title: Arachidonic acid regulation of the cytosolic phospholipase A 2α/cyclooxygenase-2 pathway in mouse endometrial stromal cells.
abstract: OBJECTIVE
  To investigate the role of arachidonic acid (AA) in mouse endometrial stromal cells.
  
  
  METHODS
  Experimental animal study.
  
  
  METHODS
  University research laboratory.
  
  
  METHODS
  Sexually mature female CD1-strain mice.
  
  
  METHODS
  Primary culture of endometrial stromal cells.
  
  
  METHODS
  Western blot and real-time polymerase chain reaction for gene expression and/or phosphorylation analysis. Luciferase assay for Cox-2 promoter analysis.
  
  
  RESULTS
  AA-derived prostaglandins play important roles during embryo implantation and decidualization. However, the function of AA itself in reproduction is largely unknown. In this study, exogenous AA stimulated cPLA(2α) phosphorylation and COX-2 expression, mainly through ERK1/2 in mouse endometrial stromal cells, and p38 inhibitor modestly inhibited cPLA(2α) phosphorylation induced by AA. The induction of COX-2 by AA was diminished by short interfering RNA against C/EBPβ and inhibitory C/EBPβ (LIP). C/EBPβ binding site at -872--864 of Cox-2 promoter contributes to Cox-2 promoter activation induced by C/EBPβ transfection. The expression of C/EBPβ protein induced by AA was inhibited by p38 inhibitor, and the phosphorylation of C/EBPβ induced by AA was inhibited by p38 inhibitor and ERK1/2 inhibitor. A nonmetabolized analogue of AA (ETYA) also enhanced cPLA(2α) phosphorylation and COX-2 expression. The activation of cPLA(2α)/COX-2 by AA was not inhibited by COX inhibitor indomethacin.
  
  
  CONCLUSIONS
  AA can induce cPLA(2α)/COX-2 pathway activation in mouse endometrial stromal cells.
authors.affiliation: Department of Life Science, Shantou University, Shantou, People's Republic of China.
authors.forename: Zhen-Ao
authors.initials: ZA
authors.lastname: Zhao
authors.forename: Zhi-Rong
authors.initials: ZR
authors.lastname: Zhang
authors.forename: Xiu
authors.initials: X
authors.lastname: Xu
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Deng
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.forename: Jing-Yu
authors.initials: JY
authors.lastname: Leng
authors.forename: Xiao-Huan
authors.initials: XH
authors.lastname: Liang
authors.forename: Zeng-Ming
authors.initials: ZM
authors.lastname: Yang
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D015153:Blotting, Western; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000096:biosynthesis* / Q000235:genetics; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology; D004717:Endometrium / Q000166:cytology / Q000201:enzymology*; D004790:Enzyme Induction; D005260:Female; D054513:Group IV Phospholipases A2 / Q000378:metabolism*; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012741:Sexual Maturation; D015398:Signal Transduction; D017154:Stromal Cells / Q000201:enzymology*; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; C545013:Cebpb protein, mouse; D052246:Cyclooxygenase 2 Inhibitors; C516390:Pla2g4a protein, mouse; D047428:Protein Kinase Inhibitors; D016718:Arachidonic Acid; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; C535148:Mapk1 protein, mouse; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D054513:Group IV Phospholipases A2