AVALIAÇÃO IN VITRO DE LEITE DE CAPRINO EXPRESSANDO LISOZIMA HUMANA RECOMBINANTE

• Ano de defesa: 2015
• Autor(a) principal: MENEZES, JOSÉ NILSON RODRIGUES DE
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual do Ceará
• Palavras-chave em Português: Biotecnologia
• Link de acesso: https://siduece.uece.br/siduece/trabalhoAcademicoPublico.jsf?id=88277

<!--[if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:TrackMoves/> <w:TrackFormatting/> <w:DoNotShowRevisions/> <w:DoNotPrintRevisions/> <w:DoNotShowMarkup/> <w:DoNotShowComments/> <w:DoNotShowInsertionsAndDeletions/> <w:DoNotShowPropertyChanges/> <w:HyphenationZone>21</w:HyphenationZone> <w:PunctuationKerning/> <w:ValidateAgainstSchemas/> <w:SaveIfXMLInvalid>false</w:SaveIfXMLInvalid> <w:IgnoreMixedContent>false</w:IgnoreMixedContent> <w:AlwaysShowPlaceholderText>false</w:AlwaysShowPlaceholderText> <w:DoNotPromoteQF/> <w:LidThemeOther>PT-BR</w:LidThemeOther> <w:LidThemeAsian>X-NONE</w:LidThemeAsian> <w:LidThemeComplexScript>X-NONE</w:LidThemeComplexScript> <w:Compatibility> <w:BreakWrappedTables/> <w:SnapToGridInCell/> <w:WrapTextWithPunct/> <w:UseAsianBreakRules/> <w:DontGrowAutofit/> <w:SplitPgBreakAndParaMark/> <w:DontVertAlignCellWithSp/> <w:DontBreakConstrainedForcedTables/> <w:DontVertAlignInTxbx/> <w:Word11KerningPairs/> <w:CachedColBalance/> </w:Compatibility> <w:DoNotOptimizeForBrowser/> <m:mathPr> <m:mathFont m:val="Cambria Math"/> <m:brkBin m:val="before"/> <m:brkBinSub m:val="&#45;-"/> <m:smallFrac m:val="off"/> <m:dispDef/> <m:lMargin m:val="0"/> <m:rMargin m:val="0"/> <m:defJc m:val="centerGroup"/> <m:wrapIndent m:val="1440"/> <m:intLim m:val="subSup"/> <m:naryLim m:val="undOvr"/> </m:mathPr></w:WordDocument> </xml><![endif]--> <div class="WordSection1"> <p class="MsoBodyText" style="margin-top:11.6pt;margin-right:53.3pt;margin-bottom: 0cm;margin-left:85.1pt;margin-bottom:.0001pt;text-align:justify;line-height: 150%">O leite humano contém em média 400 &#956;g/mL de lisozima que é considerada e uma defesa natural contra bactérias, vírus, parasitas e fungos. O crescimento diferencial de bactérias no leite de animais transgênicos indica que a lisozima expressa no leite é capaz de agir de forma antimicrobiana reduzindo a velocidade do crescimento de bactérias <i style="mso-bidi-font-style:normal">in vivo</i>. Sendo assim este estudo teve como objetivo geral Avaliar <i style="mso-bidi-font-style:normal">in vitro </i>o leite de caprino expressando lisozima humana recombinante e como objetivos específicos: 1) Caracterizar o leite de cabra transgênica quanto à expressão de lisozima humana recombinante; 2) Avaliar a migração e a viabilidade de células epiteliais intestinais linhagem IEC-18, na presença de leite de caprino transgênico contendo lisozima humana. Neste estudo foram coletadas amostras de leite da cabra transgênica para lisozima humana fundadora da linhagem <i style="mso-bidi-font-style:normal">Lisa</i>, da Universidade de Fortaleza, bem como o leite caprino não transgênico. Os leites foram comparados ao leite materno humano e a uma solução aquosa de lisozima humana recombinante comercial. Foi realizada a análise físico-química, e os padrões analisados encontram-se dentro dos requisitos mínimos preconizados pelo Ministério da Agricultura, Pecuária e Abastecimento <span style="letter-spacing:-.2pt">(MAPA) </span>para qualidade do leite de cabra destinado ao consumo humano. Com os resultados da identificação da lisozima por <i style="mso-bidi-font-style:normal">Western Blot</i>, podemos confirmar a presença da proteína no leite transgênico e como esperado no leite materno humano. A atividade antibacteriana foi avaliada através do teste de difusão, onde a sensibilidade da cepa de <i style="mso-bidi-font-style:normal">Micrococcus luteus </i>foi testada em relação à presença da hLZ, onde a formação do halo de inibição envolta do poço em que foi colocada a amostra, e nos indica o potencial antimicrobiano do leite transgênico. Assim a cepa de <i style="mso-bidi-font-style:normal">M. luteus </i>testada, apresentou alta sensibilidade à presença da lisozima, ou seja, o leite transgênico apresentou um poder bactericida quando comparado aos grupos controle e ao leite não transgênico. O leite transgênico apresentou características semelhantes ao leite não transgênico de cabras controle, comprovando que a presença da lisozima não altera ou prejudica a integridade da mucosa intestinal, pelo contrário, foram obtidos resultados em que o leite transgênico ajudou na migração <span style="letter-spacing:-.15pt">celular, </span>como também manteve a viabilidade celular em sua presença. A migração celular na presença de <i style="mso-bidi-font-style:normal">Enterococcus faecalis </i>foi<span style="letter-spacing:.55pt"> </span>superior<span style="letter-spacing:.5pt"> </span>quando<span style="letter-spacing:.55pt"> </span>o<span style="letter-spacing:.5pt"> </span>leite<span style="letter-spacing:.55pt"> </span>transgênico<span style="letter-spacing:.6pt"> </span>foi<span style="letter-spacing:.45pt"> </span>adicionado<span style="letter-spacing:.5pt"> </span>ao<span style="letter-spacing:.5pt"> </span>meio.<span style="letter-spacing:.7pt"> </span>Portanto,<span style="letter-spacing:.6pt"> </span>concluímos<span style="font-size:11.0pt;line-height:150%;font-family:&quot;Arial&quot;,&quot;sans-serif&quot;; mso-fareast-font-family:Arial;mso-ansi-language:PT-BR;mso-fareast-language: PT-BR;mso-bidi-language:PT-BR"> <br/></span></p></div><p class="MsoBodyText" style="margin-top:5.05pt;margin-right:53.3pt;margin-bottom: 0cm;margin-left:85.1pt;margin-bottom:.0001pt;text-align:justify;line-height: 150%">que o leite transgênico contém a proteína hLZ com quantidade próxima a do leite materno humano, apresentando-se físico e quimicamente dentro dos padrões estabelecidos para o consumo humano, possui atividade antibacteriana, não alterou o nível de expressão da proteína Zo-1 e da IL-6, se tornando um potencial nutracêutico para o combate a diarreia e a desnutrição, contribuindo para as pessoas com diarreia e desnutrição para os profissionais de saúde que trabalham nesta área, pesquisadores e para a sociedade como um<span style="letter-spacing:-.55pt"> </span>todo.</p> <p class="MsoBodyText"><span style="font-size:18.0pt;mso-bidi-font-size:12.0pt">&nbsp;</span></p> <p class="MsoBodyText" style="margin-top:.05pt;margin-right:53.3pt;margin-bottom: 0cm;margin-left:85.1pt;margin-bottom:.0001pt;text-align:justify;line-height: 150%"><b style="mso-bidi-font-weight:normal">Palavras-chave: </b>Leite. Lisozima humana. <span style="mso-ansi-language:EN-US" lang="EN-US">Cabras transgênicas. Diarreia. 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