Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório do Centro Universitário Braz Cubas |
Texto Completo: | https://repositorio.cruzeirodosul.edu.br/handle/123456789/919 |
Resumo: | This study aimed to analyze the differences in the protein profile and to evaluate the histol proteolysis profile (his) 1 and 5 of the whole saliva of adolescents with cerebral palsy (CP) and gingivitis compared to healthy ones with and without gingivitis, using mass spectrometry. 24 individuals were selected and divided into 3 groups: G1 - CP with gingivitis, G2 - without CP and without gingivitis, G3 - without CP and with gingivitis. Gingival index (GI) and simplified oral hygiene index (SOHI) were collected, in addition to total unstimulated saliva. One pool was performed for each group and the peptides from the digestion of salivary proteins were subjected to liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) and the proteins were identified using the database data from human Uniprot. Protein degradation was carried out by adding his 1 and his 5 synthetic to the diluted supernatant of whole saliva (1:10) and incubation at 37ºC for times 0, 0.5, 1.5, 4, 6, 8, 24 and 48 hours. Then, electrophoresis analyzes were performed on cationic polycrylamide gel (PAGE) and measurement of the band density (BD) (%) of the images obtained. The analysis of data referring to BD during different periods of time of the means of the individuals of each group and between the gingival health parameters were analyzed with One-way ANOVA or Kruskal-Wallis test. Multilevel Poisson Regression analysis was performed to assess the factors that influence the BD percentage of histatine degradation (considering the Subject level) (α = 5%). The groups showed no significant difference in salivary flow, but there was difference in gingival indexes for groups that presented gingivitis (p <0.001). 487 proteins were identified; 183 proteins from pool 1, 288 from pool 2 and 189 from pool 3. Of the exclusive proteins, 115 were from pool 1, 214 from pool 2 and 72 from pool 3. In common between pools 1 and 3 were 14 proteins. Through Poisson regression analysis, the Subject having CP and gingivitis influences the degradation of his 1 and his 5 (p <0.001), the age of the Subject influences the degradation of his 5, the older the greater the degradation (p <0.001) and that the IG influences the degradation of his 1 (p <0.001). It can be concluded that his 1 is more resistant to proteolysis than his 5, the degradation of his 1 and his 5 occurs more quickly in individuals with CP and gingivitis. Haptoglobin and toll-like receptor 9 were found exclusively in pool 1 and have three immunological functions: immune response, antimicrobial activity and inflammatory response. This approach provides a new insight into the proteomic profile of saliva during gingival inflammation, and some proteins found exclusively in the saliva of individuals with CP and gingivitis (pool 1) were previously identified as potential biomarkers for periodontitis, indicating that individuals with CP may present greater risk for the progression of periodontitis. |
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Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengiviteSalivaEspectrometria de MassasGengiviteParalisia cerebralProteômicaCNPQ::CIENCIAS DA SAUDE::ODONTOLOGIAThis study aimed to analyze the differences in the protein profile and to evaluate the histol proteolysis profile (his) 1 and 5 of the whole saliva of adolescents with cerebral palsy (CP) and gingivitis compared to healthy ones with and without gingivitis, using mass spectrometry. 24 individuals were selected and divided into 3 groups: G1 - CP with gingivitis, G2 - without CP and without gingivitis, G3 - without CP and with gingivitis. Gingival index (GI) and simplified oral hygiene index (SOHI) were collected, in addition to total unstimulated saliva. One pool was performed for each group and the peptides from the digestion of salivary proteins were subjected to liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) and the proteins were identified using the database data from human Uniprot. Protein degradation was carried out by adding his 1 and his 5 synthetic to the diluted supernatant of whole saliva (1:10) and incubation at 37ºC for times 0, 0.5, 1.5, 4, 6, 8, 24 and 48 hours. Then, electrophoresis analyzes were performed on cationic polycrylamide gel (PAGE) and measurement of the band density (BD) (%) of the images obtained. The analysis of data referring to BD during different periods of time of the means of the individuals of each group and between the gingival health parameters were analyzed with One-way ANOVA or Kruskal-Wallis test. Multilevel Poisson Regression analysis was performed to assess the factors that influence the BD percentage of histatine degradation (considering the Subject level) (α = 5%). The groups showed no significant difference in salivary flow, but there was difference in gingival indexes for groups that presented gingivitis (p <0.001). 487 proteins were identified; 183 proteins from pool 1, 288 from pool 2 and 189 from pool 3. Of the exclusive proteins, 115 were from pool 1, 214 from pool 2 and 72 from pool 3. In common between pools 1 and 3 were 14 proteins. Through Poisson regression analysis, the Subject having CP and gingivitis influences the degradation of his 1 and his 5 (p <0.001), the age of the Subject influences the degradation of his 5, the older the greater the degradation (p <0.001) and that the IG influences the degradation of his 1 (p <0.001). It can be concluded that his 1 is more resistant to proteolysis than his 5, the degradation of his 1 and his 5 occurs more quickly in individuals with CP and gingivitis. Haptoglobin and toll-like receptor 9 were found exclusively in pool 1 and have three immunological functions: immune response, antimicrobial activity and inflammatory response. This approach provides a new insight into the proteomic profile of saliva during gingival inflammation, and some proteins found exclusively in the saliva of individuals with CP and gingivitis (pool 1) were previously identified as potential biomarkers for periodontitis, indicating that individuals with CP may present greater risk for the progression of periodontitis.Esse estudo teve como objetivo analisar as diferenças no perfil de proteínas e avaliar o perfil da proteólise da histatina (his) 1 e 5 da saliva total de adolescentes com paralisia cerebral (PC) e gengivite comparados a saudáveis com e sem gengivite, utilizando espectrometria de massas. Foram selecionados 24 indivíduos, os quais foram divididos em 3 grupos: G1 - PC com gengivite, G2 - sem PC e sem gengivite, G3 - sem PC e com gengivite. Foram coletados o índice gengival (IG) e índice de higiene oral simplificado (IHO-S), além da saliva total não estimulada. Foram realizados um pool para cada grupo e os peptídeos provenientes da digestão das proteínas salivares foram submetidos a cromatografia líquida seguida de ionização por electrospray e espectrometria de massas em tandem (LC-ESI-MS/MS) e as proteínas foram identificadas usando o banco de dados de humanos Uniprot. A degradação proteica foi realizada pela adição de his 1 e his 5 sintéticas ao sobrenadante da saliva total diluído (1:10) e incubação a 37ºC pelos tempos 0, 0,5, 1,5, 4, 6, 8, 24 e 48 horas. Em seguida, foram realizadas as análises de eletroforese em gel de poliacrilamida catiônica (PAGE) e mensuração da densidade das bandas (DB) (%) das imagens obtidas. A análise dos dados referentes à DB durante diferentes períodos de tempos das médias dos índivíduos de cada grupo e entre os parâmetros da saúde gengival foi realizada com One-way ANOVA ou teste de Kruskal-Wallis. Para avaliação dos fatores que influenciam a porcentagem de DB da degradação das histatinas foi realizada análise de Regressão de Poisson Multinível (considerando o nível do Sujeito) (α=5%). Os grupos não apresentaram diferença significativa no fluxo salivar, mas houve diferença nos índices gengivais para os grupos que possuíam gengivite (P < 0,001). Foram identificadas 487 proteínas; 183 proteínas do pool 1, 288 do pool 2 e 189 do pool 3. Das proteínas exclusivas, 115 foram do pool 1, 214 do pool 2 e 72 do pool 3. Em comum entre os pools 1 e 3 foram 14 proteínas. Por meio da análise de regressão de Poisson o Sujeito ter PC e gengivite influencia na degradação da his 1 e his 5 (P < 0,001), a idade do Sujeito influencia na degradação da his 5, quanto mais velho maior degradação (P < 0,001) e que o IG influencia na degradação da his 1 (P < 0,001). Pode-se concluir que a his 1 é mais resistente à proteólise do que a his 5, a degradação das his 1 e his 5 ocorre mais rapidamente em indivíduos com PC e gengivite. A haptoglobin e toll-like receptor 9 foram encontradas exclusivamentes no pool 1 e possuem as 3 três funções imunológicas: resposta imune, atividade antimicrobiana e resposta inflamatória. Essa abordagem fornece uma nova visão sobre o perfil proteômico da saliva durante a inflamação gengival e, algumas proteínas encontradas exclusivamente na saliva de indivíduos com PC e gengivite (pool 1) foram previamente apontadas como potenciais biomarcadores para periodontites, indicando que indivíduos com PC podem apresentar maior risco para a progressão da periodontite.Universidade Cruzeiro do SulBrasilPrograma de Pós Graduação em OdontologiaCruzeiro do SulSantos, Maria Teresa Botti Rodrigues07562092850http://lattes.cnpq.br/0781261124888655Santos, Maria Teresa Botti Rodrigues07562092850http://lattes.cnpq.br/0781261124888655Ortega, Adriana de Oliveira Lirahttp://lattes.cnpq.br/8035653789643276Douek, Débora HellerAmado, Pamela Pontes Penashttp://lattes.cnpq.br/1979789761590545Alves, Levy Anderson Cesarhttp://lattes.cnpq.br/4583482989383583Gutierrez, Gabriela Mancia de2020-08-24T22:05:03Z2020-04-242020-08-24T22:05:03Z2020-03-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfGUTIERREZ, G. M. Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite. 100 f. Tese (Doutorado em Odontologia). São Paulo: Universidade Cruzeiro do Sul, 2020.https://repositorio.cruzeirodosul.edu.br/handle/123456789/919porinfo:eu-repo/semantics/openAccessreponame:Repositório do Centro Universitário Braz Cubasinstname:Centro Universitário Braz Cubas (CUB)instacron:CUB2020-08-24T22:07:05Zoai:repositorio.cruzeirodosul.edu.br:123456789/919Repositório InstitucionalPUBhttps://repositorio.brazcubas.edu.br/oai/requestbibli@brazcubas.edu.bropendoar:2020-08-24T22:07:05Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB)false |
dc.title.none.fl_str_mv |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
title |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
spellingShingle |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite Gutierrez, Gabriela Mancia de Saliva Espectrometria de Massas Gengivite Paralisia cerebral Proteômica CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
title_short |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
title_full |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
title_fullStr |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
title_full_unstemmed |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
title_sort |
Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite |
author |
Gutierrez, Gabriela Mancia de |
author_facet |
Gutierrez, Gabriela Mancia de |
author_role |
author |
dc.contributor.none.fl_str_mv |
Santos, Maria Teresa Botti Rodrigues 07562092850 http://lattes.cnpq.br/0781261124888655 Santos, Maria Teresa Botti Rodrigues 07562092850 http://lattes.cnpq.br/0781261124888655 Ortega, Adriana de Oliveira Lira http://lattes.cnpq.br/8035653789643276 Douek, Débora Heller Amado, Pamela Pontes Penas http://lattes.cnpq.br/1979789761590545 Alves, Levy Anderson Cesar http://lattes.cnpq.br/4583482989383583 |
dc.contributor.author.fl_str_mv |
Gutierrez, Gabriela Mancia de |
dc.subject.por.fl_str_mv |
Saliva Espectrometria de Massas Gengivite Paralisia cerebral Proteômica CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
topic |
Saliva Espectrometria de Massas Gengivite Paralisia cerebral Proteômica CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA |
description |
This study aimed to analyze the differences in the protein profile and to evaluate the histol proteolysis profile (his) 1 and 5 of the whole saliva of adolescents with cerebral palsy (CP) and gingivitis compared to healthy ones with and without gingivitis, using mass spectrometry. 24 individuals were selected and divided into 3 groups: G1 - CP with gingivitis, G2 - without CP and without gingivitis, G3 - without CP and with gingivitis. Gingival index (GI) and simplified oral hygiene index (SOHI) were collected, in addition to total unstimulated saliva. One pool was performed for each group and the peptides from the digestion of salivary proteins were subjected to liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) and the proteins were identified using the database data from human Uniprot. Protein degradation was carried out by adding his 1 and his 5 synthetic to the diluted supernatant of whole saliva (1:10) and incubation at 37ºC for times 0, 0.5, 1.5, 4, 6, 8, 24 and 48 hours. Then, electrophoresis analyzes were performed on cationic polycrylamide gel (PAGE) and measurement of the band density (BD) (%) of the images obtained. The analysis of data referring to BD during different periods of time of the means of the individuals of each group and between the gingival health parameters were analyzed with One-way ANOVA or Kruskal-Wallis test. Multilevel Poisson Regression analysis was performed to assess the factors that influence the BD percentage of histatine degradation (considering the Subject level) (α = 5%). The groups showed no significant difference in salivary flow, but there was difference in gingival indexes for groups that presented gingivitis (p <0.001). 487 proteins were identified; 183 proteins from pool 1, 288 from pool 2 and 189 from pool 3. Of the exclusive proteins, 115 were from pool 1, 214 from pool 2 and 72 from pool 3. In common between pools 1 and 3 were 14 proteins. Through Poisson regression analysis, the Subject having CP and gingivitis influences the degradation of his 1 and his 5 (p <0.001), the age of the Subject influences the degradation of his 5, the older the greater the degradation (p <0.001) and that the IG influences the degradation of his 1 (p <0.001). It can be concluded that his 1 is more resistant to proteolysis than his 5, the degradation of his 1 and his 5 occurs more quickly in individuals with CP and gingivitis. Haptoglobin and toll-like receptor 9 were found exclusively in pool 1 and have three immunological functions: immune response, antimicrobial activity and inflammatory response. This approach provides a new insight into the proteomic profile of saliva during gingival inflammation, and some proteins found exclusively in the saliva of individuals with CP and gingivitis (pool 1) were previously identified as potential biomarkers for periodontitis, indicating that individuals with CP may present greater risk for the progression of periodontitis. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-08-24T22:05:03Z 2020-04-24 2020-08-24T22:05:03Z 2020-03-03 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
GUTIERREZ, G. M. Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite. 100 f. Tese (Doutorado em Odontologia). São Paulo: Universidade Cruzeiro do Sul, 2020. https://repositorio.cruzeirodosul.edu.br/handle/123456789/919 |
identifier_str_mv |
GUTIERREZ, G. M. Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite. 100 f. Tese (Doutorado em Odontologia). São Paulo: Universidade Cruzeiro do Sul, 2020. |
url |
https://repositorio.cruzeirodosul.edu.br/handle/123456789/919 |
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por |
language |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Cruzeiro do Sul Brasil Programa de Pós Graduação em Odontologia Cruzeiro do Sul |
publisher.none.fl_str_mv |
Universidade Cruzeiro do Sul Brasil Programa de Pós Graduação em Odontologia Cruzeiro do Sul |
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reponame:Repositório do Centro Universitário Braz Cubas instname:Centro Universitário Braz Cubas (CUB) instacron:CUB |
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Centro Universitário Braz Cubas (CUB) |
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Repositório do Centro Universitário Braz Cubas - Centro Universitário Braz Cubas (CUB) |
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bibli@brazcubas.edu.br |
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