Navegando por Palavras-chave "ApoE-/- Knockout Mouse"

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    Biologia vascular: síntese e remodelamento de moléculas da matriz extracelular e de superfície de células endoteliais em artéria carótida de camundongos ApoE-/-.
    (Universidade Federal de São Paulo (UNIFESP), 2021) Russo, Thatiane Amaral [UNIFESP]; Regatieri, Juliana Luporini Dreyfuss [UNIFESP]; Universidade Federal de São Paulo
    The endothelium is composed of a monolayer of cells and their basemant membrane, which line the inside of the blood vessels and are responsible for maintaining vascular homeostasis. Under physiological conditions, it maintains vascular tone, laminar blood flow, coagulation process, cell proliferation and migration and control of the inflammatory response. Endothelial cells (ECs) respond to mechanical forces such as cell stretching and shear stress, influencing cellular behavior such as: morphology, migration, adhesion and angiogenesis, where many extracellular matrix (ECM) and cell surface components play a fundamental role. Endothelial dysfunction can cause cardiovascular diseases, such as hypertension, thrombosis and atherosclerosis, and these diseases can be modulated by the mechanical forces applied to the artery wall. This study aimed to investigate the remodeling of ECM and molecules associated with this remodeling in ECs submitted in vitro to two mechanical stimuli, cell stretching and shear stress, both in physiological and pathological conditions. After the stimuli, tests of cell behavior (adhesion, cell migration and formation of capillary structures), analysis of the biosynthesis of glycosaminoglycans by incorporating 35S, immunofluorescence, qPCR and qPCR array were performed. Both mechanical forces induced changes the ECs morphology, cell adhesion, migration and formation of capillary structures in culture were modulated positively in physiological conditions and negatively in pathological situations. The pathological mechanical forces applied to ECs induced a higher gene expression of syndecan-4, perlecan, versican, decorin, fibronectin, collagen III α1, connexin-43, VEGFA, TGF-β1 and TGF-β3. In addition, in vivo assays with ApoE-/- deficient mice were used as an animal model to study ECM remodeling in atherosclerotic disease. For this purpose the carotids of ApoE-/- mice were analyzed by high resolution ultrasound, demonstrating that there was an increase in the internal diameter and internal thickness (IMT) of these vessels, characterizing an artery dysfunction. Then these carotids were removed and subjected to analysis of gene expression by qPCR and qPCR array, immunofluorescence and histopathology. It was possible to observe a higher gene expression and a higher fluorescence intensity of syndecan-4, perlecan, versican and connexin-43 in carotids of ApoE-/- mice when compared to wild type control mice. The results of the qPCR array demonstrated higher expression of adhesion molecules, laminins and integrins and genes related to ECM proteolysis in carotids of ApoE-/- mice when compared to wild type control mice. This study helps to understand the remodeling and associated molecules of the ECM in in vitro and in vivo models of cardiovascular diseases, and how the endothelium can be affected by the mechanical forces of cell stretching and shear stress.