Nitric oxide is an upstream signal of vascular endothelial growth factor-induced extracellular signal-regulated kinase1/2 activation in postcapillary endothelium

被引:385
作者
Parenti, A
Morbidelli, L
Cui, XL
Douglas, JG
Hood, JD
Granger, HJ
Ledda, F
Ziche, M
机构
[1] Univ Florence, Dept Preclin & Clin Pharmacol, I-50134 Florence, Italy
[2] Case Western Reserve Univ, Sch Med, Div Hypertens, Cleveland, OH 44106 USA
[3] Texas A&M Univ, Hlth Sci Ctr, Microcirculat Res Inst, College Stn, TX 77843 USA
[4] Texas A&M Univ, Hlth Sci Ctr, Dept Med Physiol, College Stn, TX 77843 USA
关键词
D O I
10.1074/jbc.273.7.4220
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We recently demonstrated that nitric oxide (NO) significantly contributes to the mitogenic effect of vascular endothelial growth factor (VEGF), suggesting a role for the NO pathway in the signaling cascade following kinase-derivative receptor activation in vascular endothelium. The aim of this study was to investigate the intracellular pathways linked to VEGF/NO-induced endothelial cell proliferation, We assessed the activity of the mitogen-activated protein kinase (MAPK) that is specifically activated by growth factors, extracellular-regulated kinase (ERK1/2), on cultured microvascular endothelium isolated from coronary postcapillary venules, ERK1/2 was immunoprecipitated, and its activity was assessed with an immunocomplex kinase assay. in endothelial cells exposed for 5 min to the NO donor drug sodium nitroprusside at a concentration of 100 mu M, ERK1/2 activity significantly increased. VEGF produced a time-and concentration-dependent activation of ERK1/2. Maximal activity was obtained after 5 min of stimulation at a concentration of 10 ng/ml., The specific MAPK kinase inhibitor PD 98059 abolished ERK1/2 activation and endothelial cell proliferation in a concentration-dependent manner in response to VEGF and sodium nitroprusside. The NO synthase inhibitor N-omega-monomethyl-L-arginine, as well as the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one blocked the activation of ERK1/2 induced by VEGF, suggesting that NO and cGMP contributed to the VEGF-dependent ERK1/2 activation, These results demonstrate for the first time that kinase-derivative receptor activation triggers the NO synthase/guanylate cyclase pathway to activate the MAPK cascade and substantiates the hypothesis that the activation of ERK1/2 is necessary for VEGF-induced endothelial cell proliferation.
引用
收藏
页码:4220 / 4226
页数:7
相关论文
共 45 条
[1]  
ADLER H, 1995, J IMMUNOL, V154, P4710
[2]   SELECTIVE REQUIREMENT FOR MAP KINASE ACTIVATION IN THYMOCYTE DIFFERENTIATION [J].
ALBEROLAILA, J ;
FORBUSH, KA ;
SEGER, R ;
KREBS, EG ;
PERLMUTTER, RM .
NATURE, 1995, 373 (6515) :620-623
[3]   PD-098059 IS A SPECIFIC INHIBITOR OF THE ACTIVATION OF MITOGEN-ACTIVATED PROTEIN-KINASE KINASE IN-VITRO AND IN-VIVO [J].
ALESSI, DR ;
CUENDA, A ;
COHEN, P ;
DUDLEY, DT ;
SALTIEL, AR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (46) :27489-27494
[4]   Vascular endothelial growth factor increases hydraulic conductivity of isolated perfused microvessels [J].
Bates, DO ;
Curry, FE .
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY, 1996, 271 (06) :H2520-H2528
[5]   SIGNAL-TRANSDUCTION VIA THE MAP KINASES - PROCEED AT YOUR OWN RSK [J].
BLENIS, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (13) :5889-5892
[6]  
BREIER G, 1992, DEVELOPMENT, V114, P521
[7]  
BROCK TA, 1991, AM J PATHOL, V138, P213
[8]   SHEAR-STRESS INDUCED RELEASE OF NITRIC-OXIDE FROM ENDOTHELIAL-CELLS GROWN ON BEADS [J].
BUGA, GM ;
GOLD, ME ;
FUKUTO, JM ;
IGNARRO, LJ .
HYPERTENSION, 1991, 17 (02) :187-193
[9]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2