Atividade anti-Inflamatória da Marinobufagina em modelos in vivo e in vitro

• Autor(a) principal: Carvalho, Deyse Cristina Madruga
• 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/123456789/13137
• topic: Marinobufagina; Esteroides cardiotônicos; Inflamação; Marinobufagenin; Cardiotonic steroids; Inflammation; CNPQ::CIENCIAS BIOLOGICAS::FISIOLOGIA

Cardiotonic steroids are natural compounds capable of inhibiting the Na+/K+-ATPase. These steroids, such as marinobufagenin and ouabain, have been identified as endogenous substances present in mammalian plasma. Our group has been demonstrating the role of ouabain in the immune response by describing and characterizing its anti-inflammatory effect. However, there are still no reports in the literature about the marinobufagenin in this regard. Therefore, the aim of this work was to analyze, in vivo and in vitro, the role of marinobufagenin in inflammation. Initially, Swiss mice were treated intraperitoneally (i.p.) with marinobufagenin at the dose 0,56 mg/kg, the same used in ouabain experiments (LEITE et al., 2015), for three consecutive days. One hour after the last day of treatment, animals were stimulated with zymosan (2 mg/mL) intraperitoneally, in order to induce peritoneum inflammation. After 4 h, peritoneal fluid was collected and used for counting cells by optical microscopy and for quantification of proinflammatory cytokines (IL-1ß, IL-6 and TNFa) by immunoenzymatic assay (ELISA). Zymosan, as expected, induced increased cell migration and proinflammatory cytokine levels in the peritoneum. On the other hand, marinobufagenin treatment was able to reduce the total number of cells into the peritoneal cavity by reducing migration of polymorphonuclear cells. In addition, this steroid was able to reduce IL-1ß and IL-6 levels. This effect appears to be independent of TNF-a, since its levels were not affected. In addition, in vitro effect marinobufagenin was evaluated by culture of peritoneal macrophages with zymosan (2 mg/mL) or lipopolysaccharide (LPS) (1 µg/mL) stimulation. In macrophage culture with zymosan stimulation, it was observed that different concentrations of marinobufagenin (10, 100, 1,000 and 10,000 nM) did not interfere in viability of peritoneal macrophages and only the lowest concentration was able to reduce proinflammatory cytokines IL -1ß, IL-6 and TNF-a levels. In macrophage culture with the LPS stimulation, it was observed that all marinobufagenin concentrations were able to reduce nitric oxide (NO) production in these cells. Thus, from the analysis of the results, it can be suggested that there is evidence that marinobufagenin has an anti-inflammatory role in vivo and in vitro. Therefore, this work is pioneer in characterizing immunological role of the cardiotonic steroid marinobufagenin.