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Regulation of phospholipase isozymes by nuclear factor-kappaB in human gestational tissues in vitro.
- Source :
-
The Journal of clinical endocrinology and metabolism [J Clin Endocrinol Metab] 2004 May; Vol. 89 (5), pp. 2365-72. - Publication Year :
- 2004
-
Abstract
- Phospholipid-derived mediators are implicated in the initiation and progression of human labor and delivery, particularly in relation to infection-induced preterm labor. We previously demonstrated that, in human intrauterine tissues, lipopolysaccharide (LPS)-stimulated nuclear factor-kappaB (NF-kappaB) DNA binding activity, and subsequent cytokine release can be suppressed by sulfasalazine (SASP) concentrations greater than 5 mM. The aim of this study was to elucidate the effect the SASP on secretory type II phospholipase A(2) (PLA(2)), cytosolic PLA(2) (cPLA(2)), cyclooxygenase (COX) isozymes, and subsequent prostaglandin F(2alpha) (PGF(2alpha)) production in human gestational tissues. Human placenta, amnion, and choriodecidua (n = 4-9 separate placentas) were incubated in the presence of SASP (0.1, 1, 5, and/or 10 mM) under either basal or LPS (10 microg/ml) conditions. After 6 h incubation, the tissues were collected and assayed for type II PLA(2) by ELISA and cPLA(2), COX-1, and COX-2 content by Western blotting. The incubation medium was collected and assayed for type II PLA(2) and 13,14-dihydro-15-keto PGF(2alpha) release by ELISA and PGF(2alpha) by RIA. Treatment of placenta, amnion, and choriodecidua with SASP concentrations greater than 5 mM significantly inhibited basal and/or LPS-stimulated type II PLA(2) content and release, 13,14-dihydro-15-keto PGF(2alpha) release, and cPLA(2) protein content (ANOVA, P < 0.05); however, no effect of SASP was observed on basal or LPS-stimulated COX-1 or COX-2 protein. Although no effect of SASP was observed on basal and LPS-stimulated PGF(2alpha) release from placenta and amnion, it significantly increased both basal and LPS-stimulated PGF(2alpha) release from choriodecidua. In addition, SASP concentrations of 5 mM or greater significantly suppressed NF-kappaB DNA binding activity. These data are consistent with the hypothesis that NF-kappaB regulates the expression and release of phospholipase isozymes.
- Subjects :
- Amnion enzymology
Amnion metabolism
Chorion enzymology
Chorion metabolism
Cyclooxygenase 1
Cyclooxygenase 2
Cytosol enzymology
DNA metabolism
Decidua enzymology
Decidua metabolism
Dinoprost metabolism
Electrophoresis
Enzyme-Linked Immunosorbent Assay
Extraembryonic Membranes enzymology
Female
Humans
In Vitro Techniques
Isoenzymes metabolism
Membrane Proteins
NF-kappa B genetics
Phospholipases A metabolism
Placenta enzymology
Pregnancy
Prostaglandin-Endoperoxide Synthases metabolism
Sulfasalazine pharmacology
Transcription Factor RelA
Dinoprost analogs & derivatives
Extraembryonic Membranes metabolism
NF-kappa B physiology
Phospholipases metabolism
Placenta metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0021-972X
- Volume :
- 89
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- The Journal of clinical endocrinology and metabolism
- Publication Type :
- Academic Journal
- Accession number :
- 15126565
- Full Text :
- https://doi.org/10.1210/jc.2003-031385