Avaliação do efeito da ouabaína na viabilidade e modulação de citocinas em timócitos e linfócitos expostos a radiação ultravioleta

• Autor(a) principal: Alves, Anne Kaliery de Abreu
• Data de Publicação: 2018
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Biblioteca Digital de Teses e Dissertações da UFPB
• Texto Completo: https://repositorio.ufpb.br/jspui/handle/123456789/13130
• topic: Inflamação; Timócitos; Linfócitos; Radiação; Inflammation; Thymocytes; Lymphocytes; Radiation; CNPQ::CIENCIAS BIOLOGICAS

Ouabain is a cardiac glycoside discovered in human plasma in 1991 (HAMLYN et al., 1991). This endogenous substance can inhibit Na + / K + -ATPase and has been extensively studied for its ability to interfere with various mechanisms that regulate and maintain homeostasis. Thus, the aim of this study was to evaluate in vitro and in vivo ouabain regarding cell viability and the profile of pro-inflammatory cytokines in thymus and mesenteric lymph node cells exposed to UV radiation. Therefore, Swiss albinus mice were used conducting cell culture from the thymus and the mesenteric lymph node. These cells were treated with ouabain 100, 10, 1 µM and 100 nM in a plaque of 96 wells, and also with a pre-treatment with ouabain 0.56 mg/kg during three days via intraperitoneal injection. The cells obtained from the maceration of the organs were centrifuged, resuspended and counted. The concentration was adjusted to 4x105céls/ml with PBS and plated for exposure to UVC and UVB for 2 minutes. After the stimuli, the RPMI medium supplemented with 10% of SFB is added and cells are kept in culture for 6 to 24 hours. Viability was analysed with the MTT 5 mg/mL and with the pro-inflammatory cytokines TNF-a and IL-6, as determined by the method ELISA. Our results showed that after 6h and 24h the culture in the presence of ouabain in different concentrations 100,10, 1 µM and100 nM, only in the 100 nM the cells were protected from cell death, this result was observed for the 24h culture of thymocytes and lymphocytes that were stimulated by UVC, but this was not observed for UVB stimulation. In these cells, the pre-treatment with 0.56 mg/Kg ouabain in the 6 hours culture presented a cytoprotective effect on the thymocytes and mesenteric lymph node lymphocytes exposed to UVC and UVB. Besides these effects on viability, ouabain in the concentration of 100 nM increased TNF-a and reduced IL-6. Nevertheless, the pre-treatment with 0.56 mg/Kg did not modulate these cytokines compared to the group receiving radiation. These results contribute to the understanding of the ouabain action over important parameters of the inflammatory process, viability and cytokines, triggered by ultraviolet radiation.