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Efeito citot?xico e potencial anti-inflamat?rio de Pseudobrickellia brasiliensis (Spreng) R, M. King & H. Rob (arnica-do-campo) em c?lulas mononucleares do sangue perif?rico humano, in vitro.Almeida, Valeria Gomes de January 2012 (has links)
?rea de concentra??o: Farmacologia de produtos naturais e plantas medicinais. / Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-10-23T16:44:04Z
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Previous issue date: 2012 / A Pseudobrickellia brasiliensis (Asteraceae), conhecida como arnica-do-campo, ? uma planta nativa da flora brasileira cuja prepara??o alco?lica tem sido utilizada como anti-inflamat?rio pela medicina popular. No entanto, os mecanismos biol?gicos envolvidos nessa atividade, ainda n?o foram devidamente elucidados. Nesse contexto, o presente estudo avaliou o potencial anti-inflamat?rio de P. brasiliensis atrav?s da an?lise da produ??o de citocinas anti e pr?-inflamat?rias por leuc?citos mononucleares do sangue perif?rico. Foram utilizados extratos de P. brasiliensis ap?s extra??o com os solventes Acetato de Etila (ACE) e Etanol (ETA), ambos dilu?dos em Dimetilsulf?xido (DMSO), bem como o extrato aquoso dilu?do em PBS (AQU). A citotoxicidade dos extratos, ap?s 24h ou 5 dias de cultura celular, foi avaliada por meio da t?cnica de exclus?o com Azul de Tripan e, tais resultados foram confirmados por meio de ensaios de apoptose e necrose, ap?s marca??o com Anexina V-FITC e Iodeto de Prop?deo. No ensaio de inibi??o da produ??o de citocinas, C?lulas Mononucleares do Sangue Perif?rico humano (PBMC) foram incubadas em meio de cultura, contendo Miristato-Acetato de Forbol (PMA) e Ionomicina como agentes estimuladores, na aus?ncia ou presen?a do extrato AQU nas concentra??es finais de 0,025 mg/mL, 0,05 mg/mL ou 0,1 mg/mL. Tamb?m, foi avaliado o efeito do extrato AQU sobre a resposta proliferativa de linf?citos ao est?mulo mitog?nico Fitohemaglutinina (PHA) como teste complementar na an?lise de ativa??o celular. De acordo com os dados obtidos, os extratos ACE e ETA apresentaram um alto grau de citotoxicidade, sendo, portanto exclu?dos das an?lises posteriores, que avaliaram o perfil de produ??o de citocinas e prolifera??o. O extrato AQU, al?m de n?o induzir apoptose e necrose, n?o demonstrou a??o t?xica, mesmo ap?s 5 dias de cultura. Nas culturas tratadas com AQU na concentra??o final de 0,1 mg/mL foi observada redu??o significativa da express?o de IFN-? e TNF-? em resposta ? estimula??o com PMA e ionomicina. O efeito inibit?rio do extrato AQU, tamb?m, pode ser observado por meio da diminui??o da resposta proliferativa de linf?citos ao PHA. Estes achados sugerem que o efeito anti-inflamat?rio atribu?do ? P.brasiliensis na medicina popular pode, pelo menos em parte, ser devido ? inibi??o da produ??o de citocinas pr?-inflamat?rias por linf?citos, bem como da resposta proliferativa. Ensaios futuros, utilizando subst?ncias isoladas e identificadas a partir do extrato aquoso, s?o necess?rios para fortalecer os dados aqui apresentados e para demonstrar quais os mecanismos moleculares envolvidos nas a??es inibit?rias apresentadas no presente trabalho. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, [2012]. / ABSTRACT
Pseudobrickellia brasiliensis (Asteraceae), also known as ?Arnica-do-Campo? is a native plant from Brazilian flora and its alcoholic preparation has been used as anti-inflammatory in folk medicine. However, the biological mechanisms involved in the anti-inflammatory activity are not well elucidated. In this context, the present study evaluated the anti-inflammatory potential of P. brasiliensis by performing an anti- and pro-inflammatory cytokine analysis on human peripheral blood leucocytes as well as cell activation, measured by proliferation assay. We used P. brasiliensis extracts utilizing different solvents including Ethyl Acetate (ACE) and ethanol (ETA), both diluted in dimethylsulfoxide (DMSO), and aqueous extract (AQU) diluted in PBS. After 24h or 5 days, in vitro cytotoxicity promoted by those extracts was evaluated using the exclusion technique with Trypan Blue. Furthermore, apoptosis and necrosis was measured by Annexin V-FITC and propidium iodide staining, after 4 hours of culture. For cytokine production inhibitory assay, humans Peripheral Blood Mononuclear Cells (PBMC) were incubated in medium containing Phorbol Myristate Acetate (PMA) plus ionomycin as stimulating agents in the absence or presence of AQU extract at the final concentrations 0.025 mg/ml, 0.05 mg/ml or 0.1 mg/ml. We also assessed, as a complementary test for cellular activation analysis, the effect of AQU extract on lymphocytes proliferative response after mitogenic stimulation with Phytohemagglutinin (PHA). According to the obtained data, ACE and ETA showed a high cytotoxicity rate and, therefore, they were not used in cytokine and proliferative analysis. The AQU extracts, besides not inducing apoptosis and necrosis, showed no toxic effects even after 5 days of culture. In cultures treated with AQU extract at final concentration of 0.1 mg / ml was observed a significant reduction in IFN-? and TNF-? expression in response to PMA stimulation. The AQU extract inhibitory effect can also be observed by the decrease of lymphocytes proliferative rate in response to PHA stimulation. These findings suggest that P.brasiliensis anti-inflammatory effect can be, at least in part, due PBMC pro-inflammatory cytokines inhibition as well as the proliferative response. Further assays, using aqueous extract isolated and identified substances are needed to strengthen the presented data and to demonstrate the molecular mechanisms involved in inhibitory actions reported in this study.
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Um dermatam sulfato antitromb?tico do camar?o Litopenaeus vanammei inibe a inflama??o e angiog?nesePalhares, Lais Cristina Gusm?o Ferreira 29 March 2016 (has links)
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Previous issue date: 2016-03-29 / A inflama??o ? composta de uma rea??o vascular e outra celular, conferindo diferentes rea??es de tecidos e c?lulas, tanto do ambiente intravascular, como o do ambiente extravascular. ? medida que o processo inflamat?rio ocorre, proteases da coagula??o, em especial a trombina (FIIa), s?o capazes de desencadear diversas respostas celulares na biologia vascular e por isso, frequentemente ? observada a ativa??o de outros sistemas biol?gicos, levando ? complica??es durante um evento inflamat?rio, como a trombose e a angiog?nese. Assim, mol?culas antagonistas desses eventos s?o modelos interessantes para o desenvolvimento de novos f?rmacos anti-inflamat?rios. Neste contexto, destacam-se os glicosaminoglicanos (GAGs), os quais interagem com diversas prote?nas envolvidas em processos biol?gicos importantes, incluindo inflama??o e coagula??o. Por essa raz?o, o presente trabalho teve por objetivo avaliar os potenciais anti-inflamat?rios, antitromb?tico, antiangiog?nico, bem como anticoagulante de GAGs do tipo dermatam sulfato (DS) extra?dos do cefalot?rax do camar?o Litopenaeus vannamei. O composto foi obtido ap?s prote?lise e purifica??o por cromatografia de troca-i?nica. Ap?s total digest?o por liases que digerem compostos tipo DS (condroitinase ABC), sua natureza do tipo DS foi revelada, sendo ent?o denominado DSL. O composto do camar?o mostrou reduzido efeito anticoagulante pelo ensaio de TTPa, por?m apresentou alta atividade anti-IIa, diretamente e via Cofator II da heparina. Sobre a inflama??o, o composto apresentou significativo efeito inibit?rio com redu??o de citocinas pr?-inflamat?rias. Potenciais inibit?rios foram relatados no ensaio antitromb?tico e antiangiog?nico, sendo este ?ltimo dose-dependente. Quanto ? atividade anti-hemost?tica, o polissacar?deo n?o induziu efeito hemorr?gico significativo. Assim, os resultados exibidos pelo composto tipo DS isolado do camar?o, apontam este glicosaminoglicano como alvo biotecnol?gico com perspectivas para o desenvolvimento de novas drogas multipotentes. / Inflammation is combined of a vascular and a cellular reaction, resulting in different cells and tissue responses, both the intravascular and extravascular environment. As the inflammatory process occurs, coagulation proteases, in particular thrombin (FIIa), are able to initiate various cellular responses in vascular biology and therefore is often observed activation of other biological systems, leading to complications during an event inflammatory, such as thrombosis and angiogenesis. Thus, antagonists molecules of these events are interesting models for the development of novel anti-inflammatory drugs. Thereby, it is worth stressing the glycosaminoglycans (GAGs), which are able to interact with several proteins involved in important biological processes, including inflammation and coagulation. Therefore, this study aimed to evaluate the anti-inflammatory, antithrombotic and anti-angiogenic potentials, as well anticoagulant of a dermatan sulfate-like GAG (DS) extracted from the Litopenaeus vannamei cephalotorax. The compound was obtained after proteolysis and purification by ion-exchange chromatography. After total digestion by DS-like compounds digesting lyases (chondroitinase ABC), the DS-like nature was revealed, and then called DSL. The shrimp compound showed reduced anticoagulant effect by the aPTT assay, but high anti-IIa activity, directly and through heparin cofactor II. On inflammation, the compound had a significant inhibitory effect with the reduction of proinflammatory cytokines. Potential Inhibitory were reported in the antithrombotic and anti-angiogenic assay, the latter being dose dependent. As for anti-hemostatic activity, the polysaccharides did not induced significant bleeding effect. Thus, the results shown by the shrimp DS-like compound indicate this glycosaminoglycan as a biotechnology target with prospects for the development of new multipotent drugs.
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An?lise da composi??o qu?mica, atividade citot?xica e inibi??o de citocinas in vitro de prepara??es de partes a?reas da planta ageratum fastigiatumFreitas, Beth?nia Alves de Avelar 13 September 2013 (has links)
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Previous issue date: 2013 / Ageratum fastigiatum ? uma planta utilizada na medicina popular como anti-inflamat?rio e analg?sico, no entanto, poucos estudos foram realizados a fim de detalhar os mecanismos envolvidos nessa atividade. O objetivo deste estudo foi avaliar a atividade anti-inflamat?ria in vitro do ?leo essencial e do extrato dicloromet?nico de A. fastigiatum. Pela t?cnica de exclus?o do azul de tripam por citometria de fluxo foram determinadas concentra??es n?o t?xicas das prepara??es de A. fastigiatum. As concentra??es n?o t?xicas do ?leo essencial foram 5x10-3 e 1x10-2 ?L/mL. Essas concentra??es foram utilizadas para a pesquisa do potencial anti-inflamat?rio do ?leo essencial, medido por meio da an?lise do perfil de citocinas pro (TNF- ? e IFN- ?) e anti-inflamat?rias (IL-10), em culturas de leuc?citos humanos estimulados e n?o estimulados com PMA (acetato de forbol miristato) . Os dados demonstraram que ambas as concentra??es inibiram o percentual de linf?citos-TNF+ nas culturas estimuladas com PMA. A an?lise cromatogr?fica em fase gasosa acoplada a espectrometria de massas (CG/EM) revelou como principais constituintes no ?leo essencial as subst?ncias ?-pineno (7,51%), limoneno (5,9%), ?xido de cariofileno (13,59%), 1,2 ep?xido humuleno (8,41%) e 1,6-humulanodien-3-ol (17,71%). O extrato dicloromet?nico de A. fastigiatum, na concentra??o 20 ?g/mL, n?o apresentou toxicidade aos leuc?citos do sangue perif?rico humano e reduziu o percentual de linf?citos-TNF-?+ e linf?citos-IFN-?+ nas culturas estimuladas com o PMA. Este extrato foi fracionado em coluna de Sephadex LH-20 (150 g). Na an?lise de citocinas, a fra??o 10 (AFDM 10), na concentra??o 10 ?g/mL, demonstrou efeito anti-inflamat?rio in vitro reduzindo a frequ?ncia de linf?citos-TNF-?+ e a prolifera??o de linf?citos estimulados com PHA (fitohemaglutinina). Sugere-se que parte da atividade anti-inflamat?ria de A. fastigiatum se d? pela inibi??o que os constituintes da planta promovem sobre a ativa??o de leuc?citos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2013. / ABSTRACT
Ageratum fastigiatum is a plant used in folk medicine as topical anti-inflammatory and analgesic agent. However, few studies have been conducted with this plant in order to detail the molecular mechanisms involved in its biological activities. Therefore, main purpose of the present study was to investigate the anti-inflammatory activity of the essential oil and leaf rinse dichloromethane of A. fastigiatum extract. The cytotoxicity of the compounds was assessed by means of trypan blue exclusion. The essential oil at 5x10-3 e 1x10-2 ?L/ did not present cytotoxic activity. These two concentrations were used for investigating the anti-inflammatory potential of the essential oil, by analyzing the pro-inflammatory (TNF-? and IFN-?) and anti-inflammatory (IL-10) cytokine profiles on human leukocyte cultures stimulated and non-stimulated with PMA (phorbol myristate acetate). The data obtained by flow cytometry demonstrated that both concentrations decreased the percentage of lymphocytes ?TNF-? + on the cell cultures stimulated with PMA. In the non-stimulated cell cultures there were no changes in the percentage of IL-10+ cells. The gas phase chromatography coupled with mass spectrometry analysis (GC/MS) revealed that the main compounds present in oil samples were: ?-pinene (7.51%), limonene (5.9%), caryophyllene oxide (13.59%), 1,2 humulene epoxide (8.41%) and 1,6-humulanodien-3-ol (17.71%). The dichloromethane extract of A. fastigiatum, obtained from the washing leaves, at the concentration 20 ?g/mL, did not presented cytotoxicic action on the human peripheral blood leukocytes. Following the same protocol performed to cytokines analysis used before, we observed that the dichloromethane A. fastigiatum extract also decreased the percentage of lymphocytes-TNF-? + and IFN-?+ on the cell cultures stimulated with PMA. This extract after fractionated on Sephadex LH-20 . The dichloromethane A. fastigiatum fraction (AFDM 10) at 10 ?g/mL reducing the frequency of lymphocytes-TNF-?+. Furthermore AFDM 10 decreasing the proliferation of lymphocytes stimulated with PHA (phytohemagglutinin). It is concluded that part of the anti-inflammatory activity of A. fastigiatum is give by inhibiting the plant constituents on promoting the activation of leukocytes.
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Composi??o qu?mica e efeito sobre mediadores inflamat?rios de prepara??es de partes a?reas de Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob (arnica-do-campo) in vitroAlmeida, Val?ria Gomes de 30 January 2015 (has links)
?rea de concentra??o: Ci?ncias fisiol?gicas. / Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-04T17:58:44Z
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Previous issue date: 2015 / A Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob., (Asteraceae) ? nativa da flora brasileira, e sua prepara??o alco?lica tem sido utilizada de forma t?pica como anti-inflamat?rio na medicina popular. Tendo em vista a aus?ncia de estudos acerca da bioatividade e composi??o qu?mica dessa esp?cie, o objetivo do presente estudo foi avaliar o efeito de prepara??es de partes a?reas de P.brasiliensis sobre par?metros inflamat?rios, in vitro, e realizar a an?lise dos seus constituintes qu?micos. A triagem fitoqu?mica dos caules, flores e folhas da planta foi realizada por meio de rea??es cromog?nicas, fluorog?nicas e de precipita??o e sugeriu a presen?a de saponinas, terpenos, taninos e flavonoides nas partes a?reas da planta. Os extratos etan?licos de caules (PBETca), flores (PBETfl) e folhas (PBETfo) foram avaliados por cromatografia l?quida de alta efici?ncia acoplada a detector de arranjo de diodos (CLAE-DAD). Essa an?lise sugeriu a presen?a de flavonoides nos extratos, especialmente das classes das flavonas e flavon?is. Os ?leos essenciais de flores (PBOEfl) e de folhas (PBOEfo) foram analisados por Cromatografia Gasosa acoplada a Espectr?metro de Massas (CG-EM), sendo poss?vel a identifica??o de 22 compostos no PBOEfl e 27 no PBOEfo, dentre os quais, o ?-pineno, limoneno, ?-pineno, sabineno, ?xido de cariofileno e E-cariofileno est?o presentes em maior valor percentual. Foram confeccionadas culturas de 4h de sangue total para avaliar o efeito hemol?tico dos extratos, determinado pela densidade ?ptica do sobrenadante em 540 nm. Houve indu??o de hem?lise em culturas tratadas com PBETca ou PBETfo em concentra??es superiores a 100 ?g/mL. O extrato PBETfl induziu hem?lise em culturas na concentra??o final de 200 ?g/mL. A toxicidade dos produtos naturais ?s C?lulas Mononucleares do Sangue Perif?rico (PBMC) humano ap?s 24 h e 5 dias, foi avaliada por citometria de fluxo, utilizando Azul de Tripan ou Iodeto de Prop?deo. O efeito dos produtos naturais sobre a produ??o de TNF-?, IFN-? e IL-2 foi determinado por ELISA do sobrenadante de culturas de 24 h, ap?s est?mulo com Miristato Acetato de Forbol (PMA) + Ionomicina. Todos os extratos e ?leos essenciais reduziram a produ??o de TNF-?. A s?ntese de IFN-? foi inibida pelo extrato PBETfo e pelo ?leo essencial PBOEfl. Todos os extratos etan?licos e o PBOEfl, inibiram a produ??o de IL-2. O PBETfo foi fracionado por meio de parti??o em solventes com polaridade crescente e todas as fra??es foram eficientes em inibir a s?ntese de TNF-?, por PBMC, ap?s 24 h de cultura. Somente a fra??o hex?nica reduziu a s?ntese de IFN-? e IL-2. Foram confeccionadas culturas de sangue total estimuladas com lipopolissacar?deo (LPS) para investiga??o do efeito dos ?leos essenciais sobre a express?o de
COX-2, por citometria de fluxo. Os ?leos essenciais n?o interferiram na express?o de COX-2. Nossos achados sugerem que o efeito anti-inflamat?rio atribu?do ? P.brasiliensis na medicina popular pode, pelo menos em parte, ser devido ? inibi??o da produ??o de citocinas pr?-inflamat?rias por linf?citos e mon?citos humanos. ? poss?vel que flavonoides das subclasses das flavonas e flavon?is possam contribuir para a atividade biol?gica dos extratos etan?licos de P. brasiliensis. Muitos monoterpenos e sesquiterpenos presentes nos ?leos essenciais de flores e folhas de P. brasiliensis j? tiveram a atividade anti-inflamat?ria descrita em outros trabalhos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / ABSTRACT
The Pseudobrickellia brasiliensis (Spreng) R. M. King & H. Rob., (Asteraceae) is native from Brazilian flora, and its alcoholic preparation has been used topically as anti-inflammatory in folk medicine. Considering the lack of studies about the bioactivity and chemical composition of this specie, the aim of this study was to evaluate the effect of P. brasiliensis aerial parts preparations on in vitro inflammatory parameters, and perform analysis of its chemical constituents. The plant stems, flowers and leaves phytochemical screening was carried out by chromogenic, fluorogenic and precipitation reactions and suggested the presence of saponins, terpenes, tannins and flavonoids in the aerial parts of the plant. The ethanolic extracts of stems (PBETca), flowers (PBETfl) and leaves (PBETfo) were evaluated by high performance liquid chromatography coupled to diode array detector (HPLC-DAD). These analyses suggested the presence of flavonoids in the extracts, especially the flavones and flavonols subclasses. The flowers (PBOEfl) and leaves (PBOEfo) essential oils were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), and it was possible to identify 22 and 27 compounds in PBOEfl and PBOEfo, respectively. Among the compounds the ?-pinene, limonene, ?-pinene, sabinene, caryophyllene oxide and E-caryophyllene were present in high porcentage. A 4h culture of whole blood was prepared to evaluate the hemolytic effect of the extracts, determined by supernatant optical density at 540 nm. There were hemolysis induction in cultures treated with PBETca or PBETfo at concentrations exceeding 100 ?g/mL. The extract PBETfl induced hemolysis in cultures at a final concentration of 200 ?g/mL. The natural products toxicity to human Peripheral Blood Mononuclear Cells (PBMCs) after 24 h and 5 days were assessed by flow cytometry using trypan blue or propidium iodide. The effect of non-toxic concentrations of the natural products on TNF-?, IFN-? and IL-2 production was determined by ELISA of 24h supernatants culture after stimulation with phorbol myristate acetate (PMA) + Ionomycin. All extracts and essential oils reduced the TNF-? production. The IFN-? synthesis was inhibited by PETfo extract and PBOEfl essential oil.The PBETfo and PBOEfl inhibited the IFN-? synthesis. The PBOEfl and all ethanolic extracts inhibited IL-2 production. The PBETfo was fractionated by solvent partitioning of increasing polarity and all fractions were effective in inhibiting the TNF-? synthesis by PBMC after 24 h culture. Only the hexane fraction reduced the IFN-? and IL-2 synthesis. Whole blood cultures stimulated with lipopolysaccharide (LPS) were prepared to investigate the effect of essential
oils on COX-2 expression by flow cytometry. Essential oils did not affect the expression of COX-2. Our findings suggest that the anti-inflammatory effect attributed to P.brasiliensis in folk medicine can, at least in part, be due to the inhibition of proinflammatory cytokines production by human lymphocytes and monocytes. It is possible that flavonoids of flavones and flavonols subclasses may contribute to the biological activity of ethanolic extracts of P. brasiliensis. Many monoterpenes and sesquiterpenes present in essential oils of P. brasiliensis flowers and leaves already had the anti-inflammatory activity described in other studies.
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Efeito do extrato etan?lico de Ageratum fastigiatum (Gardn.) R. M. King et H. Rob. e de sua fra??o sobre a produ??o de citocinas e prolifera??o de linf?citos humanos, in vitroCosta, Lucas de Abreu 13 May 2016 (has links)
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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.
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Um novo inibidor de tripsina multifuncional de sementes de Erythrina velutina: caracterizacao e propriedades farmacologicasMachado, Richele Jana?na de Ara?jo 24 October 2012 (has links)
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Previous issue date: 2012-10-24 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / A sepse ? uma doen?a complexa, de alta gravidade e durante o seu
desenvolvimento pode ocorrer eventos inflamat?rios graves, que podem levar o
paciente ao ?bito. Inibidores de peptidases isolados de sementes de leguminosas
est?o sendo estudados devido suas variadas atividades biom?dicas ou
farmacol?gicas. Nesse sentido, o presente estudo consistiu na avalia??o das
atividades multifuncionais do inibidor de tripsina presente nas sementes de Erythrina
velutina (EvTIb). Dois inibidores de tripsina foram purificados por fracionamento com
sulfato de am?nio (30-60%), cromatografia de afinidade em tripsina-sepharose e
cromatografia de fase reversa. Os inibidores purificados foram denominados de
EvTIa e EvTIb e suas massas moleculares determinadas por espectrometria de
massa por electrospray (ESI) foram: 19.228,16 Da e 19.210,48 Da, respectivamente.
Dentre eles, o EvTIb apresentou maior atividade espec?fica para tripsina e maior
percentual de rendimento de extra??o, sendo, portanto, o escolhido para as an?lises
adicionais. Os fragmentos desse inibidor gerados por tratamento enzim?tico com
tripsina e pepsina foram analisados por espectrometria de massa MALDI-ToF-ToF,
permitindo elucidar sua estrutura prim?ria parcial. O EvTIb exibiu uma atividade de
inibi??o do tipo n?o-competitiva para a tripsina com IC50 de 22 nmol.L-1 e constante
de inibi??o (Ki) de 10 nmol.L-1. Al?m da atividade de inibi??o para tripsina, EvTIb
tamb?m inibiu o fator Xa e a elastase neutrof?lica, por?m n?o inibiu trombina,
quimotripsina ou peptidase 3. EvTIb se manteve est?vel a varia??o de pH e
temperatura. EvTIb n?o foi citot?xico e n?o inibiu o crescimento bacteriano de
Escherichia coli e Staphylococcus aureus. EvTIb foi capaz de prolongar o tempo de
coagula??o de maneira dose-dependente. Finalmente, observou-se que EvTIb foi
capaz de restabelecer os n?veis hemost?ticos, inibir a migra??o leucocit?ria em
camundongos s?pticos e diminuir a secre??o de TNF-α e n?o teve a??o sobre a
libera??o de IL-6. Por outro lado, EvTIb promoveu um aumento na libera??o de IFN-
α e IL-12. Esses dados sugerem que EvTIb pode constituir em uma nova mol?cula
com potencial de uso no tratamento da sepse e doen?as relacionadas ? coagula??o
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Avalia??o da atividade anti-inflamat?ria do extrato aquoso, fra??es e compostos identificados nos frutos de Hancornia Speciosa Gomes (Apocynaceae)R?go, Manoela Torres do 19 June 2015 (has links)
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Previous issue date: 2015-06-19 / Hancornia speciosa Gomes (Apocynaceae), conhecida popularmente
por ?mangabeira?, ? usada na medicina popular para o tratamento de desordens
inflamat?rias, hipertens?o, dermatites, diabetes, doen?as hep?ticas e
desordens estomacais. No que diz respeito aos frutos de H. speciosa, a
etnobot?nica indica o uso principalmente para o tratamento de inflama??o e
tuberculose, entretanto n?o h? relatos de estudos que comprovem sua poss?vel
atividade biol?gica. O estudo tem como objetivo avaliar a atividade antiinflamat?ria
do extrato aquoso dos frutos de H. speciosa Gomes. O extrato
aquoso foi preparado por decoc??o, posteriormente submetido a um
fracionamento por meio de parti??o l?quido-l?quido e os metabolitos
secund?rios identificados por Cromatografia l?quida de alta efici?ncia acoplada
ao detector de arranjos de diodos (CLAE-DAD) e Cromatografia l?quida
acoplada ao detector de arranjo de diodo e ao espectr?metro de massa (CLDAD-EM).
As propriedades anti-inflamat?rias do extrato aquoso, fra??es
diclorometano (CH2Cl2), acetato de etila (AcOEt) e n-butanol (n-BuOH), bem
como a rutina e o ?cido clorog?nico foram investigadas usando modelos in vivo
e in vitro. Testes in vivo compreenderam edema de orelha induzido por xilol,
onde foi mensurado a forma??o do edema; peritonite induzida por carragenina
onde foi avaliado os leuc?citos totais em 4h e bolsa de ar induzida por zimosam
que foi mensurado a contagem total de leuc?citos e contagem diferencial em 6,
24 e 48 horas. Os testes in vitro avaliaram os n?veis de citocinas IL-1?, IL-6, IL-
12 e TNF-? usando ELISA a partir do modelo de peritonite induzido por
carragenina. Os resultados demonstraram a presen?a de rutina e ?cido
clorog?nico no extrato aquoso dos frutos de H. speciosa por CLAE-DAD e CLDAD-EM.
O extrato aquoso e fra??es, bem como a rutina e ?cido clorog?nico
inibiram significativamente o edema de orelha induzido por xilol e reduziram a
migra??o celular em modelos animais, tanto na peritonite induzida por
carragenina como na bolsa de ar induzida por zimosam. Al?m disso, foi
observada a redu??o dos n?veis de express?o de citocinas IL-1?, IL-6, IL-12 e
TNF-?. Este ? o primeiro estudo que demonstra o efeito anti-inflamat?rio do
extrato aquoso dos frutos de Hancornia speciosa contra diferentes agentes
inflamat?rios em modelos animais, sugerindo que suas mol?culas bioativas,
especialmente rutina e ?cido clorog?nico contribuem, em parte, para o efeito
anti-inflamat?rio do extrato aquoso. Estes resultados suportam a utiliza??o de
H. speciosa na medicina popular e revelam que o extrato aquoso apresenta um
potencial terap?utico para o desenvolvimento de uma droga herbal com
propriedades anti-inflamat?rias. / Hancornia speciosa Gomes (Apocynaceae), popularly known as
?mangabeira?, has been used in folk medicine to treat inflammatory disorders,
hypertension, dermatitis, diabetes, liver diseases and stomach disorders.
Regarding the Hancornia speciosa fruits, the ethnobotany indicates its use
especially for treating inflammation and tuberculosis. However, no study has
been done so far to prove such biological activities. The objective was
evaluation anti-inflammatory activity from the fruits of Hancornia speciosa
Gomes (mangabeira). Aqueous extract was prepared by decoction,
subsequently submitted the liquid-liquid fractionation. The secondary
metabolites were identified by high performance liquid chromatography coupled
with detector diode array (HPLC-DAD) and liquid chromatography diode array
detector coupled with mass spectrometry (LC-DAD-MS). The anti-inflammatory
properties of the aqueous extract, dichloromethane (CH2Cl2), ethyl acetate
(EtOAc) and n-butanol (n-BuOH) fractions of the fruits from H. speciosa, as well
as rutin and chlorogenic acid were investigated using in vitro and in vivo
models. In vivo tests comprised the xylene-induced ear edema that was
measured the formation of edema, carrageenan-induced peritonitis was
evaluated the total leukocytes at 4h and zymosan-induced air pouch was
measured the total leukocytes and differential cell count at 6, 24 and 48 hours,
whereas in vitro tests were evaluated levels of cytokines IL-1?, IL-6, IL-12 and
TNF-? using ELISA obtained of carrageenan-induced peritonitis model. The
results showed the presence of rutin and chlorogenic acid were detected in the
aqueous extract from H. speciosa fruits by HPLC-DAD and LC-DAD-ME.
Furthermore, the aqueous extracts and fractions, as well as rutin and
chlorogenic acid significantly inhibited the xilol-induced ear edema and reduced
cell migration in the animal models such as carrageenan-induced peritonitis and
zymosan-induced air pouch. In addition, reduced levels of cytokines IL-1?, IL-6,
IL-12 and TNF-? were observed. This is the first study that demonstrated the
anti-inflammatory effect of aqueous extract from Hancornia speciosa fruits
against different inflammatory agents in animal models, suggesting that their
bioactive molecules, especially rutin and chlorogenic acid contributing, at least
in part, to the anti-inflammatory effect of aqueous extract. These findings
support the widespread use of Hancornia speciosa in popular medicine and
demonstrate that this aqueous extract has therapeutic potential for the
development of a herbal drugs with anti-inflammatory properties.
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Spondias tuberosa Arr. (UMBU): estudo fitoqu?mico e avalia??o do potencial anti-inflamat?rioSiqueira, Emerson Michell da Silva 26 February 2015 (has links)
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Previous issue date: 2015-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Spondias tuberosa Arruda (Anacardiaceae) ? uma ?rvore frut?fera conhecida
popularmente no Brasil como umbuzeiro, tapereba ou umbu. A planta ? nativa e
end?mica do Brasil, distribu?da por todos estados do nordeste brasileiro. Essa esp?cie
tem grande import?ncia na medicina popular do semi-?rido nordestino, onde ? utilizada
principalmente para tratar condi??es inflamat?rias, problemas digestivos assim como
infec??es virais e bacterianas. No entanto, apesar da vasta utiliza??o na medicina
tradicional, existem poucos estudos farmacol?gicos e fitoqu?micos que d?em
embasamento cient?fico a sua utiliza??o popular. Diante disto, este trabalho buscou
caracterizar os marcadores qu?micos e avaliar a atividade anti-inflamat?ria do extrato
das folhas de Spondias tuberosa. O extrato das folhas foi preparado por macera??o em
etanol:?gua (70:30, v/v). A avalia??o do perfil fitoqu?mico do extrato foi realizada
preliminarmente por Cromatografia em Camada Delgada e em seguida por
Cromatografia L?quida de Alta Efici?ncia. Na an?lise por Cromatografia em Camada
Delgada foi observada a presen?a dos flavonoides rutina e isoquercitrina, o que foi
corroborado pela an?lise por Cromatografia L?quida de Alta Efici?ncia que ainda
indicou a presen?a dos ?cidos fen?licos, ?cido clorog?nico e ?cido caf?ico.
Adicionalmente, foi desenvolvido e validado um m?todo por Cromatografia L?quida de
Alta Efici?ncia segundo as normas da RDC 899/2003 da ANVISA e ICH Guidelines
2005, para avaliar o teor dos compostos identificados no extrato. Para avalia??o do
efeito anti-inflamat?rio foram utilizados os modelos de peritonite e de edema de pata
induzidos por carragenina, administra??o via intraperitoneal, em camundongos. A
avalia??o do efeito anti-inflamat?rio in vivo demonstrou que o extrato das folhas, nas
concentra??es de 125, 250 e 500 mg/kg, apresentou uma diminui??o do influxo de
leuc?citos para o local da inflama??o, do di?metro do edema e do n?vel da enzima
mieloperoxidase, quando comparado ao f?rmaco utilizado como controle dexametasona
(2 mg/kg, via i.p.). Em suma, em nosso trabalho foi verificado que o extrato
hidroetan?lico das folhas de Spondias tuberosa apresentou atividade anti-inflamat?ria
importante nos dois modelos ensaiados, o que torna a planta uma potencial esp?cie para
este uso, justificando o seu uso popular. No que se refere ? caracteriza??o de
marcadores, os compostos fen?licos, ?cido clorog?nico, ?cido caf?ico, rutina e
isoquercitrina, identificados no extrato hidroetan?lico das folhas de Spondias tuberosa
podem ser utilizados no controle de qualidade da mat?ria-prima e de extratos obtidos
com a esp?cie. / Spondias tuberosa Arruda (Anacardiaceae) is a fruitful tree popularly known as
umbuzeiro, tapereba or umbu. It is a native and endemic species from Brazil,
widespread in Brazilian Northeast. The species is important in folk medicine of the
semi-arid Northeast, where it is mainly used to treat various inflammatory conditions,
digestive problems as well as viral and bacterial infections. However, despite the
common use in folk medicine, there are scarce pharmacological and phytochemicals
studies that afford scientific evidence to its popular use. Therefore, this study aimed to
characterize the chemical markers in S. tuberosa leaves extract, obtained by maceration
ethanol:water (70:30, [v/v]), and evaluate its anti-inflammatory potential in vivo. The
phytochemical profile in TLC analysis suggested the occurence of the flavonoids rutin
and isoquercitrin. HPLC analysis enabled us to confirm the presence of flavonoids and
also, were detected the phenolic acids, chlorogenic acid and caffeic acid. In addition
was developed and validated a HPLC method to evaluate the content of the identified
compounds in S. tuberosa leaves extract according to RDC 899/2003 of ANVISA and
ICH Guidelines 2005. In order to evaluate the anti-inflammatory potential of S.
tuberosa leaves extract, the peritonitis and paw edema models induced by carrageenan
were used, administration i.p. in mice. The results highlighted the anti-inflammatory
property in vivo at 125, 250 and 500 mg/kg since a decrease in leukocyte influx to the
site of inflammation, diameter of the edema and the level of myeloperoxidase were
observed when compared to the drug control dexamethasone (2 mg/kg, i.p. route).
Taken together, the results pointed out S. tuberosa as a potential species for developing
phytotherapic derivatives in according to its popular use. With regard to the
characterization markers, chlorogenic acid, caffeic acid, rutin and isoquercitrin were
identified and quantified in Spondias tuberosa leaves extract so they could be used in
quality control analyses of the raw material and extracts of this species.
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Mecanismos envolvidos com as atividades antinociceptiva, antiedematog?nica e antiinflamat?ria do flavonoide majorit?rio das infloresc?ncias de Kalanchoe pinnata (Lam.) Pers. (folha da fortuna) / Mechanisms underlying the antinociceptive, antiedematogenic, and anti-inflammatory activity of the main flavonoid from Kalanchoe pinnata flowers (Lam.) Pers. (fortune sheet)Ferreira, Raquel Teixeira 30 March 2016 (has links)
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Previous issue date: 2016-03-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Kalanchoe pinnata (KP) is popularly used for treating inflammatory diseases. This study
investigated the antinociceptive and anti-inflammatory potential of the KP flower aqueous
extract (KPFE), the ethyl acetate (EtOAcF), butanol (BuOHF) and aqueous (AqF) fractions
and the main flavonoid [quercetin 3-O-?-L-arabinopyranosyl (1??) ?-L-rhamnopyranoside]
(KPFV) of KP, as well as possible mechanisms of action. Swiss albino mice, male (25-35g)
were pretreated with KPFE (30-300 mg/kg, s.c.) producing dose-related inhibition of acetic
acid-induced writhing (ID50 164.8 mg/kg), and the subcutaneous administration of KPFE (300
mg/kg), EtOAcF (12 mg/kg), BuOHF (15 mg/kg) or AqF (210 mg/kg) reduced leukocyte
migration on carrageenan-induced pleurisy in mice at 56.1%, 47.3%, 39.6% and 43.1%,
respectively. At doses of up to 240 times smaller than KPFE, KPFV (0.3-3.0 mg/kg, s.c.), also
inhibited leukocyte migration (ID50 1.78 mg/kg). In croton oil-induced ear edema in mice,
KPFE (3-30 mg/kg, s.c.) and KPFV (0.3-3.0 mg/kg, s.c.) presented dose-related antiedematogenic
activity (KPFE - ID50 4.3 mg/kg, KPFV - ID50 0.76 mg/kg). Treatment with
KPFE reduced the MPO and TNF-? concentration in the pleural exudates on carrageenaninduced
pleurisy test, while KPFV inhibited both isoforms of cyclooxygenase (COX), with
IC50 3.8 x 10-5 M (22.1 ?g/mL) for COX-1 and a maximum COX-2 inhibition of 43.5% (IC50
>8.4 x10-5M or 50 ?g/mL). The selectivity index (COX-1 IC50/COX-2 IC50) was <0.44. These
results indicate that KPFE and KPFV produced anti-inflammatory and antinociceptive activity
through the COX inhibition and TNF-? reduction. This study reveals that the main flavonoid
in KP flowers and leaves plays an important role in the ethnomedicinal use of this species for
healing inflammatory processes / Kalanchoe pinnata (KP) ? popularmente utilizada para tratamento de doen?as inflamat?rias.
Este estudo investigou o potencial antinociceptivo e anti-inflamat?rio do extrato aquoso das
infloresc?ncias de KP (EAFL), a fra??o acetato de etila (FAcOEt), a fra??o butan?lica
(FBuOH), a fra??o aquosa (FAq) e o flavonoide majorit?rio [quercetina 3-O-?-Larabinopiranosil
(1??) ?-L-ramnopiranos?deo] (KPFV) de KP, assim como poss?veis
mecanismos de a??o. Camundongos albinos, machos (25-35g) foram pr?-tratados com EAFL
(30-300 mg/kg, s.c.), produzindo inibi??o dose dependente das contor??es induzidas por ?cido
ac?tico (DI50 164,8 mg/kg), e a administra??o subcut?nea de EAFL (300 mg/kg), FAcOEt (12
mg/kg) ou FBuOH (15 mg/kg) reduziu a migra??o leucocit?ria na pleurisia induzida por
carragenina em camundongos em 56,1%, 47,3%, 39,6% e 43,1%, respectivamente. Em doses
at? 240 vezes menores que o EAFL, KPFV (0,3-3,0 mg/kg, s.c.), tamb?m foi capaz de inibir a
migra??o leucocit?ria (DI50 1,78 mg/kg). No teste do edema de orelha induzido por ?leo de
cr?ton em camundongos, EAFL (3-30 mg/kg, s.c.) e KPFV (0,3-3,0 mg/kg, s.c.) apresentaram
atividade antiedematog?nica dose dependente (EAFL - DI50 4,3 mg/kg, KPFV - DI50 0,76
mg/kg). O tratamento com EAFL reduziu a concentra??o de MPO e de TNF-? no exsudato
pleural obtido do teste da pleurisia induzida por carragenina, enquanto o KPFV inibiu ambas
as isoformas de cicloxigenase (COX), com CI50 3,8 x 10-5M (22,1 ?g/mL) para COX-1 e um
m?ximo de inibi??o da COX-2 de 43,5% (CI50 >8,4 x10-5M ou 50 ?g/mL). O ?ndice de
seletividade (COX-1 CI50/COX-2 CI50) foi < 0,44. Estes resultados indicam que EAFL e
KPFV produziram atividade anti-inflamat?ria e antinociceptiva envolvendo a inibi??o da
COX e a redu??o de TNF-?. Este estudo revela que o principal flavonoide presente nas
infloresc?ncias e folhas de KP possui papel importante no uso etnomedicinal desta esp?cie
nos processos de natureza inflamat?ria
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Caracteriza??o do efeito antinociceptivo e anti-inflamat?rio do polissacar?deo extra?do da levedura Kluyveromyces marxianusOliveira, Renata Freitas de Araujo 26 May 2015 (has links)
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Previous issue date: 2015-05-26 / The yeasts produce different types of metabolites and macromolecules with huge biotechnological potential. Among them, the Kluyveromyces marxianus, species attracts industrial and economic interests for presenting certain qualities, such as thermal tolerance, high growth rates, broad substrate spectrum and modulation of immune response. This work investigated the effects of treatment with the polysaccharide extracted from the Kluyveromyces marxianus yeast (poly322) in pain and inflammation experimental models. The poly322 pharmacological properties were evaluated with writhing, formalin, nociception (manifested by CFA), tail flick, hot-plate and paw edema tests. The cytokine levels were determined by ELISA and the effects on motor performance were evaluated by rotarod and open field tests. The pretreatment with poly322 resulted in reduced nociception induced by acetic acid, CFA and formalin (late phase). Furthermore, animals treated with poly322 exhibited a significant reduction in mechanical hypernociception, paw edema, and local IL-6 levels increase induced by carrageenan. In contrast, treatment with poly322 did not alter the thermal stimulus response threshold in the tail flick and hot plate tests, indicating no central action. Confirming the specificity of the action suggested by nociceptive tests, the treatment with poly322 did not induce motor impairment. These results demonstrate that the poly322 has a potent antinociceptive and anti-inflammatory effects, possibly mediated by the inhibition of the production and/or release of the IL-6 cytokine. / As leveduras produzem diferentes tipos de metab?litos e macromol?culas com enorme potencial biotecnol?gico. Dentre essas, destaca-se a Kluyveromyces marxianus, esp?cie que desperta interesse industrial e econ?mico por apresentar qualidades, como a toler?ncia t?rmica, altas taxas de crescimento, amplo espectro de substrato e modula??o da resposta imune. O presente estudo investigou os efeitos do tratamento com o polissacar?deo extra?do da levedura Kluyveromyces marxianus (poly322) em modelos experimentais de dor e inflama??o. As propriedades farmacol?gicas do poly322 foram avaliadas nos testes de contor??es abdominais, formalina, nocicep??o manifesta por CFA, retirada da cauda, placa quente e edema de pata. Os n?veis de citocinas foram determinados por ELISA e os efeitos sobre o desempenho motor foram avaliados pelos testes do cil?ndro girat?rio e campo aberto. O pr?-tratamento com o poly322 resultou na redu??o da hipernocicep??o induzida pelo ?cido ac?tico, CFA e formalina (segunda fase). Al?m disso, os animais tratados com o poly322 exibiram uma redu??o significativa no edema de pata e no aumento local dos n?veis de IL-6 induzidos pela carregenina. Em contraste, o tratamento com o poly322 n?o alterou o limiar de resposta a est?mulo t?rmico nos testes de retirada da cauda e placa quente, indicando aus?ncia de a??o central. Confirmando a especificidade da a??o sugerida pelos testes nociceptivos, o tratamento com o poly322 n?o induziu comprometimento motor. Os resultados demonstram que o poly322 possui um potente efeito antinociceptivo e anti-inflamat?rio, possivelmente mediado pela inibi??o da produ??o e/ou libera??o da citocina IL-6.
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