title: RhoA-Rho kinase and platelet-activating factor stimulation of ovine foetal pulmonary vascular smooth muscle cell proliferation.
abstract: OBJECTIVE
  Platelet-activating factor (PAF) is produced by pulmonary vascular smooth muscle cells (PVSMC). We studied effects of Rho kinase on PAF stimulation of PVSMC proliferation in an attempt to understand the role of RhoA/Rho kinase on PAF-induced ovine foetal pulmonary vascular remodelling. Our hypothesis is that PAF acts through Rho kinase, as one of its downstream signals, to induce arterial (SMC-PA) and venous (SMC-PV) cell proliferation in the hypoxic lung environment of the foetus, in utero.
  
  
  METHODS
  Rho kinase and MAPK effects on PAF receptor (PAFR)-mediated cell population expansion, and PAFR expression, were studied by DNA synthesis, western blot analysis and immunocytochemistry. Effects of constructs T19N and G14V on PAF-induced cell proliferation were also investigated.
  
  
  RESULTS
  Hypoxia increased PVSMC proliferation and Rho kinase inhibitors, Y-27632 and Fasudil (HA-1077) as well as MAPK inhibitors PD 98059 and SB 203580 attenuated PAF stimulation of cell proliferation. RhoA T19N and G14V stimulated cell proliferation, but co-incubation with PAF did not affect proliferative effects of the constructs. PAFR protein expression was significantly downregulated in both cell types by both Y-27632 and HA-1077, with comparable profiles. Also, cells treated with Y-27632 had less PAF receptor fluorescence with significant disruption of cell morphology.
  
  
  CONCLUSIONS
  Our results show that Rho kinase non-specifically modulated PAFR-mediated responses by a translational modification of PAFR protein, and suggest that, in vivo, activation of Rho kinase by PAF may be a further pathway to sustain PAFR-mediated PVSMC proliferation.
authors.affiliation: Division of Neonatology, Department of Pediatrics, Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, Torrance, CA, 90502, USA.
authors.forename: L S
authors.initials: LS
authors.lastname: Renteria
authors.forename: M
authors.initials: M
authors.lastname: Austin
authors.forename: M
authors.initials: M
authors.lastname: Lazaro
authors.forename: M A
authors.initials: MA
authors.lastname: Andrews
authors.forename: J
authors.initials: J
authors.lastname: Lustina
authors.forename: J U
authors.initials: JU
authors.lastname: Raj
authors.forename: B O
authors.initials: BO
authors.lastname: Ibe
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000577:Amides / Q000494:pharmacology; D000818:Animals; D015687:Cell Hypoxia; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D005419:Flavonoids / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D008168:Lung / Q000166:cytology*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D009154:Mutation; D010972:Platelet Activating Factor / Q000494:pharmacology*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D012756:Sheep; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000577:Amides; D005419:Flavonoids; D007093:Imidazoles; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D011725:Pyridines; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor; C108830:Y 27632; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; D020742:rhoA GTP-Binding Protein; C093642:SB 203580; C049347:fasudil; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
references.citation: Crit Rev Oral Biol Med. 2000;11(2):240-58
references.pmid: 12002818
references.citation: Rev Physiol Biochem Pharmacol. 1999;134:201-34
references.pmid: 10087910
references.citation: Crit Care Med. 2007 Jul;35(7):1756-62
references.pmid: 17522574
references.citation: J Biol Chem. 1998 Feb 6;273(6):3285-90
references.pmid: 9452444
references.citation: Genes Dev. 1998 Jan 15;12(2):149-62
references.pmid: 9436976
references.citation: Int Angiol. 1996 Jun;15(2):100-3
references.pmid: 8803632
references.citation: Am J Physiol Gastrointest Liver Physiol. 2006 Jan;290(1):G83-95
references.pmid: 16179599
references.citation: J Clin Invest. 1999 Mar;103(5):691-6
references.pmid: 10074486
references.citation: Curr Opin Pharmacol. 2009 Jun;9(3):287-96
references.pmid: 19297247
references.citation: J Mol Med (Berl). 2002 Oct;80(10):629-38
references.pmid: 12395147
references.citation: Am J Physiol Lung Cell Mol Physiol. 2010 Dec;299(6):L872-8
references.pmid: 20709731
references.citation: Annu Rev Physiol. 1995;57:115-34
references.pmid: 7778860
references.citation: Am J Physiol Lung Cell Mol Physiol. 2008 Nov;295(5):L828-36
references.pmid: 18689606
references.citation: EMBO J. 2003 Mar 17;22(6):1313-24
references.pmid: 12628924
references.citation: Physiol Rev. 2010 Oct;90(4):1291-335
references.pmid: 20959617
references.citation: Am J Physiol Lung Cell Mol Physiol. 2005 May;288(5):L879-86
references.pmid: 15618453
references.citation: Cell Prolif. 2008 Apr;41(2):208-29
references.pmid: 18336468
references.citation: Am J Physiol Heart Circ Physiol. 2007 Jun;292(6):H2773-81
references.pmid: 17322418
references.citation: Am J Physiol Lung Cell Mol Physiol. 2006 Nov;291(5):L976-82
references.pmid: 16815887
references.citation: J Cell Commun Signal. 2012 Dec;6(4):205-16
references.pmid: 22878922
references.citation: Physiol Rev. 2000 Oct;80(4):1669-99
references.pmid: 11015622
references.citation: J Appl Physiol (1985). 1992 Sep;73(3):1084-92
references.pmid: 1400021
references.citation: Am J Physiol Lung Cell Mol Physiol. 2005 Mar;288(3):L480-7
references.pmid: 15531748
references.citation: Biochim Biophys Acta. 1994 Nov 10;1224(2):323-8
references.pmid: 7981248
references.citation: J Appl Physiol (1985). 1992 Dec;73(6):2559-66
references.pmid: 1490969
references.citation: Circ Res. 2006 Feb 17;98(3):322-34
references.pmid: 16484628
references.citation: Nature. 2002 Dec 12;420(6916):629-35
references.pmid: 12478284
references.citation: Physiol Rev. 2001 Jul;81(3):999-1030
references.pmid: 11427690
references.citation: Am J Physiol Heart Circ Physiol. 2006 Jun;290(6):H2528-34
references.pmid: 16399861
references.citation: J Biol Chem. 1996 Sep 20;271(38):23146-53
references.pmid: 8798508
references.citation: Cell Prolif. 2007 Dec;40(6):949-60
references.pmid: 18021181
references.citation: Am J Physiol Lung Cell Mol Physiol. 2004 Oct;287(4):L656-64
references.pmid: 14977625
references.citation: Br J Pharmacol. 1995 Jan;114(1):203-9
references.pmid: 7712019
references.citation: Am J Physiol. 1995 Aug;269(2 Pt 2):H704-9
references.pmid: 7653635
references.citation: Oncol Lett. 2013 Apr;5(4):1417-1421
references.pmid: 23599805
references.citation: Am J Physiol Lung Cell Mol Physiol. 2004 Jul;287(1):L250-7
references.pmid: 15064228
references.citation: J Biol Chem. 1996 Sep 20;271(38):23298-303
references.pmid: 8798529
references.citation: Am J Respir Crit Care Med. 2005 Mar 1;171(5):494-9
references.pmid: 15563635
references.citation: Respir Physiol. 1992 Apr-May;88(1-2):129-40
references.pmid: 1626132
references.citation: J Vasc Surg. 2012 Aug;56(2):446-54
references.pmid: 22521802
references.citation: Am J Physiol Lung Cell Mol Physiol. 2007 Dec;293(6):L1475-82
references.pmid: 17951313
references.citation: J Cell Biol. 2002 Mar 4;156(5):893-903
references.pmid: 11877460
references.citation: Eur Respir J. 2009 Nov;34(5):1100-10
references.pmid: 19324947
references.citation: Am Rev Respir Dis. 1990 Dec;142(6 Pt 1):1258-62
references.pmid: 2252241
references.citation: Exp Lung Res. 2009 May;35(4):324-43
references.pmid: 19415549
references.citation: Mol Genet Metab. 2010 Dec;101(4):400-8
references.pmid: 20813571
references.citation: J Appl Physiol (1985). 1998 Sep;85(3):1079-85
references.pmid: 9729586
references.citation: Mol Genet Metab. 2002 Nov;77(3):237-48
references.pmid: 12409272