Endothelial cells behavior submitted to an in vitro arterial hypertension model

Author(s)

Thais Silva Pinto

• Document type: Master's Dissertation
• Press: Botucatu. 2019-04-29.
• Institution: Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu
• Defense date: 2019-02-22
• Advisor: Willian Fernando Zambuzzi

Abstract

Shear-stress changes are associated with a repertory of signaling cascade, modulating vascular phenotype. As shear stress-related tensional forces might be associated with pathophysiological susceptibility, a more comprehensive molecular map needs to be addressed. Thus, we subjected human umbilical vein endothelial cells (HUVECs) to a circuit of different tensional forces in vitro considering the following three groups: one in a physiological blood flow shear-stress condition (named Normo), another in which these cells followed to a hypertensive blood flow shear-stress (named Hyper), and finally one that these hyper-stressed cells were returned to Normo condition (named Return). The samples were properly collected to allow different methodologies analysis. Our data showed a pivotal involvement of c-Src on driving the mechanotransduction cascade by modulating signaling related with adhesion, survival (PI3K/Akt) and proliferative phenotype. Moreover, c-Src seems to develop important role during Extracellular Matrix (ECM) remodeling, which showed significative changes. Additionally, proteomic analysis showed strong involvement of Heat Shock Protein 70 (HSP70) in the hypertensive-stressed cells; it being significantly decreased in Return phenotype. This result prompted us to investigate 20S proteasome as an intracellular proteolytic alternative to promote the turnover of those proteins. Surprisingly, our data reveled significant over expression of sets of proteasome subunit α-type (PSMA) and proteasome subunit β-type (PSMB) genes (encoding proteins related with catalytic core of proteasome). Besides, constituents of classical intracellular pathways - for example Wnt/βCatenina, Bax and BCl-2, including genes involved with energetic metabolism, also suffered strong modulation on different model pressure. Altogether, our data reported that an in vitro hypertension model can bring a lot of insights about the mechanosensitivity in endothelial cells. Since shear patterns are associated with pathophysiologic changes, as well atherosclerosis and hypertension, these results paved new roads to understand the molecular mechanism in mechanotransduction driving in endothelium, and if drugable, many of these targets should be considered for future pre-clinical research. (AU)

• FAPESP's process: 17/18349-0 - Endothelial cell behavior under arterial hypertension in vitro and its impact on osteogenesis
• Grantee: Thaís Silva Pinto
• Support Opportunities: Scholarships in Brazil - Master