Modulatory effect of progesterone, vitamin D and silibinin on the inflammasomes in monocytes from pregnant women with preeclampsia

Author(s)

Mariana Leticia Matias

• Document type: Doctoral Thesis
• Press: Botucatu. 2019-03-18.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Medicina. Botucatu
• Defense date: 2019-02-28
• Advisor: Maria Terezinha Serrão Peraçoli

Abstract

Preeclampsia (PE) is a specific pregnancy syndrome, associated with exacerbated systemic inflammation, which could be dependent on the deficiency of regulatory factors, such as silibinin (SB), progesterone (PG), and vitamin D (VD), which are able to modulate this inflammatory response. The aim of this study was to evaluate the modulating effect of SB, PG and VD on the NLRP1 and NLRP3 inflammasomes and on the TLR4-MyD88-dependent signaling pathway in monocytes from pregnant women with PE and human monocyte cell line THP-1. Monocytes obtained from the peripheral blood of 20 pregnant women with PE, 20 normotensive pregnant women and THP-1 cells were cultured in the presence or absence of monosodium urate (MSU), hyaluronan (HA), SB, PG or VD for different periods: a) for 30 min for the determination of NF-κB phosphorylation and for the expression of the intra-cytoplasmic factors NF-κB and IκBα by flow cytometry, b) for 4 h for the gene expression of TLR4, MYD88, NFKB1, NLRP1, NLRP3, CASP1, IL1B, IL18, TNF and IL10, analyzed by qPCR and c) for 18h for the determination of IL-1β, IL-18, TNF-α and IL-10 cytokines concentration by ELISA and for expression of the surface receptor TLR4 by flow cytometry. Plasma levels of uric acid, PG and VD in preeclamptic and normotensive pregnant women were determined. The results were analyzed using parametric and non-parametric tests with a significance level of 5%. Plasma levels of PG and VD were significantly lower, whereas those of uric acid were higher in pregnant women with PE when compared to normotensive pregnant women. Monocytes from pregnant women with PE showed greater endogenous activation of the inflammatory genes (TLR4, MYD88, NFKB1, NLRP1, NLRP3, CASP1, IL1B, IL18 and TNF) and lower IL10 when compared to normotensive pregnant women. The expression of TLR4 and NF-κB was higher, whereas IκBα was lower in the group of pregnant women with PE than in normotensive pregnant women. The higher basal activity of phosphorylated NF-κB observed in monocytes of pregnant women with PE was lower after the treatment of these cells with SB, PG and VD. The stimulation with MSU and HA increased the expression of genes and proteins related to inflammation and decreased IL-10 in pregnant women with PE, whereas treatment with SB, PG and VD had an immunomodulatory effect, decreasing this expression. In PE group, MSU and HA stimuli increased NF-κB expression and decreased IκBα expression. On the other hand, treatment of monocytes with SB, PG and VD induced a decrease of TLR4 and NF-κB and increase of IκBα. THP-1 cells presented a similar immune response profile to monocytes from preeclamptic women after MSU and HA stimulation and SB, PG and VD treatment. The results suggest the participation of MSU and HA in the systemic inflammatory response in PE and that treatment with SB, PG or VD is able to modulate this inflammation, demonstrating the immunomodulatory role of these substances in the regulation of the inflammatory response in PE. (AU)

• FAPESP's process: 15/26147-3 - Modulatory effect of progesterone, vitamin D and silibinin on the inflammasomes in monocytes from pegnant women with peeclampsia
• Grantee: Mariana Leticia Matias
• Support Opportunities: Scholarships in Brazil - Doctorate