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Effects of supernatant from placental explants treated with vitamin D on oxidative stress, inflammation and nitric oxide availability in endothelial cells

Grant number: 19/00234-8
Support type:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): May 06, 2019
Effective date (End): August 05, 2019
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal researcher:Maria Terezinha Serrão Peraçoli
Grantee:Priscila Rezeck Nunes
Supervisor abroad: Mattias Carlstrom
Home Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Research place: Karolinska Institutet, Sweden  
Associated to the scholarship:16/23452-2 - Immunomodulatory effect of vitamin D on the activation of inflammasomes in placental tissue from pregnant women with pre-eclampsia, BP.DR

Abstract

Preeclampsia (PE) is an exclusive syndrome of human gestation that is beginning withlocal placental dysfunction and develops into a disease identified by maternal hypertension and/or proteinuria. Nitric oxide (NO) is a biological mediator able to participate in the process related to gestation, representing the main vasodilator in theplacenta.Disturbances in NO system is responsible for the vascular endothelialdysfunction present in PE and together with excess of reactive oxygen species (ROS)production, may explain the poor placental perfusion in PE. The aim of this study is toevaluate whether human umbilical vein endothelial cells (HUVEC) cultured withsupernatant from placental explants of normotensive (NT) and preeclamptic pregnantwomen with Vitamin D decrease oxidative stress and increase the bioavailability of NO.Human umbilical vein endothelial cell line will be cultured until reaching 80% confluenceand then incubate for 4 and 24 hours in medium with 20% (v/v) of supernatant fromplacental explants cultured with or without hydrogen peroxide (H2O2) and vitamin D (VD).The cells and supernatant will be used to perform the assays. Electron paramagneticresonance (EPR) and high performance liquid chromatography (HPLC) system will beused to assess NO and its oxidized metabolites nitrate and nitrite, but also specific ROS.Determination of TBARS (Thiobarbituric acid reactive substances) will be employ toevaluate ROS-mediated lipidic peroxidation, as well as asymmetric dimethylarginine(ADMA). Cytokines including IL-1², IL-18, TNF-±, IL-10 and caspase-1 will be evaluatedby Multiplex ELISA assay. Statistical analysis (ANOVA or t-test) will be performedemploying parametric or nonparametric tests, with p value of <0.05 consideredsignificant.

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