Gene delivery of soluble vascular endothelial growth factor receptor-1 (sFlt-1) inhibits intra-plaque angiogenesis and suppresses development of atherosclerotic plaque.
Intra-plaque angiogenesis plays an important role in the development of atherosclerotic plaque. Vascular endothelial growth factor (VEGF) is a major initiating factor in this pathologic progress. One selective and specific inhibitor of VEGF is soluble VEGF receptor-1 (sFlt-1). The anti-angiogenic utilization of sFlt-1 in treatment of atherosclerotic plaque has not been fully confirmed yet. Our study was designed to construct eukaryotic expression recombinant pEGFP-N1-sFlt-1, evaluate sFlt-1 recombinant's effects on endothelial cells proliferation and tube formation in vitro, and investigate effects of local high-expressed sFlt-1 on atherosclerotic plaque in vivo. Rabbit models of atherosclerotic plaque were established by high-lipid diet combined with injury induced by balloon catheter on iliac artery intima. Animals were divided into four groups randomly: control group (C), atherosclerotic plaque group (AP), atherosclerotic plaque with blank vector pEGFP-N1 transfection group (APV), and atherosclerotic plaque with pEGFP-N1-sFlt-1 transfection group (APsFlt-1). The local expression of sFlt-1 protein in target artery was detected by western blotting. The plaque area (PA), plaque circumference (PC), and maximum plaque thickness (MPT) were measured via HE staining. Degree of intra-plaque angiogenesis was evaluated by CD34+ cells immunohistochemistry. As results, we observed that pEGFP-N1-sFlt-1 transfection suppressed the HUVECs proliferation and ability of tube formation, against the effect of VEGF. We obtained higher local expression of sFlt-1 protein in Group APsFlt-1 than that in other groups (P < 0.05). PA, PC, and MPT of plaque in group APsFlt-1 were significantly decreased when compared with other groups (P < 0.05). Amount of annulations surrounded by CD34-positive cells was significantly decreased in pEGFP-N1-sFlt-1 transfection group, which represented decreased level of intra-plaque neovessels formation. The present study confirmed that local gene delivery of sFlt-1 can suppress plaque formation, as one of possible mechanisms, via inhibitive effect on intra atherosclerotic plaque angiogenesis, which hints at the clinical utility of sFlt-1 in atherosclerosis therapy.
Wang
Yaosheng
Y
Department of Cardiovascular Disease, Second Affiliated Hospital, Nanchang University, Jiangxi Province, People's Republic of China.
Zhou
Yihua
Y
He
Lipeng
L
Hong
Kui
K
Su
Hai
H
Wu
Yanqing
Y
Wu
Qinghua
Q
Han
Mihan
M
Cheng
Xiaoshu
X
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000502:physiology; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D058226:Plaque, Atherosclerotic / Q000473:pathology* / Q000628:therapy*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics* / Q000378:metabolism*
D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D040281:Vascular Endothelial Growth Factor Receptor-1
Arterioscler Thromb Vasc Biol. 2005 Oct;25(10):2054-61
16037567
Nature. 2002 Jun 13;417(6890):750-4
12066187
Proc Natl Acad Sci U S A. 2006 Oct 17;103(42):15491-6
17030814
J Am Coll Cardiol. 2009 Apr 28;53(17):1517-27
19389562
J Gene Med. 2009 May;11(5):422-34
19266483
Am J Physiol Heart Circ Physiol. 2008 Oct;295(4):H1377-84
18660448
Nature. 2006 Oct 26;443(7114):993-7
17051153
Circulation. 2002 Mar 5;105(9):1110-5
11877364
J Pathol. 2009 May;218(1):7-29
19309025
Circulation. 2004 Nov 2;110(18):2843-50
15505090
Immunity. 2004 Dec;21(6):831-42
15589171
Clin Sci (Lond). 2005 Sep;109(3):227-41
16104843
J Biol Chem. 2008 Dec 5;283(49):34250-9
18922791
Biol Reprod. 2002 Jun;66(6):1768-74
12021060
Oncol Rep. 2005 Dec;14(6):1565-9
16273257
J Am Coll Cardiol. 2007 May 29;49(21):2073-80
17531655
JACC Cardiovasc Imaging. 2009 May;2(5):616-24
19442950
Ann Med. 2009;41(4):257-64
19089693
Heart Vessels. 2009 Sep;24(5):321-8
19784813
J Vasc Surg. 2007 Jan;45(1):155-9
17210401
Cold Spring Harb Symp Quant Biol. 2002;67:471-82
12858573
Arterioscler Thromb Vasc Biol. 2007 May;27(5):1095-100
17303776
Am J Physiol Heart Circ Physiol. 2004 Nov;287(5):H2346-51
15178545
J Am Coll Cardiol. 2001 Jun 15;37(8):2126-30
11419898
Stroke. 2006 Jun;37(6):1572-7
16627791
Curr Drug Targets Inflamm Allergy. 2005 Feb;4(1):3-8
15720228
Circulation. 1998 Nov 17;98(20):2108-16
9815864
Heart Vessels. 2009 Jul;24(4):254-9
19626396
J Clin Pathol. 2004 Mar;57(3):266-72
14990597