777

Genistein inhibits the development of

atherosclerosis via inhibiting NF-kB and VCAM-1
expression in LDLR knockout mice
Juejin Wang, Rongjian Zhang, Youhua Xu, Hong Zhou, Bin Wang, and Shengnan Li

Abstract: Diet can be an important factor that influences risks for cardiovascular disease. Genistein (4’,5,7-
trihydroxyisoflavone), rich in soy, is one candidate that may benefit the cardiovascular system. Here, we explored the ef-
fect of genistein in atherosclerosis (AS) development in an in vivo mouse model. Low-density lipoprotein receptor
(LDLR) knockout mice were allocated to control, model, and genistein groups. Our results showed that genistein signifi-
cantly reduced the formation and development of atherosclerotic plaques ((4.68 ± 1.18) 106 versus (6.65 ± 1.51) 106
mm2, p < 0.05). In the genistein group, compared with the model group, total antioxidant capacity (TAC) level was 85.5 ±
15.6 versus 203.4 ± 32.6 mmol/L (p < 0.01); malondialdehyde (MDA) level was 3.79 ± 0.28 versus 3.06 ± 0.31 mmol/L
(p < 0.01), and superoxide dismutase (SOD) activity was 86.1 ± 6.1 versus 139.1 ± 25.1 U/mL (p < 0.01). Therefore, gen-
istein was able to enhance serum antioxidative ability in our mouse model. Genistein had no influence, however, on serum
cholesterol and lipid profiles. Genistein also markedly downregulated the expression of nuclear factor (NF)-kB and vascu-
lar cell adhesion molecule (VCAM)-1 in aortas of mice (p < 0.05). These observations suggest that genistein may inhibit
AS in LDLR–/– mice via enhancing serum antioxidation and downregulating NF-kB and VCAM-1 expression in the aorta.
Key words: genistein, atherosclerosis, NF-kB, VCAM-1, LDLR knockout mice.
Résumé : La diète peut être un facteur de risque important de maladies cardiovasculaires. La génistéine (4’,5,7-trihydro-
xyisoflavone), riche en soya, est un candidat qui pourrait être d’une grande utilité pour le système cardiovasculaire. Dans
le présent article, nous avons exploré l’effet de la génistéine sur le développement de l’athérosclérose (AS) dans des modè-
les in vivo. Nous avons réparti des souris au gène LDLR invalidé (knockout) dans trois groupes : témoin, modèle et génis-
téine. Nos résultats ont montré que la génistéine a diminué de façon significative la formation et le développement des
plaques d’athérosclérose ((4,6 ± 1,18)  10–6 contre (6,65 ± 1,51)  106 mm2, p < 0,05). Chez le groupe génistéine,
l’indice TAC a été de 85,5 ± 16,6 contre 203,4 ± 32,6 mmol/L (p < 0,01), le taux de MDA de 3,79 ± 0,28 contre 3,06 ±
0,31 mmol/L (p < 0,01) et l’activité SOD de 86,1 ± 6,1 contre 139,1 ± 25,1 U/mL (p < 0,01) par rapport au groupe mo-
dèle, respectivement. Ainsi, la génistéine a pu stimuler la capacité antioxydative sérique dans les modèles de souris. Toute-
fois, elle n’a pu influer sur les profils des lipides et du cholestérol sériques. La génistéine a aussi diminué
considérablement l’expression de NF-kB et de VCAM-1 dans les aortes de souris (p < 0,05). Ces observations donnent à
penser que la génistéine pourrait inhiber l’AS chez les souris LDLR–/– en stimulant l’antioxydation sérique et en diminuant
l’expression de NF-kB et de VCAM-1 dans l’aorte.
Mots-clés : génistéine, athérosclérose, NF-kB, VCAM-1, souris knockout LDLR.
[Traduit par la Rédaction]

Introduction 1998; Wong et al. 1998), inhibited vascular smooth muscle

Diet can be an important factor that influences the risks
for cardiovascular disease (Keys et al. 1986), and consump-
tion of a traditional Asian diet high in soy may play a key
role in preventing chronic diseases such as atherosclerosis
(AS) (Kolonel 1988). Among the active components of soy,
isoflavones appear to have an important role in preventing
cardiovascular disease. Previous studies showed that soy iso-
flavone decreased cholesterol concentration (Kirk et al.
Received 3 April 2008. Accepted 25 August 2008. Published on
the NRC Research Press Web site at cjpp.nrc.ca on 6 November
2008.
J. Wang, R. Zhang, Y. Xu, H. Zhou, B. Wang, and S. Li.1
Department of Pharmacology, Nanjing Medical University,
Hanzhong Road 140, Nanjing 210029, China.
1
Corresponding author (e-mail: snli@njmu.edu.cn).
cell (VSMC) proliferation (Dubey et al. 1999), improved
arterial compliance (Nestel et al. 1997), and had an anti-
oxidant action (Yamakoshi et al. 2000). Genistein (4’,5,7-
trihydroxyisoflavone), a dietary-derived isoflavonoid, has
various biological actions, including a weak estrogenic ef-
fect by binding to estrogen receptors (ERs) (Kuiper et
al.1998) and inhibition of protein tyrosine kinases (PTK)
(Nakashima et al. 1991). In experimental studies, genistein
significantly prevented thrombotic vessel occlusion (Kondo
et al. 2002).
AS is recognized as a chronic vascular inflammatory re-
sponse (Libby 2002), since inflammation is involved in all
stages of AS, from initiation through progression and, ulti-
mately, the thrombotic complications of AS (Hansson et al.
2006; Lucas et al. 2006). Many pathophysiologic compo-
nents, which are mediated by various proinflammatory me-
diators and cell adhesion molecules secreted by injured

Can. J. Physiol. Pharmacol. 86: 777–784 (2008) doi:10.1139/Y08-085 # 2008 NRC Canada
778
endothelial cells (ECs), such as nuclear factor kappa B
(NF-kB), vascular cell adhesion molecule-1 (VCAM-1),
thrombin, and tumor necrosis factor-a (TNF-a), play key
roles in the development of AS (Libby 2002; Pearson et al.
2003).
Emerging evidence indicates that genistein exerts multi-
faceted antiinflammatory effects in vasculature (Chacko et
al. 2005; Marotta et al. 2006), suggesting that genistein may
prevent AS by suppressing vascular inflammation. More-
over, the protective effects of soy isoflavones in AS
included a series of mechanisms such as prevention of low-
density lipoprotein (LDL) oxidation, improvement of vascu-
lar reactivity, inhibition of proinflammatory cytokines or
cell adhesion proteins, and reduction of platelet aggregation
(Wenzel et al. 2008). Recent studies have demonstrated that
genistein inhibits hyperpermeability of cultured vascular
ECs (Liu et al. 2005) and acts as antiinflammatory and anti-
atherogenic agents in human ECs (Simoncini et al. 2008).
Our previous data also showed that genistein inhibits
angiotensin-converting enzyme (ACE) (Xu et al. 2006),
which is a key enzyme in the development of AS (Lonn et
al. 2001). In particular, genistein was shown to be able to
inhibit NF-kB and VCAM-1 expression in vitro (Baxa and
Yoshimura 2003; Choi et al. 2003; Dijsselbloem et al.
2007; Mukherjee et al. 2003; Simoncini et al. 2008), which
is direct proof of an antiatherosclerotic effect of genistein.
Furthermore, previous observations indicated genistein or
isoflavone has an antiatherosclerotic effect in rabbits (Lee
et al. 2004; Yamakoshi et al. 2000); however, these mecha-
nisms of genistein are still not clarified in mice models in
vivo.
In the present study, we studied the effect of genistein on
atherogenesis in LDL receptor (LDLR) knockout mice. The
effect of genistein on lesion progression and plasma lipid
levels was observed. NF-kB and VCAM-1 expression in the
aortic atherosclerotic lesions was also analyzed in LDLR–/–
mice.
Materials and methods
Animals and treatment
Male LDLR knockout mice (Ishibashi et al. 1994) on
C57BL/6 background were obtained from the Model Animal
Research Center of Nanjing University. Experiments were
performed on age-matched mice at 2.5–4 months of age.
Mice in the control group (n = 6) were maintained on stand-
ard rat chow. Mice in the model and genistein experimental
groups were put on a western diet consisting of 21% fat by
weight, 0.15% cholesterol by weight, and no cholic acid
(made by the Model Animal Research Center). The experi-
mental genistein group (n = 6) was given genistein (Sigma,
St. Louis, USA) at 0.3 mg/kg in 100 mL of a mixture of
DMSO (1.25%) and PEG-400 (98.75%) s.c. daily. The
model group (n = 6) received genistein vehicle (100 mL of
a mixture of DMSO (1.25%) and PEG-400 (98.75%) s.c.
daily), and the control group was given placebo (normal sal-
ine) s.c. daily. Treatment lasted 8 weeks. Body weight and
food intake were monitored at regular intervals. The experi-
ments were conducted in accordance with the official rec-
ommendations of the Chinese Community Guidelines
concerning care and use of experimental animals.
Can. J. Physiol. Pharmacol. Vol. 86, 2008
Atherosclerotic lesion analysis
The proximal portion of the thoracic aorta up to the aortic
origin (aortic arch) was isolated and cleaned of the adherent
connective tissue, and samples were cut into 10-micrometre
sections. Five consecutive sections were taken at
120-micrometre intervals from each mouse. Sections were
fixed with 10% neutral buffered formalin for 30 min,
washed with PBS, and then treated with 60% isopropyl
alcohol for 1 min. The sections were then stained with
hematoxylin–eosin (HE). Images were captured with an
Olympus BX417 microscope and an Olympus digital
camera (Tokyo, Japan). The lipid-stained areas were ana-
lyzed with Adobe Photoshop software and Scion Image
software. The lipid-stained area was reported as the mean
area of the 5 sections from each mouse.
Plasma measurements
Mice were euthanized by CO2 inhalation. Blood was col-
lected from the retroorbital venous plexus. Plasma was sepa-
rated, and total cholesterol, triglycerides, LDL cholesterol,
and high-density lipoprotein (HDL) cholesterol were deter-
mined by using commercially available kits (Jiancheng Bio-
technology, Nanjing, China). Colorimetric changes were
measured at 500 nm. Total antioxidant capacity (TAC), ma-
londialdehyde (MDA), and superoxide dismutase (SOD)
were also measured by commercially available kits (Jian-
cheng Biotechnology, Nanjing, China).
Immunohistochemistry
To determine the expression of NF-kB and VCAM-1 in
the vessel wall of the LDLR knockout mice, cryosections
10 mm thick were fixed in acetone. In brief, the sections
were blocked in goat serum in calcium- and magnesium-
free PBS plus 0.005% Triton X-100 for 30 min. Tissue
sections were then incubated with anti-rabbit NF-kB p50
antibody or anti-rabbit VCAM-1 antibody (Santa Cruz Bio-
technology, Santa Cruz, USA) for 1 h at 37 8C. The areas
that were stained brown by anti-NF-kB p50 or anti-VCAM-1
antibodies were regarded as their respective localization and
were measured by the image analyzer. Quantitative analysis
was measured as a percentage of the positive-stained area in
the total atherosclerotic lesion area.
Tissue protein extraction and NF-kB p50 assay
Aortas were subjected to nuclear protein extraction for the
determination of activation of the p50 subunit, an index of
NF-kB activation (Borrás et al. 2005), following the instruc-
tions of the nuclear and cytoplasmic extraction kit (Pierce,
Rockford, USA). The efficiency of extraction was deter-
mined by measuring the relative activity of a cytosolic en-
zyme (lactate dehydrogenase) in tissues. The active form of
the p50 subunit was then detected in extracts by ELISA ac-
cording to the manufacturer’s instructions (TransAM NF-kB
p50 Chemi, Active Motif, Carlsbad, USA).
Reverse transcription (RT)-PCR amplification
The aortas of mice were dissected and kept in liquid ni-
trogen. Total RNA was then extracted from the aortas by us-
ing Trizol reagent (Invitrogen, Carlsbad, USA). Two
hundred microliters of chloroform were added to the homo-
genates, and these were then incubated for 10 min at room
# 2008 NRC Canada
Wang et al. 779

Fig. 1. Representative micrographs showing HE-stained lesions

(A) control, (B) model, and (C) genistein LDLR–/– mice
(magnification  400). (D) Lesion size in the aortic arch stained by
HE in LDLR–/– mice. Data are means ± SD. **, Significant at p <
0.01 vs. control; #, p < 0.05 vs. model, n = 6 mice per group, 8 sec-
tions per mouse. HE, hematoxylin–eosin; LDLR, low-density lipo-
protein receptor.

temperature. After centrifugation at 12 000g for 10 min,

500 mL of isopropyl alcohol was added. The samples were
then incubated for 10 min at room temperature and centri-
fuged at 12 000g for 5 min. RNA pellets were washed with
75% ethanol and dried. RT was performed with oligo(dT).
The following oligonucleotide primer pairs were examined:
b-actin sense, 5’-GAC GGC CAG GTC ATC ACT AT-3’,
and antisense, 5’-CTT CTG CAT CCT GTC AGC AA-3’;
VCAM-1 sense, 5’-CGT CGC GAG GTT GTT TAG AGT-
3’, and antisense, 5’-CAA CAG TCA GTC CAA GCA ACA
CT-3’. PCR assays were performed for 35 cycles. Each
cycle consisted of the following steps: denaturation at 94 8C
for 30 s, annealing at 56 8C for 45 s, and extension at 72 8C
for 1 min. PCR products were analyzed in 2% agarose gel
containing ethidium bromide. Each mRNA level was ex-
pressed as a ratio to b-actin mRNA.

Statistical analysis
The data were represented as means ± SD. The compari-
son of more than 2 groups was determined by one-way
ANOVA, followed by Bonferroni test as appropriate, using
Stata 7.0 (Stata, College Station, USA). Statistical signifi-
cance was considered at p < 0.05.

Results
Reduced atherosclerotic lesions in genistein group mice
To examine the effect of genistein in vivo, genistein was
administrated to mice for 8 weeks. As a result, genistein sig-
nificantly reduced the area of atherosclerotic lesions in this
group. Figure 1 shows examples of the different stages of
AS lesion development observed on examination by light
microscope; sections of aorta also revealed significant dif-
ferences in lesion area between the genistein and model
groups. Counting by 8 consecutive sections occupied by
HE-stained areas, lesions were (6.65 ± 1.51) 106 mm2 in
the model group versus (4.68 ± 1.18) 106 mm2 in the gen-
istein group (p < 0.05).

Food intake, body weight, and plasma cholesterol and

lipid profiles
After 8 weeks of treatment, food intake and body weight
did not differ among the 3 groups (data not shown).
Although serum cholesterol and lipid profiles play an impor-
tant role in the development of AS, there is disagreement on
the effect of genistein on plasma cholesterol and lipid pro-
files. In this study, genistein had no effect on the plasma
lipid profile, which is consistent with previous reports
(Atteritano et al. 2007; Nagarajan et al. 2008). Table 1
shows that plasma total cholesterol, HDL cholesterol, LDL
cholesterol, and triglycerides were not different from those
of the model group after chronic treatment with genistein.
# 2008 NRC Canada
780 Can. J. Physiol. Pharmacol. Vol. 86, 2008

Table 1. Plasma lipid levels (mmol/L) in LDLR–/– mice main- Fig. 2. (A) TAC level in the presence or absence of genistein.
tained for 8 weeks on a high-fat diet in the presence or absence of (B) MDA level in the presence or absence of genistein. (C) SOD
genistein. activity in the presence or absence of genistein in LDLR–/– mice.
Data are means ± SD. *, Significant at p < 0.05 and **, p < 0.01
Control Model Genistein vs. control; ##, p < 0.01 vs. model, n = 6 per group. TAC, total
Total cholesterol 8.8±1.8 16.7±2.1** 15.9±1.1** antioxidant capacity; MDA, malondialdehyde; SOD, superoxide
HDL cholesterol 0.48±0.06 0.62±0.13* 0.68±0.07** dismutase.
LDL cholesterol 10.2±1.3 12.9±1.4** 13.2±1.2**
Triglycerides 0.39±0.05 0.58±0.07** 0.62±0.04**
Note: LDLR–/–, low-density lipoprotein receptor knockout. Control mice
received a standard diet. Model (vehicle) and genistein groups received a
high-fat diet. Data are means ± SD. *, Significant at p < 0.05 and **, p <
0.01 vs. control, n = 6 per group.

Plasma TAC, MDA, and SOD levels

To examine the antioxidant ability of genistein in vivo,
plasma of mice was separated to examine TAC, MDA, and
SOD levels. Compared with the model group, TAC level in
the genistein group was 85.5 ± 15.6 versus 203.4 ±
32.6 mmol/L (p < 0.01) (Fig. 2A), MDA level was 3.79 ±
0.28 versus 3.06 ± 0.31 mmol/L (p < 0.01) (Fig. 2B), and
SOD activity was 86.1 ± 6.1 versus 139.1 ± 25.1 U/mL
(p < 0.01) (Fig. 2C).

Expression of NF-kB and VCAM-1 in aorta

To investigate the mechanism of genistein’s effect on the
development of AS, we next examined the expression of
NF-kB p50 and VCAM-1. There was minimal NF-kB p50
reaction produced in the ECs of aorta in control animals.
But in model animals, the localization of NF-kB p50 was
observed in ECs of aorta, even in VSMCs. Genistein de-
creased the localization of NF-kB p50 reaction production.
As shown in Fig. 3, limited NF-kB reaction production was
observed in ECs of aorta. In addition, treatment with genis-
tein also decreased the level of the p50 subunit of NF-kB in
lysis derived from aorta (p < 0.05) (Fig. 3E).
There was minimal VCAM-1 expression in control
LDLR–/– mice. The immunostaining of this protein became
obvious, however, in the model LDLR–/– mice. Genistein
markedly reduced immunoactive VCAM-1 expression (p <
0.05) (Fig. 4). The level of VCAM-1 mRNA in LDLR–/–
mice treated with genistein (0.28 ± 0.13) was also signifi-
cantly reduced compared with that of the model group
(0.75 ± 0.15) (p < 0.01) (Fig. 5). These observations indi-
cated that genistein inhibited the development of AS, which
may be attributed to the suppression of NF-kB and VCAM-1
expression in the aorta.
During the past decade, extensive studies have largely fo-
Discussion cused on elucidating the effect of genistein on lipid profiles
Genistein is known to have a number of antiatherogenic because hyperlipidemia contributes to AS (Vitolins et al.
actions, as described in the Introduction. In vitro, genistein 2001). In our study, however, there were no differences be-
inhibited the migration and proliferation of SMCs (Pan et tween the model and genistein groups on the plasma lipids
al. 2001; Shimokado et al. 1994), which is important in pro- and lipoproteins (Table 1). This result is consistent with pre-
motion and progression of the atherosclerotic process. Geni- vious reports. Atteritano et al. (2007) showed that genistein
stein may also suppress thrombus formation by inhibiting administration did not affect blood lipid levels in postmeno-
platelet activation (Kuruvilla et al. 1993; Murphy et al. pausal women; Nagarajan et al. (2008) also indicated that
1993) and aggregation (Asahi et al. 1992; Kondo et al. feeding soy protein isolate (SPI) in mice did not affect
2002) and by reducing platelet serotonin uptake (Helmeste plasma concentrations of HDL, LDL, and total cholesterol
and Tang 1995). In the present study, our in vivo data indi- or triglycerides, suggesting that the antiatherogenic effect of
cated that genistein could decrease the area of atheroscler- genistein is not due to a change in the plasma lipids.
otic lesion in LDLR–/– mice (Fig. 1). Lipid peroxidation due to a hyperlipidemic diet may be
# 2008 NRC Canada
Wang et al. 781

Fig. 3. Immunohistochemical staining for NF-kB p50 in the aorta of LDLR–/– mice, (A) control, (B) model, and (C) genistein
(magnification  400). (D) Comparison of the NF-kB p50 protein expression among groups in LDLR–/–mice. Data are means ± SD. **, Sig-
nificant at p < 0.01 vs. control; ##, p < 0.01 vs. model, n = 6 per group. (E) Levels of the active p50 subunit of NF-kB were measured in
lysis from aorta treated with genistein. Data are means ± SD for 6 different experiments. *, Significant at p < 0.05 vs. control; #, p < 0.05
vs. model. NF-kB, nuclear factor kB; LDLR, low-density lipoprotein receptor.

the predominant factor of endothelial injury in the LDLR–/–
mice model. Genistein has antiatherogenic effects, one po-
tential mechanism of which is that genistein has antioxidant
properties, including scavenging of hydrogen peroxide and
superoxide anion, in mRNA and protein levels (Borrás et al.
2006; Wei et al. 1993). We further examined the oxidative
indexes of plasma in the present study. Our data demon-
strated that genistein enhanced TAC and SOD levels in
plasma while reducing MDA level (Fig. 2), suggesting that
the inhibition of the atherosclerotic lesion by genistein may
be due to the improvement of endothelial dysfunction via its
antioxidative ability.
NF-kB is one of the first mediators activated in the in-
flammatory response that plays a major role in atherogene-
sis, and its activation has been reported in atherosclerotic
plaques and in cultured cells (Collins and Cybulsky 2001;
Kutuk and Basaga 2003). Clearly, inhibition of NF-kB could
attenuate the development of AS in apoE/LDLR double-
knockout mice (Jawień et al. 2005). It was shown in
previous reports that genistein can inhibit the activation of
NF-kB in human cancer cells (prostate, pancreatic, and
breast) (Gong et al. 2003; Li et al. 2004; Raffoul et al.
2006). Bhullar et al. (1998) indicated that genistein inhibited
NF-kB by attenuating shear stress-induced IkB kinase via
phosphorylation and subsequent degradation of IkB protein
in bovine aortic endothelial cells (BAECs). Futhermore, it
was also shown that genistein could inhibit the expression
of NF-kB in dendritic cells, macrophages cells, and T lym-
phoma cells (Baxa and Yoshimura 2003; Choi et al. 2003;
Dijsselbloem et al. 2007), which play important roles in in-
# 2008 NRC Canada
782
Fig. 4. Immunohistochemical staining for VCAM-1 in the aorta of
LDLR–/– mice (A) control, (B) model, and (C) genistein
(magnification  400). (D) Comparison of the VCAM-1 protein ex-
pression among groups in LDLR–/– mice. *, Significant at p < 0.05
and **, p < 0.01 vs. control; ##, p < 0.01 vs. model, n = 6 per
group.
Can. J. Physiol. Pharmacol. Vol. 86, 2008
Fig. 5. RT-PCR analysis of VCAM-1 expressed in the aorta of
LDLR–/– mice (A) control, (B) model, and (C) genistein. (D) Com-
parison of the mRNA level of VCAM-1 among groups in LDLR–/–
mice. Data are relative intensity versus b-actin mRNA (mean ±
SD). **, Significant at p < 0.01 vs. control; #, p < 0.05 vs. model,
n = 6 per group. VCAM, vascular cell adhesion molecule;
LDLR, low-density lipoprotein receptor.
flammatory responses. Taken together, these observations
showed that genistein could inhibit the expression of NF-kB
not only in ECs but also in inflammatory cells in vitro.
Therefore, we further investigated genistein’s influence on
NF-kB levels in vivo. Our data showed that genistein signif-
icantly decreased the protein expression of NF-kB p50, an
index of NF-kB activation, in atherosclerotic aortas (Fig. 3),
suggesting that genistein may suppress the development of
AS via inhibiting NF-kB activation in LDLR–/– mice.
Moreover, we found that genistein treatment led to a sig-
nificant reduction in the adhesion molecule VCAM-1. The
adhesion of circulating leukocytes to vascular ECs plays an
important role in the early stages of AS (Lombardini et al.
1997). The early steps of AS appear to be mediated through
a diverse family of cellular adhesion molecules that are ex-
pressed on the surface of vascular ECs (Price and Loscalzo
1999). Among the identified adhesion molecules involved in
the atherosclerotic process, the expression and biologic
properties of VCAM-1 are well characterized (Blankenberg
# 2008 NRC Canada
Wang et al.
et al. 2003). Previous reports showed that plasma soluble
VCAM-1 was significantly reduced by genistein administra-
tion in postmenopausal women (Atteritano et al. 2007;
Colacurci et al. 2005). It is also indicated that phenoxodiol,
a synthetic analogue of genistein, inhibited VCAM-1 expres-
sion in vivo (Gamble et al. 2006). Furthermore, genistein at
low concentrations suppressed TNF-a-induced VCAM-1
mRNA expression in human umbilical vein endothelial cells
(HUVECs) (Mukherjee et al. 2003). Finally, Simoncini et al.
(2008) indicated that red clover extracts, particularly genis-
tein and daidzein, inhibited the endothelial expression of
VCAM-1 induced by bacterial lipopolysaccharide, acting as
antiinflammatory and antiatherogenic agents on human ECs.
Our data also showed that genistein decreased VCAM-1 pro-
tein and VCAM-1 mRNA expression in the aorta of
LDLR–/– mice (Figs. 4 and 5). These results reasonably im-
plicate the decreased expression of VCAM-1 protein and
mRNA in the aorta in genistein’s inhibition of the develop-
ment of AS.
In conclusion, genistein inhibited the progression of AS in
LDLR–/– mice. The mechanisms may include its antioxida-
tive action and its effect in decreasing NF-kB and VCAM-1
expression in the aorta, rather than its direct action on blood
lipid concentration. Further studies are needed for under-
standing other mechanisms of genistein’s antiatherogenic ac-
tion in detail.
Acknowledgements
This work was supported by Key Project of Jiangsu Pro-
vincial Natural Scientific Fund (No. BK2006727).
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