title: Thrombin-induced chemokine production in endometrial stromal cells.
abstract: BACKGROUND
  In order to investigate the regulation of chemokines [interleukin-8 (IL-8), growth-regulated oncogene (GRO)α, monocyte chemoattractant protein-1 (MCP-1)) induced by thrombin in endometrial stromal cells (ESCs), the effects of thrombin, a protease activated receptor (PAR)-1 antagonist (PPACK), mitogen-activated protein kinase kinase inhibitor (U0126), phospholipase C inhibitor (U-73122), an antagonist of the intracellular InsP3 receptor (2-aminoethoxy-diphenylborate (2-APB)] and a protein kinase C inhibitor (GF-109203X) on the production of chemokines by ESCs were evaluated.
  
  
  METHODS
  ESCs from eight endometrial specimens in the secretory phase were cultured and incubated for 24h with thrombin and PPACK, U0126, U-73122, 2-APB or GF-109203X. The levels of IL-8, GROα and MCP-1 in the culture medium were measured by means of ELISA. The activation of MAP kinase was detected by western blot analysis using anti-phosphorylated MAP kinase (ERK1/2) antibody.
  
  
  RESULTS
  Following stimulation by thrombin, the production of IL-8, GROα and MCP-1 increased significantly in a dose-dependent manner. PPACK, U0126, U-73122, 2-APB or GF-109203X suppressed the increases in production of IL-8, GROα and MCP-1 induced by thrombin (P < 0.001, P <0.001 and P <0.001, respectively). MAP kinase activities were induced by treatment with thrombin, and were suppressed by PPACK, U0126, U-73122, 2-APB or GF-109203X.
  
  
  CONCLUSIONS
  Our results suggest that thrombin stimulates the production of IL-8, GROα and MCP-1 via PAR-1 by a mechanism involving the MAP kinase system. The increases in IL-8, GROα and MCP-1 may contribute to the maintenance of implantation involving leukocyte chemotaxis.
authors.affiliation: Department of Obstetrics and Gynecology, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama, Yufu, Oita 879-5593, Japan. kawayas@oita-u.ac.jp
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kawano
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Furukawa
authors.forename: Yukie
authors.initials: Y
authors.lastname: Kawano
authors.forename: Kaei
authors.initials: K
authors.lastname: Nasu
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narahara
mesh_terms: D000328:Adult; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D001896:Boron Compounds / Q000494:pharmacology; D002070:Butadienes / Q000494:pharmacology; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000096:biosynthesis*; D054360:Chemokine CXCL1 / Q000096:biosynthesis; D004717:Endometrium / Q000166:cytology; D004963:Estrenes / Q000494:pharmacology; D005260:Female; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D016209:Interleukin-8 / Q000096:biosynthesis*; D008301:Maleimides / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology*; D010975:Platelet Aggregation Inhibitors
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D001896:Boron Compounds; D002070:Butadienes; C475829:CXCL1 protein, human; D018932:Chemokine CCL2; D054360:Chemokine CXCL1; D004963:Estrenes; D007211:Indoles; D016209:Interleukin-8; D008301:Maleimides; D009570:Nitriles; D011760:Pyrrolidinones; C113580:U 0126; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; C109986:2-aminoethoxydiphenyl borate; D013917:Thrombin; C070515:bisindolylmaleimide I; C040308:phenylalanyl-prolyl-arginine-chloromethyl ketone; D010975:Platelet Aggregation Inhibitors