Nuclear Factor Kappa B Activation Occurs in the Amnion Prior to Labour Onset and Modulates the Expression of Numerous Labour Associated Genes

Background: Prior to the onset of human labour there is an increase in the synthesis of prostaglandins, cytokines and chemokines in the fetal membranes, particular the amnion. This is associated with activation of the transcription factor nuclear factor kappa B (NFkB). In this study we characterised the level of NFkB activity in amnion epithelial cells as a measure of amnion activation in samples collected from women undergoing caesarean section at 39 weeks gestation prior to the onset of labour. Methodology/Principal Findings: We found that a proportion of women exhibit low or moderate NFkB activity while other women exhibit high levels of NFkB activity (n = 12). This activation process does not appear to involve classical pathways of NFkB activation but rather is correlated with an increase in nuclear p65-Rel-B dimers. To identify the full range of genes upregulated in association with amnion activation, microarray analysis was performed on carefully characterised nonactivated amnion (n = 3) samples and compared to activated samples (n = 3). A total of 919 genes were upregulated in response to amnion activation including numerous inflammatory genes such cyclooxygenase-2 (COX-2, 44-fold), interleukin 8 (IL-8, 6-fold), IL-1 receptor accessory protein (IL-1RAP, 4.5-fold), thrombospondin 1 (TSP-1, 3-fold) and, unexpectedly, oxytocin receptor (OTR, 24-fold). Ingenuity Pathway Analysis of the microarray data reveal the two main gene networks activated concurrently with amnion activation are i) cell death, cancer and morphology and ii) cell cycle, embryoni

Leukocyte density and proinflammatory mediator expression in regional human fetal membranes and decidua before and during labor at term

OBJECTIVES: The region of fetal membranes overlying the cervix, known as the zone of altered morphology (ZAM), is considered to be the principle site of membrane inflammatory activity and extracellular matrix remodelling. We wished to quantify the relative contribution of each area of fetal membranes to the inflammatory process of parturition. Specifically, we aimed to quantify and compare (1) leukocyte densities in three regions of fetal membranes and decidua before and during spontaneous labor at term, and (2) mRNA expression of interleukin (IL)-1 beta, IL-6, IL-8, cyclo-oxygenase type 1 (COX-1), and COX-2 in three regions of fetal membranes and decidua before and during spontaneous labor at term. METHODS: Biopsies of fetal membranes and decidua were obtained from pregnant women delivered by cesarean section at term both before and during spontaneous labor (n = 8 both groups). Fetal membranes were sampled from three areas, the ZAM, midzone (MZ), and periplacental (PP) regions. Leukocytes were identified by immunohistochemistry and their density quantified. Inflammatory mediator expression was quantified using TaqMan technology (Applied Biosystems, Foster City, CA). RESULTS: There was a significantly greater density of leukocytes in (1) the PP region of membranes compared with the ZAM, and (2) the decidua compared with amnion, amniotic connective tissue, and chorion. IL-1 beta, IL-6, and IL-8 mRNA expression was significantly greater in all regions following spontaneous labor compared with nonlaboring tissues. There were no regional differences in cytokine expression within the fetal membranes. Choriodecidua expressed significantly more IL-1 beta mRNA than amnion. Amnion expressed more COX-2 mRNA than choriodecidua. CONCLUSIONS: All regions of fetal membranes and decidua contribute to the inflammatory process of human parturition; however, their relative contributions differ in magnitude. Although the ZAM may be specifically important for membrane rupture, it does not appear to play a key or exclusive role in the other inflammatory processes of parturition. When studying fetal membranes, it is relevant to identify and define the area sampled for consistency and comparison with other studies

Labor-associated regulation of prostaglandin E and F synthesis and action in the ovine amnion and cervix

OBJECTIVE: Prostaglandins (PGs) are key regulators of cervical dilatation and membrane breakdown at the onset of labor. PG synthase and receptor expression has been previously documented in uterine tissues; however, mechanisms governing the changes occurring in the cervix and amnion are less well established. The aim of the current study was to determine the level of expression of PG synthetic enzymes and receptors in these tissues in association with induced labor in sheep