title: G protein-coupled estrogen receptor1 (GPER1) may mediate Rho-kinase (ROCK-2) up-regulation in coronary endothelial cells.
abstract: OBJECTIVE
  Effect of estrogenic compounds and 17β-estradiol (E2), which induces endothelial cell motility, was investigated on ROCK-2 expression in rat coronary vascular endothelial cells (CVEC).
  
  
  METHODS
  The CVEC were isolated from the heart of Wistar rats by collagenase (0.04%) and incubated with E2 (1-100 nM), estrogen receptor α (ERα) agonist: propyl pyrazole triol (PPT, 10 nM); ERβ agonists: (2,3-bis(4-hydroxyphenyl)-propionitrile, DPN, 10 nM) and E2-conjugate with bovine serum albumin (E2-BSA, 1 nM); and GPER1 agonist: G1 (100 nM). Furthermore, the effect of combination of E2 with estrogen receptors (ERs) antagonist and GPER1 agonist, ICI-182780 (10 µM), physiological estrogen antagonists: progesterone (P4, 10-100 nM) and testosterone (T, 10-100 nM); transcription inhibitor: actinomycin-D (1 µg/ml); GPER1 antagonist: G-15 (100 nM), superoxide dismutase, (SOD, 500 U/ml); Gi/o protein inhibitor: pertussis toxin (PTX, 100 µg/ml); and epidermal growth factor receptor (EGFR) blocker: AG-1478 (10 µM) was tested. After 24h incubation, ROCK-2 and GPER1 protein expressions were detected in the CVEC by Western-blotting.
  
  
  RESULTS
  E2, ICI-182780, and G1 but not E2-BSA significantly up-regulated ROCK-2 expression, which was suppressed by actinomycin-D, PTX, AG-1478, and G-15. However, PPT and DPN had no effects on the ROCK-2 expression. ICI-182780, P4, T or SOD did not antagonize the E2 action. GPER1 expression was demonstrated in the CVEC.
  
  
  CONCLUSIONS
  Estrogens could up-regulate ROCK-2 in the rat CVEC through GPER1 and EGFR transactivation.
authors: Department of Pharmacology, Mersin University, Mersin, Turkey.
authors: A H
authors: AH
authors: Kurt
authors: R N
authors: RN
authors: Tiftik
authors: I
authors: I
authors: Un
authors: S
authors: S
authors: Ulker
authors: K
authors: K
authors: Buyukafsar
mesh_terms: D000818:Animals; D052117:Benzodioxoles / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003326:Coronary Circulation / Q000502:physiology*; D003609:Dactinomycin / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D000077267:Fulvestrant; D008297:Male; D009569:Nitric Oxide / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D010636:Phenols; D011374:Progesterone / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011720:Pyrazoles / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013739:Testosterone / Q000494:pharmacology; D020032:Tyrphostins / Q000494:pharmacology; D015854:Up-Regulation / Q000502:physiology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011422:Propionates
chemical_list: D052117:Benzodioxoles; D003609:Dactinomycin; D004791:Enzyme Inhibitors; D066246:ErbB Receptors; D004958:Estradiol; D004965:Estrogen Antagonists; D000077267:Fulvestrant; D009569:Nitric Oxide; D009570:Nitriles; D010636:Phenols; D011374:Progesterone; D011500:Protein Synthesis Inhibitors; D011720:Pyrazoles; D011799:Quinazolines; D011804:Quinolines; D043562:Receptors, G-Protein-Coupled; D013739:Testosterone; D020032:Tyrphostins; D054460:rho-Associated Kinases; C486182:2,3-bis(4-hydroxyphenyl)-propionitrile; D011422:Propionates; C568862:4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta(c)quinoline; C520305:Gper1 protein, rat; C486184:4,4',4''-(4-propyl-((1)H)-pyrazole-1,3,5-triyl) tris-phenol; C101044:RTKI cpd; C580818:ROCK2 protein, rat