title: Kallikrein gene-modified EPCs induce angiogenesis in rats with ischemic hindlimb and correlate with integrin αvβ3 expression.
abstract: BACKGROUND
  Human tissue kallikrein (hTK) plays an essential role in the physiological and pathological mechanisms of blood vessels. This study aimed to determine whether angiogenesis induced by endothelial progenitor cells (EPCs) transduced with the adenovirus-mediated hTK gene could improve blood flow in rat hindlimb ischemia in vivo and to establish a promising mechanism in vitro.
  
  
  METHODS
  EPCs transduced with adenovirus encoding hTK-162 (i.e., Ad/hTK-transduced EPCs or Ad/GFP-transduced EPCs) were administered to Wister rats with hindlimb ischemia through therapeutic neovascularization. Muscular capillary density (MCD), blood flow (BF), and the number of myofibers were measured at days 7, 14, and 21 after treatment. Expressions of integrin αvβ3 and endothelial nitric oxide synthase (eNOS) were detected on the surface of EPCs.
  
  
  RESULTS
  MCD, BF, and the number of myofibers in rats with Ad/hTK-transduced EPCs remarkably increased at day 21 after treatment compared with rats with Ad/GFP-transduced EPCs or the control group (P<0.01). Expressions of integrin αvβ3 and eNOS protein on the surface of EPCs also increased in rats with Ad/hTK-transduced EPCs. The levels of integrin αvβ3 expression were reduced by PI3K and eNOS blockade, and the inhibitor of integrin αvβ3 abrogated the migration and adhesion of hTK-transduced EPCs (P<0.05).
  
  
  CONCLUSIONS
  hTK gene delivery in vivo improves the natural angiogenic response to ischemia. The ability of hTK gene-transduced EPCs can be enhanced in vitro, in which integrin αvβ3 plays a role in the process.
authors.affiliation: Department of Medicine, Qilu Hospital, Shandong University, Jinan, Shandong Province, China.
authors.forename: Shen Shen
authors.initials: SS
authors.lastname: Fu
authors.forename: Fu Ji
authors.initials: FJ
authors.lastname: Li
authors.forename: Yuan Yuan
authors.initials: YY
authors.lastname: Wang
authors.forename: An Bei
authors.initials: AB
authors.lastname: You
authors.forename: Yi Liang
authors.initials: YL
authors.lastname: Qie
authors.forename: Xiao
authors.initials: X
authors.lastname: Meng
authors.forename: Jian Rui
authors.initials: JR
authors.lastname: Li
authors.forename: Bao Chuan
authors.initials: BC
authors.lastname: Li
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Qing
authors.initials: Q
authors.lastname: Da Li
mesh_terms: D000818:Animals; D004727:Endothelium / Q000473:pathology*; D005260:Female; D006614:Hindlimb / Q000098:blood supply*; D039302:Integrin alphaVbeta3 / Q000235:genetics* / Q000378:metabolism; D007511:Ischemia / Q000235:genetics* / Q000378:metabolism; D007610:Kallikreins / Q000235:genetics*; D009389:Neovascularization, Pathologic; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000473:pathology*; D014162:Transfection
chemical_list: D039302:Integrin alphaVbeta3; D052250:Nitric Oxide Synthase Type III; D007610:Kallikreins
references.citation: Cardiovasc Res. 2009 Jan 1;81(1):159-68
references.pmid: 18806276
references.citation: J Biol Chem. 2008 Oct 31;283(44):30363-75
references.pmid: 18765664
references.citation: Trends Cardiovasc Med. 2005 Feb;15(2):57-63
references.pmid: 15885571
references.citation: Circulation. 2002 Feb 12;105(6):732-8
references.pmid: 11839630
references.citation: Circ Res. 2011 Feb 4;108(3):284-93
references.pmid: 21164105
references.citation: Ann Neurol. 2010 Apr;67(4):488-97
references.pmid: 20437584
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3422-7
references.pmid: 10725398
references.citation: Blood. 2004 Oct 1;104(7):2010-9
references.pmid: 15213103
references.citation: Br J Pharmacol. 1998 Jul;124(6):1286-92
references.pmid: 9720802
references.citation: Arterioscler Thromb Vasc Biol. 1999 May;19(5):1156-61
references.pmid: 10323764
references.citation: Nat Rev Drug Discov. 2007 Apr;6(4):273-86
references.pmid: 17396134
references.citation: Stroke. 1999 Sep;30(9):1925-31; discussion 1931-2
references.pmid: 10471446
references.citation: Am J Pathol. 2010 Jan;176(1):504-15
references.pmid: 19948824
references.citation: Microvasc Res. 2010 May;79(3):200-6
references.pmid: 20144623
references.citation: Hypertension. 2000 Jan;35(1 Pt 1):25-31
references.pmid: 10642270
references.citation: Fibrogenesis Tissue Repair. 2012 Mar 12;5:4
references.pmid: 22410175
references.citation: Nat Rev Mol Cell Biol. 2007 Nov;8(11):857-69
references.pmid: 17895899
references.citation: Atherosclerosis. 2007 Apr;191(2):259-64
references.pmid: 16787646
references.citation: Arterioscler Thromb Vasc Biol. 2000 Nov;20(11):2379-85
references.pmid: 11073841
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11951-6
references.pmid: 12195016
references.citation: Arterioscler Thromb Vasc Biol. 2009 May;29(5):657-64
references.pmid: 19164804
references.citation: Hypertension. 2004 Aug;44(2):203-9
references.pmid: 15238569
references.citation: Microsc Res Tech. 2000 Nov 1;51(3):280-301
references.pmid: 11054877
references.citation: Science. 2008 Jan 11;319(5860):195-8
references.pmid: 18187653
references.citation: Circulation. 2004 May 25;109(20):2454-61
references.pmid: 15148275
references.citation: Acta Med Croatica. 2012 Oct;66 Suppl 1:19-24
references.pmid: 23193816
references.citation: Curr Top Dev Biol. 2004;64:207-38
references.pmid: 15563949
references.citation: Diabetes Care. 2009 Jan;32(1):117-9
references.pmid: 18835949
references.citation: Nat Med. 2001 Apr;7(4):430-6
references.pmid: 11283669
references.citation: Circulation. 2004 Jun 8;109(22):2692-7
references.pmid: 15184293
references.citation: Circ Res. 2008 Aug 29;103(5):536-44
references.pmid: 18658052
references.citation: Nature. 2005 Dec 15;438(7070):937-45
references.pmid: 16355211
references.citation: Dev Biol. 2005 Nov 15;287(2):390-402
references.pmid: 16216232
references.citation: Int Wound J. 2013 Oct;10(5):527-33
references.pmid: 22738265
references.citation: J Vasc Surg. 2004 Mar;39(3):655-60
references.pmid: 14981463
references.citation: J Exp Med. 2005 Jun 6;201(11):1825-35
references.pmid: 15928198
references.citation: J Thromb Haemost. 2010 Jan;8(1):185-93
references.pmid: 19874467
references.citation: Gastroenterology. 2007 Jul;133(1):91-107.e1
references.pmid: 17631135
references.citation: Med Clin (Barc). 2008 Nov 15;131(17):665-9
references.pmid: 19087795
references.citation: Angiogenesis. 2012 Sep;15(3):511-9
references.pmid: 22581517