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Previous issue date: 2010-05-07 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Seeds from legumes including the Glycine max are known to be a rich source of protease inhibitors. The soybean Kunitz trypsin inhibitor (SKTI) has been well characterised and has been found to exhibit many biological activities. However its effects on inflammatory diseases have not been studied to date. In this study, SKTI was purified from a commercial soy fraction, enriched with this inhibitor, using anion exchange chromatography Resource Q column. The purified protein was able to inhibit human neutrophil elastase (HNE) and bovine trypsin. . Purified SKTI inhibited HNE with an IC50 value of 8 ?g (0.3 nM). At this concentration SKTI showed neither cytotoxic nor haemolytic effects on human blood cell populations. SKTI showed no deleterious effects on organs, blood cells or the hepatic enzymes alanine amine transferase (ALT) and aspartate amino transferase (AST) in mice model of acute systemic toxicity. Human neutrophils incubated with SKTI released less HNE than control neutrophils when stimulated with PAF or fMLP (83.1% and 70% respectively). These results showed that SKTI affected both pathways of elastase release by PAF and fMLP stimuli, suggesting that SKTI is an antagonist of PAF/fMLP receptors. In an in vivo mouse model of acute lung injury, induced by LPS from E. coli, SKTI significantly suppressed the inflammatory effects caused by elastase in a dose dependent manner. Histological sections stained by hematoxylin/eosin confirmed this reduction in inflammation process. These results showed that SKTI could be used as a potential pharmacological agent for the therapy of many inflammatory diseases / Sementes de leguminosas s?o conhecidas como uma rica fonte de inibidores de proteinases, destacando-se dentre estes o inibidor de tripsina da soja (SKTI) que ? uma prote?na amplamente estudada e caracterizada para muitas propriedades biol?gicas. Entretanto seus efeitos aplicados a desordens inflamat?rias ainda s?o pouco conhecidos. SKTI foi purificado ? partir de uma fra??o comercial de soja atrav?s de cromatografia de troca ani?nica em Resource Q. A prote?na purificada foi capaz de inibir a elastase de neutr?filos humanos (ENH) e a tripsina bovina. O valor da sua IC50 foi de 8 μg.mL-1 (0.3 nM) e nessa concentra??o o SKTI n?o foi capaz de provocar efeitos hemol?ticos ou citot?xicos sobre as popula??es celulares sangu?neas humanas. Por meio do modelo de toxicidade sist?mica aguda, utilizando camundongos, tamb?m n?o foram observados efeitos delet?rios sobre ?rg?os, c?lulas sangu?neas e altera??es nos n?veis das enzimas hep?ticas aspartato amino transferase (AST) e alanina amino transferase (ALT). Neutr?filos humanos incubados com SKTI na concentra??o de 0.3 nM apresentaram uma diminui??o da libera??o de ENH quando estimulados pelos ativadores PAF e fMLP (83,1% e 70 %, respectivamente). Estes resultados mostram que o SKTI foi capaz de afetar ambas as vias PAF/fMLP de libera??o de ENH, sugerindo esta prote?na como um poss?vel antagonista dos receptores PAF/fMLP. Modelos in vivo de inj?ria pulmonar aguda mediante estimula??o por LPS de Escherichia. coli demonstraram uma supress?o significativa dos eventos inflamat?rios atribu?dos ? atividade elast?sica de forma dose dependente. Cortes histol?gicos corados por hematoxilina e eosina confirmaram a diminui??o da inflama??o tecidual. Estes resultados sugerem que o SKTI pode ser indicado como um potencial agente farmacol?gico na terapia de muitas doen?as inflamat?rias
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.ufrn.br:123456789/12557 |
| Date | 07 May 2010 |
| Creators | Ribeiro, Jannison Karlly Cavalcante |
| Contributors | CPF:29706106391, http://lattes.cnpq.br/7890362793618911, Rocha, Hugo Alexandre de Oliveira, CPF:76111830449, http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4799567J8&dataRevisao=null, Vasconcelos, Ilka Maria, CPF:31109128304, http://lattes.cnpq.br/0103197383336584, Sales, Maur?cio Pereira de |
| Publisher | Universidade Federal do Rio Grande do Norte, Programa de P?s-Gradua??o em Bioqu?mica, UFRN, BR, Bioqu?mica; Biologia Molecular |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Repositório Institucional da UFRN, instname:Universidade Federal do Rio Grande do Norte, instacron:UFRN |
| Rights | info:eu-repo/semantics/openAccess |
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