Lipophilic statin-induced antiangiogenesis requires the inhibition of ras farnesylation in human coronary artery endothelial cells

294A

ABSTRACTS

- Vascular Disease, Hypertension,

and Prevention

JACC

March 19.2003

stratified subjects based on their CIMT Into those with CIMT > 0.6 mm and those with CIMT ( 0.6”“. Differences in the varws markers between the 2 groups are shown

Infusion (36i13 to 52*22% ng/ml vs. 3.25*0.96 to 3.01+0.91% (p=NS for pre vs. post for both). Conclusions ADR administration results in rapid depletion of *NO levels as ewdenced by attenuation of agonist dependent responses in rabbits and reactive hyperemic

below.

responses

There

was also a strong

correlation

0.01). and an inverse correlation

Variable

(Units)

between

between

Eh GSH/GSSG

and CIMT

(I = 0.60, p- <

GSH and CIMT (I = - 0.52, p- < 0.01). We then

CIMT > 0.60 mm

P-value

7.68% (+/-2.4)

5.59% (+/- 3.1)

0.02

CIMT so.60

mm

wti FMD (%) Eh GSHiGSSG

(mV)

GSH (uM) GSSG (uM) After multivariate

analysis,

CIMT. Conclusion:

-126.2 (+/- 6.3)

-118.8 (+/- 7.2)

0.01

2.51(+/-0.86)

1.84 (+i- 0.69)

0.03

0.24 (+/-0.11)

0.23 (+/- 0.12)

both FMD and Eh GSHiGSSG

These flndings

suggest

of oxidative

1177-126

predictors

of

stress such as Eh

GSH/GSSG may be helpful in predicting individuals at risk for early atherosclerosis. predictive value appears to be independent of endothelial function.

POSTER

Fukuoka

University,

inhibitor

Combined

Fibrinolysis Coronary

With

Statin

Background:

Because

tensln converting

Artery

these therapies in hypercholesterolemic Methods: We administered simvastatin 2 months

with washout

Inflammation

2 months

Slmvastatin

effects

and from

P=O.O37, respectively), radical from 1.41iO.66

97+47

with ramipril

plasma

to 79*44

and plasma to 1 .l lr0.60

of statins

differ, we studied the vascular

to 30 hypercholesterolemic

(both P
uM by 4*58%

patients

and antio-

responses

to

with CAD. This

crcsscver

significantly

were

seeded

on a matrix

and without

mews,

cells were cultured

endothelial

Lipophlllc

transduction.

promotion

of HCECs

gel and grown

cell growth

in the presence

statins (slmvastatln,

Therefore,

of angiogenesis

tube formation

in

changed

design.

lipoproteins.

levels of nitrate from 97r47 respectively

levels of malondlaldehyde

(MDA),

farnesyltransferase

Conclusions:

in medium

supplement or absence

atrovastatm,

I inhibitor,

Our results Identified

signaling pathway the s&tin-induced

on a matrix

gel.

supplemented

for 24 hours. of different

itavastatln

statin could explain, thrombotlc decrease.

events.

uM by 13+44% and from 1.4OkO.79 to 0.97iO.57

uM

and cerivastatin),

, and

(P=O.O73 and P=O.O09, respectwely)

but not

serum-induced endothelial tube formation. protein kinase) activity, but not the inhiblserum-Induced

which

inhibits

tube formation.

The addition

Ras farnesylation,

blocked

tube

simvastatin

Ras as a key player in the anti-angiogenic

at least in part. the protective which

action and

was greater

effect of these drugs against

than expected

simply

based

athero-

on the cholesterol

Angiotensin

MasakazuTanaka, Yamaguchi Clinical

experi-

kinds of reagents.

II Type-l Receptor Antagonist Inhibits Vascular Phenotypic Change and Remodeling in lntramyocardial Arteries by Reducing Oxidative Stress Through Upregulation of Superoxide Dismutase

1177-129

and

of free

with 5 %

In some

of statins. This is the first comprehensive analysis af the role of Ras in modulation of an angiogenic effect. The inhibition of angiogenesis by

to 90*52

(P~O.413 a marker

we

I” human

acid rescued simvastatin-Induced inhibition. Farnesylpyrophosphate, which Ras from the cytoplasm to the cell membrane, also blocked this inhibition. In

(FMD) to hyperemia from to’ 6.28+1.76 by 53*77%,

uM by 10*47%,

and by 15+45%, respectively

an in vitro model

serum

addition,

by 20*48%, respectively (P=O.O05 and P
signal

formation. RasN17 also inhibited serum-induced tube formation. Moreover, blocked epidermal growth factor-induced EGFP-R&NT translocation.

and improved the percent flow-mediated dilator response 4.57k1.63 to 5.96+1.88 by 38+40% and from 4.49+1.39 respectively

We developed

of mevalonic translocates

patients with coronary artery disease (CAD). 20 mg and placebo or ramipril 10 mg daily during

alone or combined

(statin).induced

tion of ~38 MAPK actwity, also suppressed

With

Shin, In S. Char, Eak K. Shm,

study was randomized, double-blind, placebo-controlled, ‘=P~0.05;~‘=Pc0.01;*“=Pc0.001 vs. Baseline. Data= mean+SD. Results:

and

Patients

of the biological

inhibitor therapies

Inhibitor

a hydrophilic statin (pravastatin), inhibited lnhibltion of p42/44 MAPK (mitogen-actwated

Enzyme

Disease

the mechanisms

enzyme

on

Keijiro Saku,

Japan

pEGFP(enhanced green fluorescent protein)-wild type (WT) Ras and pEGFP-dominant negative (N17) Ras expression vectors were constructed. Untransfected or transfected HCECs

in Hypercholesterolemic

Kwang K. Koh. Ji W. Son, Jeong Y. Ahn, Mi-Seunq Gachon Medical School, Incheon, South Korea

Fukuoka.

A reductase

Methods:

Function

Converting

*NO levels.

Background: While a recent report suggested that inhibition of Ras might be an effective anti-angiogenic therapy, it is unclear whether Ras mediates 3.hydroxy-3-methylglutatyl

SESSION

of Angiotensin

in serum

Shin-ichiroMiura MasahiroFujino, Yoshino Matsuo, Hiroaki Nishlkawa.

Results: Effects

with reductions

This

Tuesday, April 01, 2003, Noon9:OO p.m. McCormick Place, Hall A Presentation Hour: I:00 p.m.-2:OO p.m. Additive

associated

investigated that Ras plays a role in statin-mediated coronary artery endothelial cells (HCECs).

Drug Therapy and Endothelial

1177-126

arteries

Lipophilic Statin-Induced Antiangiogenesis Requires the inhibition of Ras Farnesylation in Human Coronary Artery Endothelial Cells

coenzyme

1177

brachial

ADR.

NS

were independent

that markers

in human

ESR measurements in aortic segments strongly suggest an eNOS origin for free radical production. These findings may have implications for cardiovascular complications noted

Seiji Umemoto,

University

Research

Graduate

Shinji Kawahara,

Kyoko Umeji, Masunori

School of Medicine,

Center, Yamaguchi

University

Matsuzaki,

Ube, Japan, Pharmaceutical

Hospital,

Ube, Japan

Background: Angiotensin II type-l receptor antagonists reduce reactive oxygen species (ROS) generated by activated NAD(P)H oxidase and vascular remodeling in hyperten-

PAI- antigen by -9*52% and by 11+38%, respectively (P=O.971 and P=O.O14, respectively). However, simvastatin combmed with ramlpril changed to greater extent FMD, and

slon, however,

plasma

We have examined whether Angiotensin II type-l receptor antagonists may inhibit vsscular remodeling and smooth muscle (SM) cell phenotypic change in intramyocardial artar-

levels of nitrate. MDA,

MCP-1,

CRP, and PAI-

Conclusions: Compared with slmvastatin additive effects on flow-mediated dilation inflammation

markers

and fibrinolysls

antigen

than simvastatin

alone.

alone, added ramipril to simvastatin showed and the plasma levels of nitrate and MDA,

potential

markers

in hypercholesterolemic

patients

with CAD.

the effect of ARAs on the scavenging

es via ROS-scavenging

In Vivo Evidence for Adriamycin-Induced Endothelial Nitric Oxide Synthase

27

Uncoupling of

University

of Michigan.

Background:

(ADR)

used anti-neoplastic

of a single

PNO) availability tion. Methods

dose of ADR is associated

and endothelial

and results:

nttrlc oxide synthase

A single dose of ADR (10 mgikg

agent

that IS well

with abnormalities

in nitric oxide

related superoxide

(O&

genera-

IV) in rabbits resulted I” reduced

peak relaxations to the agonists acetylcholine (Ach, 99&Z% to 57*4%) and A23187 (92*3% to 57+5%) and a shift in the ED, values. Peak relaxations to NTG were also altered (104+1 to 60*6) with no change I” ED, concentration. Exposure of aortic segments to ADR for 30 minutes resulted in dose dependent 9: generation, as measured by electron spin resonance (ESR) that was abolished by endothelial denudation and incubation

with diphenylene-iodinuim

(IOpM)

but not L-NMMA

(l@M).

Experiments

with

recombinant eNOS protein, conflrmed a dose dependent decrease in *NO generation as measured by L-Arginme-Citrulline assays and increase in 0,; production with ADR. Brachial artery flow medlated

dilation

(FMD) in patients

undergolng

ADR administration

attenuated after a single dose (6.5*1.0 to 2.5+1.1%, p=O.O004, 27+8 minutes). Serum nitrite and nitrate concentrations fell from

50+6 pmol/L

to 33r6

pre-ADR

pmol/L

of CD/131 thrombomodulin

pressure,

post ADR infusion

(p=O.O005)

and vWF activity remained

6 weeks

left ventricular

of treatment,

both drugs

hypertrophy

and fibrosis,

while serum

unchanged

and endothelial

showed

equipotent

synthetic-type

NO synthase

effects on blwd

myosln

expression

heavy

chain

in the heart. Furlher-

with the vehicle SHRSP group, E4177 showed a greater reduction of ratio in rntramyocardial arteries by reducing ROS assessed with iso-

and thiobarbituric

acid reactive

substances

by inhibiting

NAD(P)H

oxidase

essential subunit p22phox and upregulating CuiZn superoxide dismutase expression, and also inhibiting phenotypic modulation by reducing synthetic-type MHC NMHC-A and upregulating

contractile-type

pril. Conclusions:

Angiotensin

and phenotypic

change

SM-MHC II type-i

SM2 in the hearI more effectively

receptor

antagomsts

in the heart of SHRSP

than did cilaza-

may inhibit vascular

by reducing

remodeling

ROS via modulation

of the

expression of not only the generating enzymes but also the scavenging enzymes related to ROS more efficiently than ACE Inhibitor, and strategies almed at upregulating the scavenging

enzymes

in addition

ROS may have therapeutic

to downregulating

potential

the generating

against vascular

remodeling

enzymes

related

to

in hypertension.

Endothelial Function and Slow Coronary Flow

1177-130

Alps~ Sezqln, Ahmet Sigirci. lrfan Bar&u, Ergun Topal, Nurzen Sezgln, Ramazan Ozdemlr, Hakan Gullu, Ertan Yetkin, lzzet Tandogan, Now University, Malatya, Turkey, Baskent

University,

Adana,

Turkey

(60

“s/m*) was markedly time to end of infusion centrations

10 “g/kg/day)

Within

prostanes

IS a commonly

known to induce cardiotoxlclty. While the mechanisms of this effect are not entirely clear, evidence suggests that it is a free radical medlated process. We hypothewed that administration

Results:

more, compared the wall-to-lumen

Ann Arbor, Ml

Adriamycin

enzymes.

(MHC) NMHC-BiSMemb,

Damon Duquaine, Glenn Hirsch, Anjan Chakrabarti, B. Kalayanaraman, Joy Joseph, Christine Kehrer, Robert D. Brook, Jeanette Vasquez-Vivar, Sanjay Rajagopalan,

related to ROS is unclear.

Methods: Male stroke-prone SHR (SHRSP) were randomized and treated for 6 weeks with vehicle, AIIA (E4177; 30 “g/kg/day), or angiotensln-converting enzyme (ACE) inhibitor (cilazapril;

1177-l

enzymes

con-

after ADR

Objectives-The

aim of the study was to determine

SCF by using flow-mediated dilatation tecmque. Background- Slow coronary flow (SCF) in normal nized cllnical Methods

entity, but its etiopathogenesis

- Coronary flow was quantified

remans

endothelial coronary

function

angiogram

in patients with is a well-recog-

unclear.

using the corrected

Thrombosis

In Myocardial