SVMPs) e/ou serinoproteases (SVSPs). O extrato inibiu a atividade fibrinogenol?tica, o que tamb?m foi confirmado por meio de zimografia, onde foi poss?vel visualizar que o extrato inibe preferencialmente enzimas fibrinogenol?ticas de 26 e 28 kDa. As atividades coagulante sobre o fibrinog?nio e sobre o plasma foram significativamente inibidas, sugerindo uma a??o inibit?ria sobre enzimas semelhantes ? trombina (SVTLEs), bem como sobre toxinas ativadoras de fatores da coagula??o. O extrato prolongou o tempo de tromboplastina parcial ativada (TTPa), o que sugere que o extrato inibe tanto SVTLEs quanto a trombina end?gena. A atividade defibrinogenante in vivo foi inibida eficientemente pelo extrato pela via oral, confirmando os resultados anteriores. A hemorragia local tamb?m foi inibida significativamente pela via oral, indicando uma a??o inibit?ria frente SVMPs hemorr?gicas. A atividade fosfolip?sica n?o foi inibida. Mesmo assim, as atividades edematog?nica e miot?xica foram eficientemente inibidas, por via oral e intraperitoneal, o que pode indicar a??o inibit?ria do extrato frente fosfolipases (PLA2) do tipo Lys49 e/ou SVMPs, ou ainda uma a??o anti-inflamat?ria contra os mediadores qu?micos end?genos. Com rela??o ao poss?vel mecanismo de a??o, observou-se que o extrato n?o apresenta atividade proteol?tica, por?m apresenta capacidade de precipitar prote?nas. Adicionalmente, o extrato apresentou significativa atividade antioxidante em diferentes modelos, o que pode justificar ao menos parcialmente a atividade antiof?dica apresentada. A atividade quelante de metais apresentada pelo extrato pode ser correlacionada ? inibi??o de SVMPs, uma vez que estas s?o enzimas dependentes de metais. A an?lise fitoqu?mica revelou a presen?a de a??cares, alcaloides, flavonoides, taninos, terpenos e/ou esteroides e prote?nas, dos quais os flavonoides podem ser apontados como compostos majorit?rios, tendo em vista o perfil cromatogr?fico obtido por cromatografia em camada delgada (CCD). Em conclus?o, os resultados demonstram que o decocto das folhas de J. gossypiifolia apresenta potencial atividade antiof?dica, inclusive atuando sobre os efeitos locais da picada, sugerindo que esta esp?cie pode ser utilizada como uma nova fonte de mol?culas bioativas contra venenos botr?picos. / Antiophidic activity from decoct of Jatropha gossypiifolia L. leaves against Bothrops jararaca venom. Snakebites are a serious worldwide public health problem. In Latin America, about 90 % of accidents are attributed to snakes from Bothrops genus. Currently, the main available treatment is the antivenom serum therapy, which has some disadvantages such as inability to neutralize local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies to treat snakebites is relevant. Jatropha gossypiifolia L., a medicinal plant popularly known in Brazil as ?pinh?o-roxo?, is very used in folk medicine as antiophidic. So, the aim of this study is to evaluate the antiophidic properties of this species against enzymatic and biological activities from Bothrops jararaca snake venom. The aqueous leaf extract of J. gossypiifolia was prepared by decoction. The inhibition studies were performed in vitro, by pre-incubation of a fixed amount of venom with different amounts of extract from J. gossypiifolia for 60 min at 37 ?C, and in vivo, through oral or intraperitoneal treatment of animals, in different doses, 60 min before venom injection. The proteolytic activity upon azocasein was efficiently inhibited, indicating inhibitory action upon metalloproteinases (SVMPs) and/or serine proteases (SVSPs). The extract inhibited the fibrinogenolytic activity, which was also confirmed by zymography, where it was possible to observe that the extract preferentially inhibits fibrinogenolytic enzymes of 26 and 28 kDa. The coagulant activity upon fibrinogen and plasma were significantly inhibited, suggesting an inhibitory action upon thrombin-like enzymes (SVTLEs), as well as upon clotting factor activators toxins. The extract prolonged the activated partial thromboplastin time (aPTT), suggesting an inhibitory action toward not only to SVTLEs, but also against endogenous thrombin. The defibrinogenating activity in vivo was efficiently inhibited by the extract on oral route, confirming the previous results. The local hemorrhagic activity was also significantly inhibited by oral route, indicating an inhibitory action upon SVMPs. The phospholipase activity in vitro was not inhibited. Nevertheless, the edematogenic and myotoxic activities were efficiently inhibited, by oral and intraperitoneal route, which may indicate an inhibitory effect of the extract upon Lys49 phospholipase (PLA2) and/ or SVMPs, or also an anti-inflammatory action against endogenous chemical mediators. Regarding the possible action mechanism, was observed that the extract did not presented proteolytic activity, however, presented protein precipitating action. In addition, the extract showed significant antioxidant activity in different models, which could justify, at least partially, the antiophidic activity presented. The metal chelating action presented by extract could be correlated with SVMPs inhibition, once these enzymes are metal-dependent. The phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins, from which the flavonoids could be pointed as major compounds, based on chromatographic profile obtained by thin layer chromatography (TLC). In conclusion, the results demonstrate that the J. gossypiifolia leaves decoct present potential antiophidic activity, including action upon snakebite local effects, suggesting that this species may be used as a new source of bioactive molecules against bothropic venom.