Análise funcional do microRNA-769-3P no processo fibrosante hepático e sua correlação com elementos pró-inflamatórios em células estreladas humanas

• Autor(a) principal: Ramos, Letícia Ferreira [UNESP]
• Data de Publicação: 2017
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/156459; http://www.athena.biblioteca.unesp.br/exlibris/bd/capelo/2018-04-26/000898124.pdf
• topic: Biologia molecular; MicroRNAs; Células hepáticas; Figado; Figado - Doenças; Homeostase

The liver plays several roles in cellular homeostasis, but, unfortunately, high number of people are accomited by hepatic problems, characterizing them as a serious public health problem. One ordinary characteristic of such pathologies is fibrosis, which is stablished due to the intense healing response to the damaged tissue resulting in the accumulation of extracellular matrix components. If the lesion is continuous, pro-inflammatory mechanisms are established compromising the organ functions. In the liver, the cells responsible for the synthesis of extracellular matrix elements are the hepatic stellate cells. In health liver, such cells store vitamin A in lipid droplets. In pro-fibrotic livers, the cell loses its star-shaped conformation and the lipid droplets. In general, the inflammatory response is characteristic of liver injury, and pro-inflammatory stimuli are the result of arachidonic acid metabolism in the prostaglandin synthesis pathway, mediated by the action of cyclooxygenase proteins. Recent studies point to a connection between cyclooxygenase 2 protein and lipid droplet forming enzymes (acyl-CoA synthetases). In parallel, effective therapies against fibrosis are still elusive and the characterization of new molecules that support the development of alternative therapies is aimed. In this context, microRNAs comes as a promising molecule. In this study the microRNA-769-3p was functionally investigated focusing in its metabolic correlation with the lipid metabolism and pro-inflammatory processes. For the development of this work, LX-2 lineage liver cells were cultured for the analysis of lipid droplets by Oil Red O, for analysis of gene expression by RT-PCR and qRT-PCR, and immunofluorescence and ELISA analyzes for the identification of some important proteins in the fibrogenic process. Cellular transfection with either miRNA-769-3p mimic or inhibitor change their metabolism and conduce to the reappearance of lipid droplets. A more...