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Previous issue date: 2017-11-24 / Animal venoms are one of the richest sources of biologically active substances found in nature and such prerogative has been confirmed in pharmacological and biochemical studies of proteins (enzymes), peptides, bioactive amines, and other compounds isolated from snake venoms. In this context, the purpose of the present study was to evaluate the antibacterial and antitumor potential of the individual venoms "yellow" (Cdr68 and Cdr69) and "white" varieties of the Amazonian rattlesnake Crotalus durissus ruruima. The evaluation of the antimicrobial activity of the crude venoms was performed by the disc diffusion technique against gram-positive (Staphylococcus aureus and S. epidermidis) and gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa). The yellow venoms presented antibacterial activity against gram-positive bacteria Staphylococcus aureus. The cytotoxic action of the venoms was evaluated using the following cell lines SK-Mel 103 (melanoma), MCF-7 (breast adecarcinoma), HCT-116 (colorectal carcinoma) and MCR-5 (human fibroblast). Cdr68 and Cdr69 venoms were cytotoxic to all tumor lines but were more potent for the colorectal carcinoma (HCT-116) with IC50 of 1.8 μg / mL and 1.3 μg / mL for Cdr68 and Cdr69 respectively. The white variety venoms were not cytotoxic to the tested strains. The chromatographic profiles of Cdr68, Cdr69, Cdr110 and Cdr173 by molecular exclusion showed four major peaks. The isolated fractions were submitted to tests of coagulant, phospholipase A2, cytotoxic and antibacterial activity, all activities were present in Peak II of Cdr68, and in Peaks I (cytotoxic) and II of Cdr69. Peaks II of both venoms were submitted to Reverse Phase Chromatography. The FRP2 peaks of the Reverse Phase of the venoms presented phospholipase activity, cytotoxic against the strain HCT-116 in concentration of 100 μg / mL and antibacterial against S. aureus. The mass of this peak was approximately 14 kDa, compatible with PLA2. It is interesting to note that the total venom presented higher cytotoxic potential than the isolated fractions, showing a possible synergistic effect among the venom constituents. / Os venenos animais constituem uma das mais ricas fontes de substâncias biologicamente ativas encontradas na natureza e tal prerrogativa tem sido confirmada em estudos farmacológicos e bioquímicos, realizados com proteínas (enzimas), peptídeos, aminas bioativas, dentre outros compostos, isolados de venenos de serpentes. Neste contexto, o objetivo do presente trabalho foi avaliar o potencial antibacteriano e antitumoral dos venenos individuais, variedade “amarela” (Cdr68 e Cdr69) e “branca” (Cdr110 e Cdr173) da cascavel Amazônica Crotalus durissus ruruima. A avaliação da atividade antimicrobiana dos venenos foi realizada pela técnica de difusão do disco contra as bactérias gram-positivas (Staphylococcus aureus e S. epidermidis) e gram-negativas (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa). Os venenos amarelos apresentaram atividade antibacteriana contra a bactéria gram-positiva Staphylococcus aureus. A ação citotóxica dos venenos foi avaliada utilizando as seguintes linhagens celulares SK-Mel 103 (melanoma), MCF-7 (adecarcinoma de mama), HCT-116 (carcinoma colorretal) e MCR-5 (fibroblasto humano). Os venenos Cdr68 e Cdr69 foram citotóxicos para todas as linhagens tumorais, mas foram mais potentes para a linhagem de carcinoma colorretal (HCT-116) com CI50 de 1,8 μg/mL e 1,3 μg/mL para Cdr68 e Cdr69, respectivamente. Os venenos variedade branca não foram citotóxicos para as linhagens testadas. Os perfis cromatográficos de Cdr68, Cdr69, Cdr110 e Cdr173 de Exclusão Molecular apresentaram quatro picos principais. As frações isoladas foram submetidas aos testes de atividade coagulante, fosfolipásica A2, citotóxica e antibacteriana, todas as atividades estavam presentes no Pico II de Cdr68, e nos Picos I (citóxica) e II do Cdr69. Os picos II de ambos os venenos foram submetidos à Cromatografia de Fase Reversa. Os picos FRP2 da Fase Reversa dos venenos apresentaram atividade fosfolipásica, citotóxica frente à linhagem HCT-116 na concentração de 100 μg/mL e antibacteriana contra S. aureus. A massa deste pico foi de aproximadamente 14 kDa, compatível com PLA2. Interessante notar, que o veneno total apresentou maior potencial citotóxico do que as frações isoladas, mostrando um possível efeito sinérgico entre os constituintes do veneno.
| Identifer | oai:union.ndltd.org:IBICT/oai:http://localhost:tede/6471 |
| Date | 24 November 2017 |
| Creators | Santos, Ilia Gilmara Carvalho dos, 92-99200-3719 |
| Contributors | ppgbiotec@ufam.edu.br, Santos, Maria Cristina dos, Dias, Consuelo Latorre Fortes |
| Publisher | Universidade Federal do Amazonas, Programa de Pós-Graduação em Biotecnologia, UFAM, Brasil, Instituto de Ciências Biológicas |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFAM, instname:Universidade Federal do Amazonas, instacron:UFAM |
| Rights | http://creativecommons.org/licenses/by-nc-nd/4.0/, info:eu-repo/semantics/openAccess |
| Relation | 1984485651082134923, 500 |
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