title: N-Acetyl-cysteine inhibits phospholipid metabolism, proinflammatory cytokine release, protease activity, and nuclear factor-kappaB deoxyribonucleic acid-binding activity in human fetal membranes in vitro.
abstract: The production of reactive oxygen species (ROS), prostaglandins (PGs), proinflammatory cytokines, and proteases has been implicated in the pathogenesis of term and preterm labor. The nuclear factor-kappaB (NF-kappaB) transcription pathway is activated by ROS and is a key regulator of PGs, proinflammatory cytokine release, and protease activity. N-Acetyl-cysteine (NAC) is an antioxidant that through its ability to scavenger ROS suppresses NF-kappaB DNA-binding activity and resultant gene expression. The aim of this study was to elucidate the effect of NAC on NF-kappaB DNA-binding activity, phospholipid metabolism, cytokine release, and protease activity from human fetal membranes. Human amnion and choriodecidua (n = 9 separate placentas) were treated with 0 (control), 5, 10, or 15 mM NAC in the presence of 10 micro g/ml lipopolysaccharide. After 6-h incubation, the tissues were collected, NF-kappaB DNA binding activity was assessed by gel shift binding assays, and matrix metalloproteinase-9 and urokinase-type plasminogen activator activity were determined by zymography. The incubation medium was collected and assayed for type II phospholipase A(2) tissue content, IL-6, IL-8, TNFalpha, and 8-isoprostane release by ELISA. The release of PGF(2alpha) was measured by RIA. Treatment of fetal membranes with NAC significantly suppressed lipopolysaccharide-stimulated type II phospholipase A(2) release and content; PGF(2alpha), IL-6, IL-8, TNFalpha, and 8-isoprostane release; and matrix metalloproteinase-9 and urokinase-type plasminogen activator enzyme activity and suppressed NF-kappaB DNA-binding activity (by ANOVA, P < 0.05). The data presented in this study demonstrate that NAC inhibits an NF-kappaB-activated pathway and subsequent phospholipid metabolism, proinflammatory cytokine release, and protease activity in human fetal membranes.
authors.affiliation: Department of Obstetrics and Gynecology, University of Melbourne, and Mercy Perinatal Research Center, Mercy Hospital for Women, East Melbourne, 3002 Victoria, Australia. mlappas@unimelb.edu.au
authors.forename: Martha
authors.initials: M
authors.lastname: Lappas
authors.forename: Michael
authors.initials: M
authors.lastname: Permezel
authors.forename: Gregory E
authors.initials: GE
authors.lastname: Rice
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000650:Amnion / Q000187:drug effects / Q000378:metabolism; D002823:Chorion / Q000187:drug effects / Q000378:metabolism; D046508:Culture Techniques; D016207:Cytokines / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D003656:Decidua / Q000187:drug effects / Q000378:metabolism; D015237:Dinoprost / Q000378:metabolism / Q000031:analogs & derivatives; D010450:Endopeptidases / Q000378:metabolism*; D005321:Extraembryonic Membranes / Q000187:drug effects* / Q000378:metabolism; D028441:F2-Isoprostanes / Q000378:metabolism; D005260:Female; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018384:Oxidative Stress; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D011247:Pregnancy; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D016207:Cytokines; D028441:F2-Isoprostanes; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D010743:Phospholipids; C075750:8-epi-prostaglandin F2alpha; D004247:DNA; D015237:Dinoprost; D007770:L-Lactate Dehydrogenase; D010741:Phospholipases A; D010450:Endopeptidases; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9; D000111:Acetylcysteine