Evaluation of pharmacological and molecular inhibition of NLRP3 inflammasome and its consequences in an in vitro model of Preeclampsia

Abstract

Preeclampsia (PE) is a specific syndrome of a human pregnancy, being one of the main causes of maternal death in Brazil. The pathophysiology of PE has not been fully clarified, but it is known that placental ischemia is of fundamental importance to the process, since the release of products resulting from malperfusion in maternal circulation can lead to systemic endothelial dysfunction. A persistent endothelial inflammation stimulates the secretion of various inflammatory mediators, which triggers an important process in inflammatory conditions due to the activation of various signalization patterns. Considering that the causes related to endothelial dysfunction, oxidative stress, and inflammation in PE still remain a challenge for clinical practice, the use of drugs and the use of molecular approaches related to the inhibition of NLRP3 inflammasome could be a good choice for future treatments. The objective of this project is: to assess the pharmacological and molecular action of NLRP3 inflammation inhibitors in an in vitro model of PE and the relationships with biomarkers of endothelial dysfunction, oxidative stress and inflammation. To achieve this objective, the following experiments will be carried out: 1- assess the effect of plasma from pregnant women with PE and normotensive (NT) pregnant women in the activation of the NLRP3 inflammassome via NF-ºB in endothelial cell line; 2- comparison of the activation of the inflammation in primary endothelial cells of pregnant women with PE and NT subjected to treatment with pharmacological inhibitors and small interfering RNA; 3- validate the effect the pharmacological and molecular inhibition on the function of endothelial cells incubated with plasma through tests of tube formation, permeability, markers of endothelial dysfunction, oxidative stress, autophagy and pyroptose; 4- observe the effect of two inhibitors of NLRP3 inflammassome on contraction and relaxation of vessels from pregnant women by vascular reativity test. The results will be validated by parametric or non-parametric tests, depending on the variability, considering significance level <0.05. Studies on endothelial dysfunction and its relationship with inflammation in PE still remain an objective to be further explored and elucidated, and the molecular mechanisms associated with the interactions between inflammation and endothelial dysfunction can be favorable to develop new treatments and more effective therapies. (AU)