Identification of metabolic pathways related to endothelial dysfunction of pre-eclampsia in vitro model

Abstract

Preeclampsia (PE) is a specific syndrome of human pregnancy and the clinical parameters that define it are well understood. However, given the severity for the mother and fetus and the confirmation of the diagnosis only after the twentieth week of gestation, many studies have evaluated the use of biomarkers, such as the ratio between sFlt-1 (soluble vascular endothelial growth factor receptor) and PlGF (placental growth factor) in the prediction of PE. These biomarkers are two pro (PlGF) and anti-angiogenic (sFlt-1) molecules released by the placenta during pregnancy. In addition to the angiogenic balance, another important mechanism in pregnancies complicated by PE is the persistent inflammation in the endothelium, which stimulates the secretion of several inflammatory mediators, responsible for the activation of different signaling patterns. An important inflammatory mediator is uric acid, which in the form of crystals (monosodium urate - MSU) contributes to the pathogenesis of PE through its inflammatory effects. Considering the angiogenic imbalance and the exacerbated inflammatory state in pregnant women with PE, the objective of the project will be to evaluate the activation of the metabolic pathways involved in an in vitro model (endothelial cell culture - HUVEC) of PE and the effect of incubation with sFLT-1/ PlGF and MSU, to confirm markers of endothelial dysfunction and inflammation, which may aid in the identification of high-risk pregnancies complicated by PE. Metabolic profiles will be determined using the Seahorse Analyzer XE24 (Agilent), evaluating the rate of oxygen consumption and the rate of extracellular acidification in the presence of modulators of specific metabolic pathways (glycolysis, beta-oxidation, mitochondrial respiration). Univariate statistical analysis will be performed using GraphPad Prisma (version 7.0). Results will be expressed as mean and standard error of the mean (SEM). The results will be evaluated by parametric or non-parametric tests, depending on the variability, with a significance level of p<0.05 being adopted. Studies on endothelial dysfunction and its relationship with inflammation in PE still remain an objective to be further explored. Thus, elucidating the molecular and metabolic mechanisms associated with the interactions between inflammation and endothelial dysfunction may be favorable to the development of new treatments and more effective therapies. The analysis of metabolic parameters may provide information not identified by traditional diagnoses and with potential future biomarkers. (AU)