Embed Size (px)
Citation preview
Effect of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucosideon lipoprotein oxidation and proliferation of coronary
arterial smooth cells
QI-LI LIU, JUN-HUA XIAO, RONG MA, YI BAN and JIA-LING WANG*
Department of Pharmacology, Tongji Medical college of Huazhong University of Science andTechnology, Wuhan 430030, China
(Received 6 January 2005; revised 14 April 2005; in final form 2 May 2005)
To investigate the effects of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside (THSG), a compoundextracted from the root of Polygonum multiflorum Thunb, on lipoprotein (LDL and VLDL) oxidation,proliferation and nitric oxide (NO) content of coronary arterial smooth cells (CASMCs) induced by LDL,VLDL, ox-LDL and ox-VLDL. The oxidation level of lipoprotein was determined by thiobabituric acid(TBA) method and agarose gel electrophoresis. The proliferative degree was determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) method. The NO content was assayed bynitrate reductase method. (1)THSG (0.1 , 100mmol·L21) could dose-dependently prevent lipoproteinfrom oxidation induced by Cu2þ and CASMCs. (2)THSG (0.1 , 100mmol·L21) inhibited porcineCASMCs proliferation elicited by LDL, VLDL, ox-LDL and ox-VLDL. (3)THSG(0.1 , 100mmol·L21) counterpoised the decrease of NO content in CASMCs evoked by LDL,VLDL, ox-LDL and ox-VLDL. As compared with control, it was found that the threshold concentrationof THSG, which significantly exerted the actions mentioned above, were at the concentration of1mmol·L21 (P , 0.01). In conclusion, THSG possesses the antagonistic effects on oxidation oflipoprotein, proliferation and decrease of NO content of CASMCs, which partially explain themechanism of anti-atherosclerosis of Polygonum multiflorum Thunb.
Keywords: 2,3,5,40-Tetrahydroxystilbene-2-O-beta-D-glucoside; Lipoprotein oxidation; Proliferation;Coronary arterial smooth cells
1. Introduction
Atherosclerosis (AS) is one of the most common diseases. The proliferation of vascular
smooth muscle cells (SMC) and oxidative modification of lipoprotein are critical events in
the AS formation. Low density lipoprotein (LDL) and very low-density lipoprotein (VLDL),
particularly oxidated lipoprotein (ox-LDL, ox-VLDL) are regarded as the major risk factors
of AS [1,2]. Moreover, the decrease of nitric oxide (NO) level in serum could induce the
SMC proliferation and this response elicits the development of atherosclerotic lesions [3].
Polygonum multiflorum Thunb, a traditional Chinese medicine, possesses extensive
pharmacological effects. It has an obvious anti-atherosclerotic effect, effectively decreases
Journal of Asian Natural Products Research
ISSN 1028-6020 print/ISSN 1477-2213 online q 2007 Taylor & Francis
http://www.tandf.co.uk/journals
DOI: 10.1080/17415990500209064
*Corresponding author. Email: [email protected]
Journal of Asian Natural Products Research, Vol. 9, No. 5, July–August 2007, 421–429
the total cholesterol in human and rat serum, strongly suppresses the oxidation, and cleans
the active oxygen free radicals [4]. It has been reported that Polygonum multiflorum Thunb
has antiproliferative capability to vascular smooth muscle cell (VSMC), increases the
content of NO in VSMC culture medium, and inhibits the decrease of NO in high lipoprotein
medium for VSMC culture.
Generally, 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside (THSG), isolated from the
roots of Polygonum multiflorum Thunb, is regarded as an active compound. The chemical
structure (shown in figure 1) of the aglycone part of THSG is very similar to 3,5,40-
trihydroxystilbene (resveratrol) and even to diethylstilbestrol. Resveratrol and diethyl-
stilbestrol belong to hydroxystilbene compounds, and exert the antagonistic effect on LDL
and VLDL oxidative modification, eliciting the synthesis and release of NO, antagonizing
the SMC proliferation induced by lipoprotein and its oxidant [5–7]. So we wonder whether
THSG has the same action as that of the roots of Polygonum multiflorum Thunb, mentioned
above.
This paper firstly deals with the antioxidative effect and antiproliferative effect of THSG
on porcine CASMCs and its relative mechanisms.
2. Results and discussion
2.1 Results
2.1.1 Assay of THSG. By screening, the HPLC performance condition, to employ
Lichrospher 5-C18 (5mm, 4.6 £ 250 mm) as the stationary phase, methanol–water (4:6) as
mobile phase, 310 nm as the detective wavelength, was determined. Within the range from
0.14 to 2.40mg/20mL, there was a good linearity with the regression equation of
Y ¼ 2.84 £ 1027X þ 0.0123, r ¼ 0.9995, the average recovery was 101.4%, RSD was
1.64% (n ¼ 6). As shown in figure 2, the content of THSG was up to 99.04%, which tallied
with the monomer quality standard.
2.1.2 Effect of THSG on oxidative modification of VLDL and LDL induced by Cu21 .
As shown in table 1 and figure 3, after incubation for 24 h at 378C, MDA content was
evaluated by the TBA method (using the MDA kit) and the oxidation level of lipoprotein was
determined by 1% agarose gel electrophoresis. THSG of 0.1 , 100mmol·L21 could
concentration-dependently prevent the LDL and VLDL from lipoprotein oxidation induced
Figure 1. The chemical structure of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside (THSG).
Q.-L. Liu et al.422
by Cu2þ , and the effects of anti-oxidation of THSG on LDL and VLDL were much stronger
than those of vitamin C.
2.1.3 Effect of THSG on oxidative modificaton of LDL and VLDL induced by porcine
CASMCs. As shown in table 2, porcine CASMCs could significantly induced LDL and
VLDL oxidation (P , 0.01) too. THSG of 0.1 , 100mmol·L21 could concentration-
dependently inhibited this LDL and VLDL oxidation induced by porcine CASMCs.
Table 1. The inhibitory effect of THGS on lipoprotein (LDL and VLDL) oxidation induced by Cu2þ .
MDA content of culture medium (nmol·mgprot21)
Concentration of drug (mmol·L21) VLDL LDL
Blank control 43.40 ^ 6.75 35.71 ^ 4.41Positive control 220.47 ^ 19.08## 137.50 ^ 9.16##
Vitamin C 150 185.76 ^ 23.86** 111.31 ^ 13.57**THSG 0.1 218.17 ^ 17.31 132.74 ^ 6.20THSG 1 185.19 ^ 13.09** 113.10 ^ 11.31*THSG 10 160.88 ^ 10.43** 102.38 ^ 8.05**THSG 100 85.07 ^ 7.60** 78.57 ^ 6.27**
n ¼ 8, x ^ s * P , 0.05, vs positive control group, ** P , 0.01, vs positive control group,## P , 0.01, vs blank control group.
Figure 2. HPLC chromatograms of THSG standard (the left) and self-prepared THSG (the right).
Figure 3. The inhibitiory effect of THSG on LDL oxidation induced by Cu2þ (One case) Form right to left: 7: LDLuntreated with Cuþþ , 6: LDL or VLDL treated with Cuþþ , 5: treated with Vitamin C, and 4 , 1: treated withTHGS 0.1, 1, 10 or 100mmol·L21 respectively.
Effect of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside 423
2.1.4 Effect of THSG on the proliferation of porcine CASMCs induced by LDL, VLDL,
ox-LDL and ox-VLDL. LDL, VLDL, ox-LDL, or ox-VLDL could remarkably accelerate
the proliferation of porcine CASMCs (P , 0.01). But this proliferation could be
concentration-dependently inhibited by THSG (0.1 , 100mmol·L21). THSG at low
concentration of 1mmol·L21 had obvious anti-proliferation effect (P , 0.05) and THSG at
high concentration of 100mmol·L21 could almost completely antagonize the proliferation of
CASMCs induced by LDL and VLDL, shown in table 3.
2.1.5 Influence of THSG on the decrease of NO content in the medium of cultured
porcine CASMCs Induced by LDL, ox-LDL, VLDL or ox-VLDL. As shown in table 4,
Table 2. The inhibitory effect of THGS on lipoprotein (LDL and VLDL) oxidation induced by CASMCs.
MDA content of culture medium (nmol·mgprot21)
Concentration of drug (mmol·L21) VLDL LDL
Blank control 44.73 ^ 6.52 41.85 ^ 6.52Positive control 98.12 ^ 10.49## 66.38 ^ 4.47##
Vitamin C 150 76.48 ^ 8.51** 54.83 ^ 7.07*THSG 0.1 89.47 ^ 7.07 60.61 ^ 7.74THSG 1 82.25 ^ 4.74** 57.72 ^ 7.07*THSG 10 72.15 ^ 8.94** 51.95 ^ 5.48*THSG 100 57.72 ^ 8.94** 44.73 ^ 6.52**
n ¼ 6, x ^ s * P , 0.05, vs positive control group, ** P , 0.01, vs positive control group, ## P , 0.01, vs blank control group.
Table 3. The inhibitory effect of THGS on proliferation of cultured porcine CASMCs induced by LDL, VLDL, ox-LDL and ox-VLDL.
Value of OD
Concentration of drug(mmol·L21) VLDL ox-VLDL LDL ox-LDL
Blank control 0.2022 ^ 0.015 0.2024 ^ 0.017 0.2129 ^ 0.007 0.2094 ^ 0.010Positive control 0.2440 ^ 0.012## 0.2678 ^ 0.015## 0.2391 ^ 0.014## 0.2599 ^ 0.017##
Vitamin C 150 0.2265 ^ 0.010** 0.2505 ^ 0.009* 0.2202 ^ 0.011** 0.2412 ^ 0.010*THSG 0.1 0.2350 ^ 0.013 0.2627 ^ 0.016 0.2312 ^ 0.013 0.2513 ^ 0.016THSG 1 0.2292 ^ 0.011* 0.2569 ^ 0.013* 0.2238 ^ 0.011* 0.2451 ^ 0.013THSG 10 0.2238 ^ 0.012* 0.2487 ^ 0.016** 0.2179 ^ 0.010** 0.2341 ^ 0.011*THSG 100 0.2068 ^ 0.016** 0.2382 ^ 0.009** 0.2105 ^ 0.008** 0.2271 ^ 0.009**
n ¼ 8, x ^ s. * P , 0.05, vs positive control group, ** P , 0.01, vs positive control group, ## P , 0.01, vs blank control group.
Table 4. Effects of THGS on NO content and decrease of porcrine CASMCs induced by LDL, VLDL, ox-LDL andox-VLDL.
NO content of culture medium (mmol·L21)
Concentration of drug (mmol·L21) VLDL ox-VLDL LDL ox-LDL
Blank control 60.90 ^ 4.70 59.46 ^ 4.02 64.77 ^ 5.04 64.01 ^ 5.42Positive control 35.15 ^ 5.43## 34.31 ^ 2.88## 47.66 ^ 5.22## 42.46 ^ 1.74##
Vitamin C 150 50.60 ^ 6.57** 45.24 ^ 9.83** 56.80 ^ 4.02** 51.91 ^ 5.11**THSG 0.1 39.52 ^ 3.43 36.19 ^ 5.76 48.87 ^ 3.88 42.32 ^ 5.10THSG 1 48.47 ^ 5.59** 46.22 ^ 5.73** 56.27 ^ 4.19** 52.05 ^ 6.05**THSG 10 59.39 ^ 3.78** 52.71 ^ 7.05** 63.68 ^ 3.99** 60.17 ^ 4.79**THSG 100 69.02 ^ 5.33**
# 63.59 ^ 5.24** 72.25 ^ 5.61** 65.55 ^ 6.98**
n ¼ 6, x ^ s. * P , 0.05, vs positive control group, ** P , 0.01, vs positive control group, # P , 0.01, vs blank control group, ##
P , 0.01, vs blank control group.
Q.-L. Liu et al.424
LDL, VLDL, ox-LDL, and ox-VLDL could significantly reduce the NO content produced by
CASMCs (P , 0.01). THSG at concentration from 0.1 to 100mmol·L21 could inhibit the
decrease of NO content induced by LDL, VLDL, ox-LDL and ox-VLDL, in a concentration-
dependent manner. THSG 1mmol·L21 had obvious effect and its efficacy is similar
to that of vitamin C 150mmol·L21, and THSG at high concentration of 100mmol·L21
could completely inhibit, even inverse the decrease of NO content induced by LDL, VLDL,
ox-LDL and ox-VLDL.
2.2 Discussion
2.2.1 Possible mechanism of inhibiting atherosclerosis formation of THSG. The
development of atherosclerosis (AS) is promoted by many factors, which include the
oxidative modification of lipoprotein [8], the decrease of NO synthesis or/and NO release in
SMCs, and the proliferation of SMCs. Coronary events are closely related to the progression
of AS [8]. The proliferation of SMCs is one of the main pathological changes from
metaphase to late AS. LDL can cause the elevation of asymmetrical dimethylarginine
(ADMA) [9], which is associated with the decrease of NO production and the impairment of
endothelium-dependent vasodilator function. Moreover, various matters can modify LDL
(VLDL) into ox-LDL (ox-VLDL) in different fashions. Ox-LDL (ox-VLDL) not only
promotes the formation of foam cells in the artery wall, but also consumingly stimulates the
SMC proliferation [10,11]. In addition, ox-LDL can reduce the synthesis and release of NO
by inhibiting inducible nitric oxide synthesis (iNOS) in macrophages and VSMC [12]. So in
this paper, we studied the inhibitory effects of THSG on the oxidative modification of
lipoprotein, the CASMCs proliferation and the synthesis and release of NO in order to
explore its mechanism of antagonizing AS.
We found THSG (0.1 , 100mmol·L21) could dose-dependently prevent LDL and
VLDL from oxidation induced by Cu2þ and CASMCs. These results suggest that the
anti-atherosclerosis effect of THSG maybe partly due to inhibiting lipoprotein oxidation.
Furthermore, compared with the control group, THSG from 0.1 to 100mmol·L21 could
significantly antagonize the decrease of NO synthesis and NO release mediated by
lipoproteins and those oxidative modifications. In the same experiment, THSG at 10,
100mmol·L21 almost completely inhibited the decrease of NO, and even increased the
content of NO in the medium. These results suggest that THSG could promote the
synthesis of NO in SMC or the release from SMC, which is consistent with our previous
result: THSG is an elicitor of the nitric oxide synthesis [13]. Lipoprotein (especially their
oxidation) and reduction of NO content can promote SMC proliferation [14] and
accelerate the development and formation of atherosclerotic lesion [3]. The anti-
proliferative effect of NO on SMC is mediated via cGMP-dependent activation of the
cAMP kinase mechanism [15]. Our results also proved THSG (0.1 , 100mmol·L21)
could inhibit porcine CASMCs proliferation induced by lipoproteins and those oxidative
modifications.
In conclusion, THSG is not only an inhibitor of LDL and VLDL oxidation and CASMCs
proliferation, but also a promoter of the NO production of CASMCs.
2.2.2 The potencies comparisons between THSG and others. Vitamin C (ascorbic acid) is
one of the most effective water–soluble antioxidant in human Plasma [16] and has been
Effect of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside 425
widely used as a conventional medicine in AS treatment. So it was chosen as positive drug in
our experiment. Our results show the effects (potencies) of vitamin C are weaker than those
of THSG.
Resveratrol may be an anti-atherosclerosis agent [5,17]. Therefore, we made a comparison
between the resveratrol’s potency from reference [17] and that of THSG from our data, and it
was found that the former maybe not stronger than the latter. Generally, stilbene compounds,
particularly, diethylstilbestrol compounds are considered possessing estrogen-like action.
Therefore, they have not been used as a choice for AS treatment. Resveratrol has a weak
estrogen-like action but has a strong suppression on AS, so it could be explored as a valuable
drug for clinic use. Our study demonstrated that THSG also had a weak estrogen-like action,
which equaled to 10 percent of estrogen effect (unpublished data), so we also consider THSG
could be explored as a valuable drug.
3. Experiment
3.1 Drug and agents
Polygonum multiflorum Thunb was purchased from DeQing county, GuangDong province
and identified by Wang XiaoMin, Chief Pharmaceutist. THSG [18], a main water–soluble
compound, was extracted from Polygonum multiflorum Thunb, isolated by conventional
methods and provided by phytochemistry Lab of pharmacological department, Tongji
Medical College. The content of THSG was assayed through Waters high performance liquid
chromatography (HPLC) system, including Waters 600 controller, Waters Delta 600 and
Waters 2996 Photodiode Array, manipulated by Millennium software. The monomer quality
standard THSG was purchased from the center of standard compounds of national food and
drug administer.
Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS) and trypsin
were purchased from GIBCO. 1,1,3,3,-tetramethosy-propane (TMP) and 3-[4,5-
dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) were purchased from
Sigma Chemical CO. Malondialdehyde (MDA) test kit and NO test kit were the products
of Nanjing Jianchen Company. Vitamin C (ascorbic acid) was purchased from Dongbei
pharmaceutical CO. Agarose and Sudan black B were obtained from Biowest CO. and
Chroma CO., respectively.
3.2 Porcine coronary arterial smooth muscle cells (CASMCs) isolation
Porcrine CASMCs were isolated with the method as description in reference [19].
The intima was first peeled off from the artery, then the media carefully stripped away
from the adventitia and placed in a Petri dish containing warmed HEPES-buffered DMEM
(378C). The medial layer was cut into approximately 1 mm2 squares, which were
transferred to a 25 cm2 tissue culture flask and barely covered with DMEM supplemented
with 20% FBS. The blocks of tissue were cultured at 378C in a humid atmosphere of 5%
CO2 and 95% air (vol/vol). After 3 , 4 weeks, the tissue blocks were removed and
the migrated CASMCs were cultured, followed by subculture using trypsinization.
The identity of the CASMCs was confirmed by morphological examination and by
staining for a-actin.
Q.-L. Liu et al.426
3.3 Isolation and preparation of lipoproteins
VLDL and LDL were isolated from fresh healthy human plasma according to the method
described by reference [20]. Adjusting plasma (without chylimicron) density to 1.20 with
solid NaBr, then the mixture of LDL and VLDL (top-layer) was isolated from the plasma by
density-gradient ultracentrifugation at 60 000 rpm for 5 h at 108C (a Beckman 70.1 Tirotor in
a Beckman L8-80 ultracentrifuge). VLDL (density 1.006) and LDL (density 1.063) were
isolated from mixture of LDL and VLDL by density-gradient ultracentrifugation, in NaBr
solution at density 1.006 and 1.063 respectively, at 60 000 rpm for 5 h, at 108C. Isolated
fractions were dialyzed at 48C against phosphate-buffered saline (PBS) in 6000–8000
molecular weight cut off dialysis membrane for 24 h, changing the PBS every hour, and then
concentrated using polyethylene glycol (20 000 molecular weight) concentrator with 10 kDa
cut off membrane under nitrogen.
3.4 VLDL and LDL oxidative modification and oxidation level evaluation
The protein content of the LDL and VLDL were determined by a modified Lowry assay.
Oxidative reaction of LDL and VLDL were induced by CuSO4 with final concentration was
10mmol·L21, for 24 h. The reaction was stopped by 2 mmol·L21 of ethylenediamine
tetraacetic acid (EDTA) and 20mmol·L21 of butylated hydroxytoluene. VLDL and LDL
were dialyzed at 48C against phosphate-buffered saline (PBS) in 6000–8000 molecular
weight cut off dialysis membrane for 24 h (change the PBS every hours) for cleaning the
Cu2þ . The lipid hydroperoxide contents of the oxidative modification of LDL and VLDL
were assayed by a thiobabituric acid (TBA) method and the relative electrophoretic mobility
compared with normal LDL and VLDL, as an index of protein modification, were
determined by agarose gel electrophoresis.
3.5 Cell viability assay
Mitochondrial dehydrogenase activity, as an index of cell viability, was assessed using the
MTT assay. The porcine CASMCs were incubated with defined lipoprotein (LDL, VLDL,
ox-LDL, and ox-VLDL) and drug treatment for 20 h. Culture medium was then replaced with
fresh medium containing MTT (500mg·mL), and cells were incubated for an additional 4 h.
Then, dimethyl sulphoxide (DMSO) was added to each well, and the microtiter plates were
incubated at room temperature for another 20 minutes. The absorbance was measured at
570 nm through a microplate reader.
3.6 NO Assay in culture medium of porcine CASMCs
The porcine CASMCs from second and third passage were seeded into 24-well cell culture
cluster at a density of 5 £ 105 cells/well in DMEM containing 10% FBS. After 24 h, the
culture medium was replaced with DMEM containing 2% FBS, and CASMCs were treated
with lipoprotein and drug, respectively. After another 24 h of culture, NO content in medium
was determined by NO test kit.
Effect of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside 427
3.7 Experiment grouping
3.7.1 Influence of THSG on lipoprotein oxidation induced by Cu21 or CASMCs. All
samples were treated with lipoprotein 60mg·mL21according to reference [1] and divided into
two main groups (LDL and VLDL groups). Then each main group was further randomly
divided into seven subgroups. Bland control groups: LDL (or VLDL); Positive control
groups: LDL (or VLDL) and Cu2þ of 10mmol·L21 (or CASMCs); Vit C groups: LDL
(or VLDL),Cu2þ of 10mmol·L21 (or CASMCs) and Vit C of 150mmol·L21; Four different
concentration THSG groups: THSG(0.1, 1, 10 or 100mmol·L21), LDL (or VLDL) and Cu2þ
of 10mmol·L21 (or CASMCs). The second to the third generation of cultured porcine
CASMCs were seeded into 24-well plates at a density of 2 £ 105 well and incubated at 378C
in DMEM without FBS. Lipoprotein (LDL, VLDL) and drugs were simultaneously added to
the cell, then incubated for another 24 h. MDA content of culture medium was evaluated by
the TBA method (using the MDA kit) and the oxidation level of lipoprotein was determined
by 1% agarose gel electrophoresis.
3.7.2 Influence of THSG on CASMCs proliferation and NO production induced by
LDL, VLDL, ox-LDL or ox-VLDL. The samples were divided into four (LDL, ox-LDL,
VLDL or ox-VLDL) main groups after adding CASMCs. Then, each main group was further
randomly divided into seven subgroups. Blank control groups: CASMCs; Positive control
groups: CASMCs and LDL (or VLDL, ox-LDL, ox-VLDL) of 60mg·mL21; Vit C groups:
CASMCs, LDL (or VLDL, ox-LDL, ox-VLDL) of 60mg·mL21 and Vit C of 150mmol·L21;
Four different concentration THSG groups: CASMCs, THSG (0.1, 1, 10 or 100mmol·L21)
and LDL (or VLDL, ox-LDL, ox- VLDL) of 60mg·mL21. The porcine CASMCs were
seeded into 96-well plates at a density of 5 £ 103 cells. After culturing for 24 h, the medium
was replaced to DMEM containing 2% FBS. LDL, ox-LDL, VLDL or ox-VLDL and drugs
were respectively added to each group. Each experimental group was added with drugs at
different concentration. Cultured for another 20 h, the proliferation of pig CASMCs was
measured by MTT assay.
3.8 Data analysis
Data are presented as mean ^ standard error (S.E.), and expressed as a percentage change
from the basal value for the unstimulated cells ( ¼ 100%). The paired student’s test was used
for statistical comparison. P , 0.05 was considered to be significant. P , 0.01 was
considered to be obviously significant.
References
[1] S. Koba, R. Pakala, T. Katagiri, C.R. Benedict. Atherosclerosis, 149, 61 (2000).[2] A.J. Lusis. Nature, 407, 233 (2000).[3] J. Seki, M. Nishio, Y. Kato, Y. Motoyama, K. Yoshida. Atherosclerosis, 117, 97 (2000).[4] G. Ryu, J.H. Ju, Y.J. Park, S.Y. Ryu, B.W. Choi, B.H. Lee. Arch. Pharm. Res., 25, 636 (2002).[5] J. Zou, Y. Huang, Q. Chen, E. Wei, K. Cao, J.M. Wu. Chin. Med. J. (Engl), 113, 99 (2000).[6] O. Araim, J. Ballantyne, A.L. Waterhouse, B.E. Sumpio. J. Vasc. Surg., 35, 1226 (2002).[7] J. Hwang, J. Wang, P. Morazzoni, H.N. Hodis, A. Sevanian. Free Radic. Biol. Med., 15 34, 1271 (2003).[8] P. Alaupovic, W.J. Mack, C. Knight-Gibson, H.N. Hodis. Arterioscler. Thromb. Vasc. Biol., 17, 715 (1997).[9] R.H. Boger, K. Sydow, J. Borlak, T. Thum, H. Lenzen, B. Schubert, et al. J. Circ. Res., 87, 99 (2000).
Q.-L. Liu et al.428
[10] S. Chattertee. Med. Cell. Biochem., 111, 143 (1992).[11] C.M. Shen, S.J. Mao, G.S. Huang, P.C. Yang, R.M. Chu. Life Sci., 1470, 443 (2001).[12] X. Yang, B. Cai, R.R. Sciacca, P.J. Cannon. Circ. Res., 74, 318 (1994).[13] Q.L. Liu, Y. Ban, J.H. Zhang, S.J. Ye, J.L. Wang. Chin. J. Pharmacol., (2004).[14] N. Auge, V. Garcia, F. Maupas-Schwalm, T. Levade, R. Salvayre, A. Negre-Salvayre. Arterioscler. Thromb.
Vasc. Biol., 22, 1990 (2002).[15] T.L. Cornwell, E. Arnold, N.T. Boerth, T.N. Lincoln. Am. J. Physiol. Cell. Physiol., 267, C1405 (1994).[16] A. Martin, B. Frei. Arterioscler. Thromb. Vasc. Biol., 17, 1583 (1997).[17] J.G. Zou, Y.Z. Huang, Q. Chen, E.H. Wei, T.C. Hsieh, J.M. Wu. Biochem. Mol. Biol. Int., 47, 1089 (1999).[18] Y. Ban, Q.L. Liu, Y. Jin, J.L. Wang. Chin. Tradi. Herb. Drug., 35, 1340 (2004).[19] R. Pakala, J.T. Willerson, C.R. Benedict. Circulation, 96, 2280 (1997).[20] C.B. Wang, Y.Q. Zong, W.S. Wu. Acta Univ. Med. Tongji., 24, 169 (1995).
Effect of 2,3,5,40-tetrahydroxystilbene-2-O-beta-D-glucoside 429
