scholarly journals KLF5 regulates infection- and inflammation-induced pro-labour mediators in human myometrium

Reproduction ◽  
2015 ◽  
Vol 149 (5) ◽  
pp. 413-424 ◽  
Author(s):  
Martha Lappas
Keyword(s):  
Mrna Expression ◽  
Interleukin 1 ◽  
Poly I:C ◽  
Human Myometrium ◽  
Viral Dsrna ◽  
Myometrial Cells ◽  
Human Labour ◽  
Foetal Membranes ◽  
Term Labour ◽  
Klf5 Expression

The transcription factor Kruppel-like factor 5 (KLF5) has been shown to associate with nuclear factor kappa B (NFκB) to regulate genes involved in inflammation. However, there are no studies on the expression and regulation of KLF5 in the processes of human labour and delivery. Thus, the aims of this study were to determine the effect of i) human labour on KLF5 expression in both foetal membranes and myometrium; ii) the pro-inflammatory cytokine interleukin 1 beta (IL1β), bacterial product flagellin and the viral dsRNA analogue poly(I:C) on KLF5 expression and iii) KLF5 knockdown by siRNA in human myometrial primary cells on pro-inflammatory and pro-labour mediators. In foetal membranes, there was no effect of term or preterm labour on KLF5 expression. In myometrium, the term labour was associated with an increase in nuclear KLF5 protein expression. Moreover, KLF5 expression was also increased in myometrial cells treated with IL1β, flagellin or poly(IC), likely factors contributing to preterm birth. KLF5 silencing in myometrial cells significantly decreased IL1β-induced cytokine expression (IL6 and IL8 mRNA expression and release), COX2 mRNA expression, and subsequent release of prostaglandins PGE2 and PGF2α. KLF5 silencing also significantly reduced flagellin- and poly(I:C)-induced IL6 and IL8 mRNA expression. Lastly, IL1β-, flagellin- and poly(I:C)-stimulated NFκB transcriptional activity was significantly suppressed in KLF5-knockout myometrial cells. In conclusion, this study describes novel data in which KLF5 is increased in labouring myometrium, and KLF5 silencing decreased inflammation- and infection-induced pro-labour mediators.

scholarly journals RAF1 is increased in labouring myometrium and modulates inflammation-induced pro-labour mediators

Reproduction ◽  
2016 ◽  
Vol 151 (4) ◽  
pp. 411-420 ◽  
Author(s):  
Martha Lappas
Keyword(s):  
Protein Expression ◽  
Interleukin 1 ◽  
Signalling Pathway ◽  
Mrna Abundance ◽  
Mrna Levels ◽  
Myometrial Cells ◽  
Human Labour ◽  
Prostaglandin Endoperoxide Synthase ◽  
Sirna Knockdown ◽  
Term Labour

Inflammation plays a central role in the terminal process of human labour and delivery, including myometrial contractions. RAF1 proto-oncogene serine/threonine-protein kinase (RAF1) can activate ERK (official gene symbolMAPK1) and/or nuclear factor-kappa B (NF-κB) to regulate genes involved in inflammation. There are, however, no studies on the role of RAF1 in the processes of human labour and delivery. Thus, the aims of this study were to determine the effect of i) human labour and pro-inflammatory cytokines interleukin 1 beta (IL1B) and tumour necrosis factor (TNF) alpha on RAF1 protein expression in myometrium and ii) siRNA knockdown ofRAF1on pro-inflammatory and pro-labour mediators in human myometrial primary cells. Term labour was associated with an increase in RAF1 protein expression. Furthermore, RAF1 protein expression was increased in myometrial cells treated with IL1B and TNF, two likely factors contributing to preterm birth. Knockdown ofRAF1by siRNA in primary myometrial cells significantly decreased IL1B- and TNF-inducedIL1A, IL1B, IL6,(C-X-C motif) ligand 8 (CXCL8)and chemokine (C-C motif) ligand 2 (CCL2) mRNA abundance and IL6, IL8 and CCL2; prostaglandin-endoperoxide synthase 2 (PTGS2) mRNA levels and prostaglandin PGF2αrelease; and NF-κB activation. Furthermore,RAF1knockdown was associated with decreased activation of ERK in the presence of IL1B but not TNF. Concordantly, the ERK inhibitor U0126 significantly decreased IL1B-inducedIL6,CXCL8,CCL2andPTGS2mRNA abundance; IL6, CXCL8, CCL2 and PGF2αrelease; and NF-κB activation. In conclusion, IL1B induces the expression and secretion of pro-labour mediators through the RAF1–MAPK1–NF-κB signalling pathway. TNF, on the other hand, regulates pro-labour mediators through the RAF1–NF-κB signalling pathway via an MAPK1-independent mechanism.

scholarly journals IRF5 is increased in labouring myometrium and regulates pro-labour mediators

Reproduction ◽  
2018 ◽  
Vol 156 (3) ◽  
pp. 207-218
Author(s):  
Ratana Lim ◽  
Gillian Barker ◽  
Martha Lappas
Keyword(s):  
Preterm Birth ◽  
Inflammatory Response ◽  
Inflammatory Cytokines ◽  
Treatment Options ◽  
Adult Life ◽  
Human Myometrium ◽  
Myometrial Cells ◽  
Associated Proteins ◽  
Term Labour ◽  
Pro Inflammatory Cytokines

Preterm birth continues to be the leading cause of neonatal mortality and morbidities that can extend into adult life. Few treatment options stem from our incomplete understanding of the mechanisms of human labour and delivery. Activation of the inflammatory response in gestational tissues by inflammation and/or infection leads to the production of pro-inflammatory and pro-labour mediators, thus preterm birth. Interferon regulatory factor 5 (IRF5) has recently emerged as an important pro-inflammatory transcription factor involved in acute and chronic inflammation. The aims of this study were to determine the expression of IRF5 in human myometrium from labouring and non-labouring women, and whether IRF5 is involved in the genesis of pro-inflammatory and pro-labour mediators induced by pro-inflammatory cytokines or toll-like receptor (TLR) ligands. IRF5 mRNA and protein expression was significantly higher in human myometrium after spontaneous term labour, compared to non-labouring tissues. IRF5 mRNA expression was also significantly higher in primary myometrial cells treated with the pro-inflammatory cytokines IL1B or TNF. In primary myometrial cells, IRF5 knockdown by siRNA (siIRF5) was associated with significantly decreased expression and or secretion of pro-inflammatory cytokines (IL1A, IL6), chemokines (CXCL8, CCL2), adhesion molecules (ICAM1, VCAM1) and contraction-associated proteins PTGS2, PGF2α and PTGFR when in the presence of IL1B, TNF, fsl-1 (TLR2/6 ligand) or flagellin (TLR5 ligand). siIRF5-transfected cells also displayed decreased NF-κB RELA transcriptional activity in the presence of these preterm birth mediators. Our study suggests a novel role for IRF5 in the regulation of the inflammatory response in human myometrium.

scholarly journals DREAM Is Involved in the Genesis of Inflammation-Induced Prolabour Mediators in Human Myometrial and Amnion Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Priyanka Goradia ◽  
Ratana Lim ◽  
Martha Lappas
Keyword(s):  
Preterm Birth ◽  
Mrna Expression ◽  
Proinflammatory Cytokine ◽  
Transcriptional Activity ◽  
Regulatory Element ◽  
Poly I:C ◽  
Fetal Membranes ◽  
Perinatal Morbidity ◽  
Viral Dsrna ◽  
Spontaneous Labour

Preterm birth is the primary cause of perinatal morbidity and mortality worldwide. Inflammation induces a cascade of events leading to preterm birth by activating nuclear factor-κB (NF-κB). In nongestational tissues, downstream regulatory element antagonist modulator (DREAM) regulates NF-κB activity. Our aims were to analyse DREAM expression in myometrium and fetal membranes obtained at term and preterm and to determine the effect of DREAM inhibition on prolabour mediators in primary myometrial and amnion cells. DREAM mRNA expression was significantly higher in fetal membranes obtained after spontaneous labour compared to nonlabour and in amnion from women with histological preterm chorioamnionitis when compared to amnion from women without chorioamnionitis. In primary myometrial and amnion cells, the effect of DREAM silencing by siRNA was a significant decrease in the expression of proinflammatory cytokine IL-6, the chemokines IL-8 and MCP-1, the adhesion molecule ICAM-1, MMP-9 mRNA expression and activity, and NF-κB transcriptional activity when stimulated with the proinflammatory cytokine IL-1β, the bacterial products fsl-1 or flagellin, or the viral dsRNA analogue poly(I:C). These data suggest that, in states of heightened inflammation, DREAM mRNA expression is increased and that, in myometrial and amnion cells, DREAM regulates proinflammatory and prolabour mediators which may be mediated via NF-κB.

scholarly journals The Onset of Labor Alters Corticotropin-Releasing Hormone Type 1 Receptor Variant Expression in Human Myometrium: Putative Role of Interleukin-1β

Endocrinology ◽  
2007 ◽  
Vol 148 (7) ◽  
pp. 3205-3213 ◽  
Author(s):  
Danijela Markovic ◽  
Manu Vatish ◽  
Mei Gu ◽  
Donna Slater ◽  
Rob Newton ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Nuclear Translocation ◽  
Fold Increase ◽  
Nuclear Factor Κb ◽  
Prolonged Treatment ◽  
Human Myometrium ◽  
Mrna Levels ◽  
Myometrial Cells ◽  
Important Regulator ◽  
R1 Gene

CRH targets the human myometrium during pregnancy. The efficiency of CRH actions is determined by expression of functional receptors (CRH-R), which are dynamically regulated. Studies in myometrial tissue biopsies using quantitative RT-PCR demonstrated that the onset of labor, term or preterm, is associated with a significant 2- to 3-fold increase in CRH-R1 mRNA levels. Detailed analysis of myometrial CRH-R1 mRNA variants showed a decline of the pro-CRH-R1 mRNA encoding the CRH-R1β variant during labor and increased mRNA levels of CRH-R1d mRNA. Studies in myometrial cells identified IL-1β as an important regulator of myometrial CRH-R1 gene expression because prolonged treatment of myometrial cells with IL-1β (1 ng/ml) for 18 h induced expression of CRH-R1 mRNA levels by 1.5- to 2-fold but significantly attenuated CRH-R1β mRNA expression by 70%. In contrast, IL-1β had no effect on CRH-R1d mRNA expression. Studies using specific inhibitors suggest that ERK1/2, p38 MAPK, and downstream nuclear translocation of nuclear factor-κB mediate IL-1β effects on myometrial CRH-R1 gene. However, the increased CRH-R1 mRNA expression was associated with a dampening of the receptor efficacy to activate the adenylyl cyclase/cAMP signaling cascade. Thus, our findings suggest that IL-1β is an important regulator of CRH-R1 expression and functional activity, and this interaction might play a role in the transition of the uterus from quiescence to active contractions necessary for the onset of parturition.

433. Identifying novel biomarkers predictive of impending human labour

2008 ◽  
Vol 20 (9) ◽  
pp. 113
Author(s):  
Y. J. Heng ◽  
M. K. W. Di Quinzio ◽  
M. Permezel ◽  
G. E. Rice ◽  
H. M. Georgiou
Keyword(s):  
Predictive Value ◽  
Interleukin 1 ◽  
Fetal Membranes ◽  
Vaginal Fluid ◽  
Protease Inhibition ◽  
Fatty Acid Binding ◽  
Spontaneous Labour ◽  
Human Labour ◽  
Anti Inflammatory ◽  
Term Labour

Human labour is characterised by structural remodelling of the cervix and overlying fetal membranes, myometrial activation and parturition. We hypothesise that temporal biochemical alterations of the cervix and supracervical fetal membranes associated with impending labour may be reflected in the cervico-vaginal fluid (CVF). 2D PAGE proteomic analysis performed on serial CVF samples collected from women (n = 9) during late pregnancy and in spontaneous labour demonstrated 9 significantly altered protein spots (P < 0.05) in association with term labour. Seven different proteins were identified using electrospray ion-trap mass spectrometry (interleukin-1 receptor antagonist (IL-1ra), cystatin-A, glutathione S-transferase P, peroxiredoxin-2, thioredoxin, Cu,Zn superoxide dismutase, and epidermal fatty-acid binding protein). These proteins are involved in anti-inflammatory activity, protease inhibition, and oxidative stress defence. Validation of these potential biomarkers using ELISA is currently underway. Findings for the anti-inflammatory cytokine, IL-1ra are discussed. CVF was collected weekly from 106 women at 36 weeks' gestation up to and including spontaneous labour. The concentration of IL-1ra was 4-fold lower during labour compared with 15–21 and 22–28 days from labour (P < 0.05) and was also significantly lower (P < 0.05) at 0–7 days compared with 15–21 days before labour. After subdividing the women, the concentration of IL-1ra at 8–14 and 15–21 days before labour was 6-fold lower in women who had prelabour rupture of membranes at term followed by regular contractions compared with women who had spontaneous labour with intact membranes. Receiver-operator characteristic curve analysis indicated that IL-1ra best predicted term labour within 3 days of sampling with a cut-off value of 0.4 m g/mL (sensitivity 50.7%, specificity 72.2%, positive predictive value 36.1%, negative predictive value 82.6%). This IL-1ra validation study suggests that the decrease in IL-1ra in CVF during spontaneous term labour may be associated with proinflammatory-mediated remodelling of the fetal membranes leading to their rupture. (1) Di Quinzio et al. 2008, J. Proteome Res, 7:1916–1921. (2) Heng YJ et al. 2008, Am J Obstet Gynecol, In Press.

scholarly journals TBX2, a Novel Regulator of Labour

Medicina ◽  
2021 ◽  
Vol 57 (6) ◽  
pp. 515
Author(s):  
Febilla Fernando ◽  
Geertruda J.M. Veenboer ◽  
Martijn A. Oudijk ◽  
Marlies A.M. Kampman ◽  
Karst Y. Heida ◽  
...  
Keyword(s):  
Preterm Labour ◽  
Therapeutic Interventions ◽  
Human Myometrium ◽  
Mrna Levels ◽  
Myometrial Cells ◽  
Cytokines And Chemokines ◽  
Term Labour ◽  
Upstream Regulators

Background and Objectives: Therapeutic interventions targeting molecular factors involved in the transition from uterine quiescence to overt labour are not substantially reducing the rate of spontaneous preterm labour. The identification of novel rational therapeutic targets are essential to prevent the most common cause of neonatal mortality. Based on our previous work showing that Tbx2 (T-Box transcription factor 2) is a putative upstream regulator preceding progesterone withdrawal in mouse myometrium, we now investigate the role of TBX2 in human myometrium. Materials and Methods: RNA microarray analysis of (A) preterm human myometrium samples and (B) myometrial cells overexpressing TBX2 in vitro, combined with subsequent analysis of the two publicly available datasets of (C) Chan et al. and (D) Sharp et al. The effect of TBX2 overexpression on cytokines/chemokines secreted to the myometrium cell culture medium were determined by Luminex assay. Results: Analysis shows that overexpression of TBX2 in myometrial cells results in downregulation of TNFα- and interferon signalling. This downregulation is consistent with the decreased expression of cytokines and chemokines of which a subset has been previously associated with the inflammatory pathways relevant for human labour. In contrast, CXCL5 (C-X-C motif chemokine ligand 5), CCL21 and IL-6 (Interleukin 6), previously reported in relation to parturition, do not seem to be under TBX2 control. The combined bioinformatical analysis of the four mRNA datasets identifies a subset of upstream regulators common to both preterm and term labour under control of TBX2. Surprisingly, TBX2 mRNA levels are increased in preterm contractile myometrium. Conclusions: We identified a subset of upstream regulators common to both preterm and term labour that are activated in labour and repressed by TBX2. The increased TBX2 mRNA expression in myometrium collected during a preterm caesarean section while in spontaneous preterm labour compared to tissue harvested during iatrogenic preterm delivery does not fit the bioinformatical model. We can only explain this by speculating that the in vivo activity of TBX2 in human myometrium depends not only on the TBX2 expression levels but also on levels of the accessory proteins necessary for TBX2 activity.

scholarly journals PKA and AKIP1 interact to mediate cAMP-driven COX-2 expression: A potentially pivotal interaction in preterm and term labour

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252720
Author(s):  
Angela Yulia ◽  
Natasha Singh ◽  
Alice J. Varley ◽  
Kaiyu Lei ◽  
Danijela Markovic ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Human Myometrium ◽  
Interacting Protein ◽  
Protein Levels ◽  
Early Labour ◽  
Myometrial Contractility ◽  
Cox 2 ◽  
Nuclear Levels ◽  
Term Labour

Previously, we showed that cAMP increased COX-2 expression in myometrial cells via MAPK. Here, we have extended these observations, using primary myometrial cell cultures to show that the cAMP agonist, forskolin, enhances IL-1β-driven COX-2 expression. We then explored the role of A-kinase interacting protein (AKIP1), which modulates the effect of PKA on p65 activation. AKIP1 knockdown reversed the effect of forskolin, such that its addition inhibited IL-1β-induced COX-2 mRNA expression and reduced the IL-1β-induced increase in nuclear levels of p65 and c-jun. Forskolin alone and with IL-1β increased IκBα mRNA expression suggesting that in the context of inflammation and in the presence of AKIP1, cAMP enhances p65 activation. AKIP1 knockdown reversed these changes. Interestingly, AKIP1 knockdown had minimal effect on the ability of forskolin to repress either basal OTR expression or IL-1β-stimulated OTR mRNA expression. AKIP1 was up-regulated by IL-1β, but not stretch and was repressed by cAMP. The mRNA expression of AKIP1 increased in early labour in tandem with an increase in COX-2 mRNA and protein. AKIP1 protein levels were also increased with inflammation and stretch-induced preterm labour. Our results identify a second important cAMP effector-switch occurring at term in human myometrium and suggest that a hitherto unrecognized interaction may exist between AKIP1, NFκB and AP-1. These data add to the proposition that cAMP acts as a key regulator of human myometrial contractility.

scholarly journals Inhibition of GPR91 Reduces Inflammatory Mediators Involved in Active Labor in Myometrium

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Ratana Lim ◽  
Martha Lappas
Keyword(s):  
Mrna Expression ◽  
Preterm Labor ◽  
Statistical Significance ◽  
Collagen Gel ◽  
Human Myometrium ◽  
Myometrial Cells ◽  
Gm Csf ◽  
Proinflammatory Mediators ◽  
Collagen Gel Contraction ◽  
Associated Proteins

Problem. Some G-protein-coupled receptors (GPCRs) are regulators of inflammation, yet the role of the GPRC, GPR91, is unknown in human myometrium during the processes of human labor and delivery, a major inflammatory event. Method of Study. GPR91 mRNA expression was assessed using RT-qPCR in myometrium obtained from women at term Caesarean section in the absence of labor and during active spontaneous labor and in a mouse model of inflammation-induced preterm labor. Human primary myometrial cells were used to determine the effect of proinflammatory mediators on GPR91 and the effect of GPR91 siRNA on prolabor mediators. Statistical significance was ascribed to a P<0.05. Results. GPR91 mRNA expression was significantly higher in myometrium from women during term spontaneous labor compared to no labor. Likewise, in mice, GPR91 mRNA expression was significantly upregulated in myometrium during inflammation-induced preterm labor compared to preterm no labor. In myometrial cells, IL1B and TNF significantly increased GPR91 mRNA expression. Knockdown of GPR91 by siRNA in myometrial cells significantly suppressed the secretion and/or expression of IL1B- and TNF-induced proinflammatory cytokines (GM-CSF, IL1A, IL1B, and IL6) and chemokines (CXCL8 and CCL2), myometrial contractility (expression of the contraction-associated proteins PTGFR and CX43, secretion of the uterotonic PGF2α, and in situ collagen gel contraction), and the transcription factor NF-κB. Conclusion. Our findings demonstrate that GPR91 is involved in the genesis of proinflammatory and prolabor mediators induced by IL1B or TNF and collectively suggest that GPR91 may contribute to augmentation of the labor processes.

scholarly journals PARK7 regulates inflammation-induced pro-labour mediators in myometrial and amnion cells

Reproduction ◽  
2018 ◽  
Vol 155 (2) ◽  
pp. 207-218 ◽  
Author(s):  
Ratana Lim ◽  
Gillian Barker ◽  
Martha Lappas
Keyword(s):  
Preterm Birth ◽  
Poly I:C ◽  
Fetal Membranes ◽  
Uterine Contractility ◽  
Spontaneous Preterm Birth ◽  
Induced Expression ◽  
Neonatal Deaths ◽  
Myometrial Cells ◽  
Membrane Rupture ◽  
Term Labour

Preterm birth is a prevalent cause of neonatal deaths worldwide. Inflammation has been implicated in spontaneous preterm birth involved in the processes of uterine contractility and membrane rupture. Parkinson protein 7 (PARK7) has been found to play an inflammatory role in non-gestational tissues. The aims of this study were to determine the expression of PARK7 in myometrium and fetal membranes with respect to term labour onset and to elucidate the effect of PARK7 silencing in primary myometrium and amnion cells on pro-inflammatory and pro-labour mediators. PARK7 mRNA expression was higher in term myometrium and fetal membranes from women in labour compared to non-labouring samples and in amnion from preterm deliveries with chorioamnionitis. In human primary myometrial cells transfected with PARK7 siRNA (siPARK7), there was a significant decrease in IL1B, TNF, fsl-1 and poly(I:C)-induced expression of pro-inflammatory cytokine IL6, chemokines (CXCL8, CCL2), adhesion molecule ICAM1, prostaglandin PGF2α and its receptor PTGFR. Similarly, amnion cells transfected with siPARK7 displayed a decrease in IL1B-induced expression of IL6, CXCL8 and ICAM1. In myometrial cells transfected with siPARK7, there was a significant reduction of NF-κB RELA transcriptional activity when stimulated with fsl-1, flagellin and poly(I:C), but not with IL1B or TNF. Collectively, our novel data describe a role for PARK7 in regulating inflammation-induced pro-inflammatory and pro-labour mediators in human myometrial and amnion cells.

scholarly journals Is myometrial inflammation a cause or a consequence of term human labour?

2017 ◽  
Vol 235 (1) ◽  
pp. 69-83 ◽  
Author(s):  
Natasha Singh ◽  
Bronwen Herbert ◽  
Gavin R Sooranna ◽  
Nicolas M Orsi ◽  
Lydia Edey ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mrna Expression ◽  
Inflammatory Cell ◽  
Preterm Labour ◽  
Cytokine Mrna ◽  
Protein Levels ◽  
Cytokine Mrna Expression ◽  
Spontaneous Labour ◽  
Human Labour ◽  
Term Labour

Myometrial inflammation is thought to have a pivotal role in the onset of term and some forms of preterm labour. This is based on the comparison of samples taken from women undergoing term elective CS prior to the onset of labour with those taken from women in established labour. Consequently, it is not clear whether myometrial inflammation is a cause or a consequence of labour. Our objective is to test the hypothesis that myometrial inflammation is a consequence of the onset of labour. To test this hypothesis, we have obtained myometrial samples from women at various stages of pregnancy and spontaneous labour and studied the activation of the AP-1 (c-Jun) and NFκB (p65) systems, cytokine mRNA expression and protein levels and inflammatory cell infiltration and activation. We found that the activation of p65 declined from preterm to term not in labour samples and thereafter increased in early and established labour. Cytokine mRNA expression and protein levels increased in established labour only. Using flow cytometry of myometrial tissue, we found that the number of neutrophils did increase with the onset of labour, but on tissue section, these were seen to be intravascular and not infiltrating into the myometrium. These data suggest that myometrial inflammation is a consequence rather than a cause of term labour.

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