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 Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: May 06, 2019
• End date: August 05, 2019
• Field of knowledge: Health Sciences - Medicine - Maternal and Child Health
• Principal Investigator: Maria Terezinha Serrão Peraçoli
• Grantee: Priscila Rezeck Nunes
• Supervisor: Mattias Carlstrom
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: 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.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

NUNES, P. R.; GOMES, V. J.; SANDRIM, V. C.; PERACOLI, J. C.; PERACOLI, M. T. S.; CARLSTROM, M.. Effects of vitamin D-induced supernatant of placental explants from preeclamptic women on oxidative stress and nitric oxide bioavailability in human umbilical vein endothelial cells. Brazilian Journal of Medical and Biological Research, v. 54, n. 8, 2021. (16/23452-2, 19/00234-8)