title: The role of shear-induced transforming growth factor-β signaling in the endothelium.
abstract: OBJECTIVE
  Vascular endothelial cells (ECs) are continuously exposed to blood flow that contributes to the maintenance of vessel structure and function; however, the effect of hemodynamic forces on transforming growth factor-β (TGF-β) signaling in the endothelium is poorly described. We examined the potential role of TGF-β signaling in mediating the protective effects of shear stress on ECs.
  
  
  RESULTS
  Human umbilical vein ECs (HUVECs) exposed to shear stress were compared with cells grown under static conditions. Signaling through the TGF-β receptor ALK5 was inhibited with SB525334. Cells were examined for morphological changes and harvested for analysis by real-time polymerase chain reaction, Western blot analysis, apoptosis, proliferation, and immunocytochemistry. Shear stress resulted in ALK5-dependent alignment of HUVECs as well as attenuation of apoptosis and proliferation compared with static controls. Shear stress led to an ALK5-dependent increase in TGF-β3 and Krüppel-like factor 2, phosphorylation of endothelial NO synthase, and NO release. Addition of the NO donor S-nitroso-N-acetylpenicillamine rescued the cells from apoptosis attributable to ALK5 inhibition under shear stress. Knockdown of TGF-β3, but not TGF-β1, disrupted the HUVEC monolayer and prevented the induction of Krüppel-like factor 2 by shear.
  
  
  CONCLUSIONS
  Shear stress of HUVECs induces TGF-β3 signaling and subsequent activation of Krüppel-like factor 2 and NO, and represents a novel role for TGF-β3 in the maintenance of HUVEC homeostasis in a hemodynamic environment.
authors: From the Departments of Ophthalmology (T.E.W., N.G.d.P., P.A.D.) and Pathology (P.A.D.), Schepens Eye Research Institute, Massachusetts Eye and Ear, Harvard Medical School, Boston; and La Jolla Bioengineering Institute, San Diego, CA (N.G.d.P.).
authors: Tony E
authors: TE
authors: Walshe
authors: Nathaniel G
authors: NG
authors: dela Paz
authors: Patricia A
authors: PA
authors: D'Amore
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000503:physiopathology*; D006439:Hemodynamics / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011810:Quinoxalines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013314:Stress, Mechanical; D053782:Transforming Growth Factor beta3 / Q000378:metabolism*
chemical_list: C521813:6-(2-tert-butyl-5-(6-methylpyridin-2-yl)-1H-imidazol-4-yl)quinoxaline; D007093:Imidazoles; C096161:KLF2 protein, human; D051741:Kruppel-Like Transcription Factors; D011810:Quinoxalines; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; C509044:TGFB3 protein, human; D053782:Transforming Growth Factor beta3; D009569:Nitric Oxide; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human
keywords: KLF2 protein, human; endothelium; hemodynamics; nitric oxide; transforming growth factor beta
references: Blood. 2002 Sep 1;100(5):1689-98
references: 12176889
references: J Vasc Surg. 2005 May;41(5):869-80
references: 15886673
references: Angiogenesis. 2001;4(1):11-20
references: 11824373
references: J Biol Chem. 1996 Nov 29;271(48):30656-62
references: 8940041
references: Circ Res. 2005 Mar 18;96(5):e48-57
references: 15718498
references: Circ Res. 2000 Feb 4;86(2):185-90
references: 10666414
references: Circulation. 2003 Dec 2;108(22):2819-25
references: 14638551
references: Circulation. 2007 Nov 13;116(20):2288-97
references: 17967972
references: Int J Mol Med. 2000 Jun;5(6):635-41
references: 10812015
references: J Clin Invest. 1995 Mar;95(3):1363-9
references: 7883983
references: PLoS One. 2009;4(4):e5149
references: 19340291
references: Circ Res. 2008 Aug 1;103(3):289-97
references: 18583708
references: Circ Res. 2007 Jun 22;100(12):1686-95
references: 17585076
references: Arterioscler Thromb Vasc Biol. 2007 Mar;27(3):532-9
references: 17194892
references: Pharmacol Ther. 2010 Dec;128(3):499-508
references: 20826180
references: J Exp Med. 1997 Feb 17;185(4):601-7
references: 9034139
references: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9385-9
references: 10920209
references: Nat Rev Mol Cell Biol. 2007 Nov;8(11):857-69
references: 17895899
references: Birth Defects Res A Clin Mol Teratol. 2005 Dec;73(12):956-65
references: 16323168
references: Arterioscler Thromb Vasc Biol. 2001 May;21(5):785-90
references: 11348875
references: Circ Res. 2003 Aug 22;93(4):354-63
references: 12893742
references: Circ Res. 1998 Feb 9;82(2):186-94
references: 9468189
references: Arterioscler Thromb Vasc Biol. 2009 Nov;29(11):1902-8
references: 19696400
references: J Biol Chem. 1999 Feb 12;274(7):4213-9
references: 9933619
references: Nature. 1997 Mar 27;386(6623):338
references: 9121550
references: Neuroscience. 2011 Aug 25;189:123-31
references: 21664439
references: Invest Ophthalmol Vis Sci. 2005 Jan;46(1):375-82
references: 15623798
references: Circ Res. 2005 Apr 1;96(6):684-92
references: 15718503
references: J Lab Clin Med. 2004 Sep;144(3):148-55
references: 15454884
references: Br J Pharmacol. 2002 Dec;137(7):1021-30
references: 12429574
references: Arterioscler Thromb Vasc Biol. 2009 Aug;29(8):1185-92
references: 19461051
references: J Clin Invest. 2006 Jan;116(1):49-58
references: 16341264
references: Thorax. 2001 Jul;56(7):549-56
references: 11413354
references: Nat Med. 2006 Jun;12(6):642-9
references: 16751767
references: Cell Tissue Res. 1999 Oct;298(1):85-93
references: 10555542
references: Cancer Cell. 2007 Oct;12(4):293-5
references: 17936552
references: Eur J Vasc Endovasc Surg. 1995 Jan;9(1):86-92
references: 7664019
references: Am J Pathol. 2005 Aug;167(2):609-18
references: 16049344
references: Hypertension. 1999 Jun;33(6):1359-63
references: 10373216
references: Methods Mol Biol. 2011;769:137-47
references: 21748674
references: Blood. 2010 Mar 25;115(12):2533-42
references: 20032497
references: Cardiovasc Res. 2005 Oct 1;68(1):155-64
references: 15907823
references: J Biol Chem. 1997 Jul 18;272(29):18000-6
references: 9218427
references: Surv Ophthalmol. 1994 Sep-Oct;39(2):123-32
references: 7801220
references: J Biol Chem. 2010 May 7;285(19):14806-14
references: 20207738
references: Am J Physiol Heart Circ Physiol. 2003 Aug;285(2):H631-6
references: 12750062
references: J Exp Med. 2008 Feb 18;205(2):491-501
references: 18268040
references: J Cell Sci. 2007 May 15;120(Pt 10):1810-7
references: 17502485
references: Diabetes. 2007 Nov;56(11):2690-6
references: 17682093
references: Microvasc Res. 2010 Jul;80(1):166-73
references: 20226797
references: Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13176-81
references: 8917564
references: J Biol Chem. 2001 Aug 10;276(32):29632-43
references: 11382746
references: Blood. 2006 Jun 1;107(11):4354-63
references: 16455954
references: Cytokine Growth Factor Rev. 2011 Jun;22(3):167-75
references: 21700485
references: Blood. 2007 May 15;109(10):4249-57
references: 17244683
references: Proc Natl Acad Sci U S A. 2004 Feb 3;101(5):1386-91
references: 14742866
references: J Exp Med. 2004 May 17;199(10):1305-15
references: 15136591
references: J Cell Biol. 1998 May 4;141(3):805-14
references: 9566978
references: Dev Dyn. 2011 Jul;240(7):1670-80
references: 21604321
references: Pflugers Arch. 2010 Nov;460(6):965-74
references: 20697735
references: J Vasc Res. 2004 Nov-Dec;41(6):491-8
references: 15528931
references: Arterioscler Thromb Vasc Biol. 2009 Sep;29(9):1251-7
references: 19325140
references: Arterioscler Thromb Vasc Biol. 2005 Jun;25(6):1135-41
references: 15817882
references: J Biol Chem. 2009 Mar 27;284(13):8412-20
references: 19047056
references: Cardiovasc Res. 2011 Feb 15;89(3):661-70
references: 21030535
references: Nature. 1995 Sep 21;377(6546):239-42
references: 7545787