Expression of pentraxin-3 in chorioamniotic membranes from preterm pregnancies complicated by preterm labor or preterm premature rupture of membranes.

• Grant number: 11/17140-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2011
• End date: October 31, 2012
• Field of knowledge: Health Sciences - Medicine - Maternal and Child Health
• Principal Investigator: Márcia Guimarães da Silva
• Grantee: Amanda Manoel Della Coletta
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Introduction: Pentraxin 3 (PTX3) is an essential component of the innate immune response, and it acts as a pattern recognition receptor (PRR). It recognizes microbial products and opsonizes fungi, Gram-positive and Gram-negative bacteria, in addition to showing the capacity to activate the complement system. Few studies in the world literature have investigated PTX3 expression in gestational complications associated with microbial invasion of the amniotic cavity, and conflictive results have been reported. Objective: The present study aims at evaluating the gene and protein expression of PTX3 in chorioamniotic membranes of pregnancies complicated by the premature preterm rupture of membranes (PPROM) or preterm labor (PTL) in the presence of histologic chorioamnionitis. Patients and Methods: A cross-sectional study will be conducted. It will include 30 pregnant women in PTL (15 with histologic chorioamnionitis and 15 without) and 30 pregnant women with PPROM (15 with histologic chorioamnionitis and 15 without). All of these chorioamiotic membranes fragments have been properly stored in the Laboratory of Immunopathology of the Maternal-Fetal Relationship, Department of Pathology, Botucatu Medical School (FMB) - UNESP. The histologic chorioamnionitis diagnoses were confirmed by re-evaluation of slides filed at the slide collection of the Department of Pathology of FMB - UNESP. After thawing the stored fragments, 1cm2 of the membranes will be submitted to total RNA extraction. Following RNA extraction, the samples with concentrations between 0.02 and 0.2mg/mL of RNA will be submitted to cDNA collection for later use in quantifying PTX3 gene expression by the real-time PCR method utilizing the TaqMan® Gene Expression Assays System. Another fragment of the chorioamniotic membranes, adjacent to that used for gene expression, will be processed for protein analysis, which will be performed by the Western Blotting technique. Statistical analysis as well as the choice of tests for comparing PTX3 expression in the formed groups will be performed according to the presuppositions determined by the results, characteristics and behavior of the variables under study. Software SigmaStat 9.0 (Jandel Corporation) will be used, and the level of significance adopted for the tests will be of 5%.