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.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-07-25T20:53:39Z No. of bitstreams: 1 JacyraAntunesDosSantosGomes_DISSERT.pdf: 2709008 bytes, checksum: b8536d693075ca072ab23c36cf351557 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-08-02T20:59:13Z (GMT) No. of bitstreams: 1 JacyraAntunesDosSantosGomes_DISSERT.pdf: 2709008 bytes, checksum: b8536d693075ca072ab23c36cf351557 (MD5) / Made available in DSpace on 2016-08-02T20:59:13Z (GMT). No. of bitstreams: 1 JacyraAntunesDosSantosGomes_DISSERT.pdf: 2709008 bytes, checksum: b8536d693075ca072ab23c36cf351557 (MD5) Previous issue date: 2015-03-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Os acidentes of?dicos representam um s?rio problema em sa?de p?blica nos pa?ses tropicais e subtropicais, sendo o g?nero Bothrops o maior respons?vel pelos acidentes no Brasil e em toda a Am?rica Latina (correspondendo cerca de 90 % dos casos). Os efeitos locais (dor, edema, hemorragia e necrose tecidual) e sist?micos (altera??es cardiovasculares, choque e dist?rbios da coagula??o sangu?nea) causados pela pe?onha das serpentes do g?nero Bothrops s?o devido aos in?meros componentes prot?icos e n?o-prot?icos (carboidratos, lip?dios, nucleot?deos, aminas biog?nicas e v?rios ?ons), que fazem parte da constitui??o da pe?onha. A ?nica forma de terapia cientificamente validada ? a soroterapia antiveneno, que, no entanto, n?o ? eficaz com rela??o aos efeitos locais produzidos e, al?m disso, pode ocasionar s?rias rea??es de hipersensibilidade aos pacientes. Sendo assim, a busca por novas alternativas ? soroterapia se faz importante, e nesse contexto, muitas plantas medicinais v?m se destacando pelo uso popular como antiof?dicas. Dentre essas plantas, pode-se citar a esp?cie Jatropha mollissima (Euphorbiaceae) que apresenta amplo uso popular na medicina tradicional como antiof?dica, anti-inflamat?ria, antimicrobiana e antit?rmica. Portanto, esse trabalho tem como objetivo a avalia??o do potencial neutralizante dos efeitos locais induzidos pelas pe?onhas de Bothrops erythromelas e Bothrops jararaca pelo extrato aquoso das folhas de J. mollissima. O extrato das folhas foi preparado por decoc??o, fracionado (por meio de parti??o l?quido-l?quido) e caracterizado por cromatografia em camada delgada (CCD) e Cromatografia L?quida de Alta Efici?ncia (CLAE). A atividade antiof?dica do extrato foi avaliada nos modelos de edema de pata, peritonite, hemorragia e miotoxicidade induzidos pelas pe?onhas de B. erythromelas e B. jararaca sendo utilizados camundongos Swiss de ambos os sexos. Em todos os modelos o extrato foi avaliado pela via intraperitoneal nas doses de 50, 100 e 200 mg/kg, sendo administrado 30 minutos antes da inje??o da pe?onha (protocolo de pr?-tratamento). Manchas sugestivas da presen?a dos flavon?ides: apigenina, luteolina, orientina, isoorientina, vitexina e vitexina-2-O-ramnos?deo foram detectadas no extrato atrav?s da co-CCD. Por meio de CLAE foram identificados os flavon?ides isoorientina, orientina, vitexina e isovitexina. Todas as doses testadas do extrato de J. mollissima reduziram o edema de pata induzido pelas pe?onhas com intensidade similar ? dexametasona. O extrato aquoso das folhas de J. mollissima, em todas as doses avaliadas, inibiu a migra??o celular induzida por B. erythromelas e B. jararaca promovendo a inibi??o do recrutamento tanto de c?lulas mononucleares quanto das c?lulas polimorfonucleares. A hemorragia local induzida pela pe?onha de B. jararaca foi inibida significativamente pelo extrato. Ambas as pe?onhas foram inibidas pelo extrato na atividade miot?xica. Esses resultados indicam que o extrato aquoso das folhas de J. mollissima apresenta potencial propriedade antiof?dica sobretudo com rela??o ao efeitos locais, o que poderia justificar o uso dessa planta na medicina tradicional e na terapia complementar como antiof?dica. / Snakebites are a serious public health problem in tropical and subtropical countries and Bothrops genus is responsible for the accidents in Brazil and throughout Latin America (90% of cases). The local effects (pain, edema, hemorrhage and myonecrosis) and systemic (cardiovascular alterations, shock and blood clotting disorders) caused by the venom of Bothrops are due to the numerous protein and non-protein components, which are part of the constitution of the poison. The only form of therapy is scientifically validated antivenom serum therapy which, however, is not effective with respect to local effects produced, risk of immunological reactions, high cost and difficult access in some regions. Thus, the search for new alternatives to serum therapy becomes important, and in this context, many medicinal plants have been highlighted by the popular use as antiophidic. Among these plants, we can mention the species Jatropha mollissima (Euphorbiaceae) which has popular use in traditional medicine as antiophidic, anti-inflammatory, antimicrobial and antipyretic. Therefore, this study aims to evaluate the neutralizing potential of local effects induced by the venom of Bothrops erythromelas and Bothrops jararaca with the aqueous extract of the leaves of J. mollissima. The leaf extracts were prepared by decoction, fractionated (by liquid-liquid partition) and characterized by thin layer chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Antiophidic activity of the extract was evaluated in model of paw edema, peritonitis, bleeding and myotoxicity induced by venoms of B. jararaca and B. erythromelas. In all models, the extract was evaluated by intraperitoneal route at the doses of 50, 100 and 200 mg/kg, administered 30 minutes prior to injection of the venom (pretreatment protocol). Stains suggestive of the presence of flavonoids: apigenin, luteolin, orientin, isoorientin, vitexin and vitexin-2-O-rhamnoside were detected in the extract by co-CCD. By means of HPLC were identified isoorientin, orientin, vitexin and isovitexin. All tested doses of J. mollissima extract reduced the paw edema induced by the venom with intensity similar to dexamethasone. The aqueous extract of J. mollissima leaves on all evaluated doses, inhibited cell migration induced by B. jararaca and B. erythromelas promoting inhibition of recruitment of mononuclear cells and the polymorphonuclear cells. Local bleeding induced by B. jararaca venom was significantly inhibited by the extract. Both venoms were inhibited by the extract in myotoxic activity. These results indicate that the aqueous extract of J. mollissima leaves have snakebite potential, particularly with respect to local effects, which may justify the use of this plant in traditional medicine and complementary therapy as anti-venom serum.