Efeito de exossomos do tecido adiposo de camundongos obesos na polarização de macrófagos

Abstract

The worldwide growth in the number of obesity cases is a matter of concern, mainly because this condition is a risk factor for the development of other comorbidities resulting from the chronic inflammatory process in the adipose tissue. This tissue can communicate through hormones, inflammatory mediators, and recent studies describe the importance of extracellular vesicles in this process, with these being able to interact even with the immune system. Of the immune cells, macrophages are the ones that are present in greater numbers in the adipose tissue, and it has been reported that, in the context of obesity, they are directed towards a pro-inflammatory profile. Thus, the objective of this work is to evaluate how exosomes from obese murine adipose tissue interact with macrophages. For this, C57BL/6 animals were submitted to the obesity model, the adipose tissue gonadal fat was collected to obtain the vesicles, in addition to the collection of bone marrow from the control group for differentiation into macrophages. The standardization of the obtainment of exosomes by centrifugation and ultracentrifugation was carried out, with the size being evaluated by the DLS and the amount of protein by the BCA kit, with the data found so far being corroborated by those found in the literature. The working concentration of exosomes (25 and 50 µg/mL) was established and the standardization of obtaining bone marrow-derived macrophages was carried out. The efficiency of the macrophage differentiation method was verified by analyzing the expression of F4/80 by flow cytometry. Then, the vesicles were characterized in terms of size, morphology, presence of specific markers and amount of proteins. Primary culture macrophages were then treated with exosomes and evaluated for mediators produced and polarization profile