Análise do proteoma salivar e degradação da histatina 1 e 5 na saliva de sujeitos com paralisia cerebral e gengivite

• Ano de defesa: 2020
• Autor(a) principal: Gutierrez, Gabriela Mancia de
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Cruzeiro do Sul; Brasil; Programa de Pós Graduação em Odontologia; Cruzeiro do Sul
• Palavras-chave em Português: Saliva; Espectrometria de Massas; Gengivite; Paralisia cerebral; Proteômica; CNPQ::CIENCIAS DA SAUDE::ODONTOLOGIA
• Link de acesso: https://repositorio.cruzeirodosul.edu.br/handle/123456789/919

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.