Immunomodulatory action of propolis on monocytes of pregnant women with preeclampsia and on human umbilical vein endothelial cells (HUVEC)

• Grant number: 20/14639-7
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): April 01, 2022
• Effective date (End): July 31, 2024
• Field of knowledge: Biological Sciences - Pharmacology - Ethnopharmacology
• Principal Investigator: José Maurício Sforcin
• Grantee: Vanessa Rocha Ribeiro Vasques
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated research grant: 17/04138-8 - Attainment of chemical, analytical, biological, pharmacological and technological studies to fill the gaps on the development of Brazilian propolis sector, AP.TEM

Abstract

Literature data demonstrate that preeclampsia (PE) is an important pregnancy syndrome, resulting from sterile inflammation. Endogenous triggering factors are known as damage-associated molecular patterns (DAMPs), such as Hsp70 and HMGB1. The release of DAMPs associated with oxidative stress, induced by monocyte activation, high production of pro-inflammatory cytokines, free radicals, adhesion molecules and decreased antioxidants, can cause endothelial changes, characteristics of this syndrome. Thus, the study of agents that can modulate the inflammatory state of these pregnant women needs further investigation. In this context, propolis is a bee product with numerous pharmacological properties, highlighting its immunomodulatory and anti-inflammatory action. The objective of this work is to analyze the action of propolis on sterile inflammation and endothelial dysfunction, through in vitro treatment of monocytes obtained from preeclamptic pregnant women and human umbilical vein endothelial cells (HUVEC). HUVEC will be cultured in the presence or absence of plasma from pregnant women with PE, HMGB1 and Hsp70, and monocytes obtained from 32 pregnant women, 16 with early-onset PE and 16 with late-onset PE will be treated in the presence or absence of nanoparticles containing propolis. The levels of HMGB1, Hsp70 and MCP-1 will be determined in the plasma of pregnant women by immunoenzymatic assays. The analysis of the expression of ICAM-1, VCAM-1, MCP-1, E-selectin, endothelin1, endoglobin, VEGFR1, VEGFR2 will be evaluated in HUVEC, while the ERK1/ 2 and NF-kB signaling pathways and the intracytoplasmic cytokines IL -1, IL-6, IL-8, IL-10, IL-12, TNF-alpha will be evaluated both in HUVEC and monocytes by flow cytometry. The dosage of TOS, TAS and HO-1 will be determined in the supernatant of the monocytes and sFlt1, sENG and VEGF in the supernatant of the HUVEC, through commercial kits. Biomarkers of endothelial dysfunction will be evaluated in HUVEC by immunofluorescence. The results will be analyzed using parametric or non-parametric tests with a significance level of 5%. The data obtained may culminate in a new therapeutic approach to improve maternal and fetal prognosis.