Mechanistic and functional analyses of the microRNA-1914-5p in the nonalcoholic hepatic pro-steatotic processes in cell culture model

Abstract

Nowadays, nonalcoholic fat liver diseases (NAFLD) are considered a serious health problem worldwide. Such diseases are described by an excessive fat storage in the liver and despite several different ethiologic agents, the molecular aspects that supports NAFLD progression to its worse clinical aspects have to be described. In previously FAPESP grant, our research group described the miR-1914-5p as an effective element that modulates the lipid metabolism in hepatic stellate cells (HSCs) LX-2 under the heptapetide angiotensin-(1-7) treatment. These cells are direct connected with the hepatic fibrogenesis. Thus, the description of the mechanisms by wich the miR-1914-5p acts in the steatotic processes is relevant, and will probably contribute to new therapeutic strategies. To perform the analyses, steatotic cellular models will be set up, using coculture of hepatocytes and LX-2 as models, which will be transfected with the miR-1914-5p mimic or inhibitor molecules. Next, cellular and molecular assays will be performed to evaluate the effect of such small molecules in cellular physiopathology, based on the analyses of global triacylglicerol and cholesterol lipids profile in the cultures by gas chromatography-mas spectrometry (GC-MS). Next, the analyses of the global effect of the miR-1914-5p in our cellular model, will be performed by the shotgun proteomics strategy. To effective functional evaluadated the proteomic analyses relevant and selected proteins will be closer investigated by Western blot, biochemistry analyses and gene silencing or its protein overexpression in cell culture. (AU)