Validação de um painel de biomarcadores proteicos no plasma seminal de adolescentes com e sem varicocele

• Autor(a) principal: Belardin, Larissa Berloffa [UNIFESP]
• Data de Publicação: 2016
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=3613154; https://repositorio.unifesp.br/handle/11600/46162
• topic: Adolescent; Seminal plasma proteins; Proteomics; Testis; Varicocele; Adolescente; Proteínas de plasma seminal; Proteômica; Testículo; Varicocele

Objective: To verify the seminal levels of potential biomarkers for testicular changes in adolescent varicocele. The specific objectives were to verify if the presence of varicocele and seminal alterations can alter the levels of the following proteins: Cab45 (45 kDa calcium-binding protein); LEFTY1 (Left-right determination factor 1); DNase I (deoxyribonuclease-1); PAP2-Alpha (Lipid phosphate phosphohydrolase 1); IFGBP7 (Insulin-like growth factor binding protein-7); IGHG3 (Ig gamma-3 chain C region); CRISP-3 (Cysteine-rich secretory protein 3). Method: This observational study included 61 adolescents, aged 15 to 17 years old with full sexual maturity (Tanner V), which were divided into 3 groups: control (without varicocele, n=20); VSN (with varicocele, normal semen analysis, n=22); and VSA (with varicocele and altered semen analysis, n=19). The adolescents collected one seminal sample after 2 to 5 days of ejaculatory abstinence. After semen liquefaction semen was centrifuged, in order to separate the seminal plasma from cellular constituents. Total protein in each sample was then quantified, and a volume correspondent to 50 ?g of protein for each patient was used for Western blotting for the follow proteins: Cab 45, LEFTY1, DNase I, PAP2-Alpha, IGFBP7, IGHG3 and CRISP-3 (all were normalized to the protein Epididymis Secretory Sperm Binding protein 67p Li [DJ-1]). Results: We observed lower sperm concentration (million/mL) and morphology (% normal sperm) in the VSA group when compared to the other groups. There was a decrease in the levels of Cab45 in both groups of adolescents with varicocele (VSN and VSA), with a lowest value in the VSA group. DNase I levels were lower in the group of adolescents with varicocele and altered semen analysis (VSA). There was an overexpression of IGFBP7 in the groups with varicocele when compared with control groups, and an overexpression of the two isoforms of CRISP-3 protein in the VSA group. Conclusion: Proteomics of seminal plasma reflects changes in testicular function in the adolescent varicocele. The levels of proteins LEFTY1, PAP2-Alpha, IGHG3 are not altered in the presence of a varicocele. Cab45 levels are decreased in the presence of varicocele; DNase I levels are decreased in the presence of seminal alterations and varicocele. Conversely, IGFBP7 is a positive marker of varicocele, and CRISP-3 is a positive marker of varicocele and altered semen quality. These results may help in the future to determine the medical management for early surgical intervention in varicocele.