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ANALYSIS OF INFLAMMASOME INDUCED BY URATE MONOSSODIUM IN MONOCYTES FROM PREGNANT WOMEN WITH PREECLAMPSIA

Grant number: 12/21287-3
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2013
Effective date (End): February 28, 2015
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal researcher:Maria Terezinha Serrão Peraçoli
Grantee:Mariana Leticia Matias
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated scholarship(s):14/00693-9 - Effect of hyperuricemia in the plasma of pregnant women with preeclampsia on autophagy in mononuclear lymphoid cells, BE.EP.MS

Abstract

Preeclampsia (PE) is a clinical complication of pregnancy characterized by hypertension and proteinuria, identified after the 20th week of gestation. This pathology is associated with hyperuricemia, elevated serum levels of inflammatory cytokines, leukocyte activation and oxidative stress. Higher levels of uric acid in plasma of pregnant women with PE have been considered not only as a marker of severity, but represent a direct contribution to the pathogenesis of PE. Uric acid crystals can activate an intracellular complex named inflammasome, a multi-protein complex, important for processing and release of inflammatory cytokines such as IL-1beta and IL-18. Nod-like receptors or NLR (protein containing nucleotide oligomerization domain) are cytosolic proteins present in cells of innate immunity that recognize pathogen-associated molecular patterns or endogenous cytoplasmic products and recruit other proteins, forming signaling complexes that promote inflammation. The present project aims to: a) to assess the endogenous or monosodium urate-induced activation state of monocytes from pregnant women with preeclampsia by NLRP1/NLRP3 inflammasome identification in these cells, and association of this complex with IL-1beta, IL-18 and TNF-alpha production by these cells, b) to correlate plasma levels of uric acid with inflammasome activation and inflammatory cytokine production by monocytes from women with PE. Forty six pregnant women being 23 normotensive and 23 with preeclampsia, matched for gestational age as well as 23 healthy non-pregnant women will be included in the study. Peripheral blood monocytes will be incubated at 37oC in constant atmosphere of 5% CO2 in the presence or absence of monosodium urate. The supernatant obtained after 18h of culture will be harvested and employed for IL-1beta, IL-18 and TNF-alpha determination by enzyme immunoassay (ELISA). The presence of inflammasome in these cells will be evaluated by gene expression of NLRP1/NLRP3, ASC, caspase-1 and IL-1beta after blood collection by quantitative real-time PCR (qPCR) (endogenous expression) or after in vitro activation with monosodium urate (stimulated expression).

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
MATIAS, Mariana Leticia. Análise de inflamassoma induzido por urato monossódico em monócitos de gestantes portadoras de pré-eclampsia. 2015. 98 f. Master's Dissertation - Universidade Estadual Paulista "Júlio de Mesquita Filho" Faculdade de Medicina..

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