Caracterização do perfil de linfócitos T no desenvolvimento do diabetes mellitus tipo 2 em ratos Goto-kakizaki

• Ano de defesa: 2020
• Autor(a) principal: Manoel, Richelieau
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Cruzeiro do Sul; Brasil; Departamento 1; Programa de Pós-Graduação Interdisciplinar em Ciências da Saúde; Cruzeiro do Sul
• Palavras-chave em Português: Resistência à insulina; Células T reguladoras; células Th1; GLUT1; CNPQ::CIENCIAS DA SAUDE
• Link de acesso: https://repositorio.cruzeirodosul.edu.br/handle/123456789/906

Peripheral resistance to insulin (RI) and type 2 diabetes mellitus (DM2) have a high prevalence in obese patients. However, a large number of non-obese patients have DM2. Goto-Kakizaki (GK) rats develop a well-defined RI and DM2 without presenting obesity. The characterization of the immune response of these animals and in humans in these conditions is not yet elucidated in the literature. Therefore, the aim of this study is to characterize the process of T lymphocyte differentiation during the development of DM2 in Goto-Kakizaki rats, in order to determine whether changes in subpopulations of Th cells may be associated with the genesis of DM2 in absence of obesity. Changes in the expression pattern of genes involved with cell differentiation of lymphocytes from GK rats at 21, 60 and 120 days after birth were evaluated. Lymphocytes were isolated from the mesenteric lymph nodes of these animals after euthanasia. In lymphocytes it was evaluated: expression of GLUT1 in T lymphocyte membrane by flow cytometry; proliferative capacity of T lymphocyte and analysis of transcription factor and cytokine gene expression (Foxp3, GATA3 and T-bet, TGF-beta, TNF-alpha, IL-10 and INFgamma) involved in the process of differentiation of T lymphocytes by real-time PCR. We observed in the glucose tolerance test and insulin tolerance test, an increase in blood glucose levels in the GK group, as well as in the area under the curve, showing possible insulin resistance in animals with 21, 60 and 120 days. Regarding the expression of GLUT1, there is an increase in animals from GK group at all ages in nonstimulated condition. When we evaluated the ratio of GLUT1 + cells in the condition of stimulation with PMA and Ionomycin by baseline, we observed a higher value in the GK group compared to the Wistar group (Control). In the analysis of gene expression involved with differentiation of lymphocytes, we observed lower values of GATA-3 in the GK group at all ages compared to Wistar (Control). For Foxp3 (Treg cells), we observed a lower expression in the rats with 21 days from GK group compared to the Wistar. For the T-bet (Th1 cells), we observed an increase for the GK group, compared to Wistar with only 120 days. For TNF-α, we observed an increase in GK at 60 and 120 days and for INF-γ, we observed greater expression for GK at all ages. In conclusion, we can state that GK animals with 21 days have a reduction in Th2 antiinflammatory response markers (Gata-3 and IL-10) and Treg (Foxp3 and IL-10), indicating that reduction of immunosuppressor mechanisms early in the animal's life may favor pro-inflammatory responses in later stages of life. This is because, the expression of Th1 profile cytokines is increased only in animals with 60 and 120 days.