Hindawi Publishing Corporation

Oxidative Medicine and Cellular Longevity

Volume 2012, Article ID 489690, 6 pages
doi:10.1155/2012/489690

Research Article
Cholesterol-Lowering Effect of Allicin on
Hypercholesterolemic ICR Mice

Yin Lu,1 Zhuojin He,2 Xiuying Shen,3 Xiaolu Xu,1 Jie Fan,4 Shaohua Wu,5 and Deyong Zhang1
1 College of Biological and Environmental Engineering, Zhejiang Shuren University, Hangzhou 310015, China
2 Instituteof Bioengineering, Zhejiang Academy of Medical Science, Hangzhou 310013, China
3 School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou 310023, China
4 Department of Biology, Shaoxing No.1 High School, Shengli West Road 1199, Yuecheng District, Shaoxing 312000, China
5 Cardiovascular Department, Integrated Chinese and Western Medicine Hospital, Hangzhou Shangcheng District,

Hangzhou 310008, China

Correspondence should be addressed to Yin Lu, [email protected]

Received 13 March 2012; Accepted 12 June 2012

Academic Editor: Michal Wozniak

Copyright © 2012 Yin Lu et al. This is an open access article distributed under the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Allicin was discussed as an active compound with regard to the beneficial effects of garlic in atherosclerosis. The aim of this study
was to investigate the cholesterol-lowering properties of allicin. In order to examine its effects on hypercholesterolemia in male
ICR mice, this compound with doses of 5, 10, or 20 mg/kg body weight was given orally daily for 12 weeks. Changes in body weight
and daily food intake were measured regularly during the experimental period. Final contents of serum cholesterol, triglyceride,
glucose, and hepatic cholesterol storage were determined. Following a 12-week experimental period, the body weights of allicin-
fed mice were less than those of control mice on a high-cholesterol diet by 38.24 ± 7.94% (P < 0.0001) with 5 mg/kg allicin,
39.28 ± 5.03% (P < 0.0001) with 10 mg/kg allicin, and 41.18 ± 5.00% (P < 0.0001) with 20 mg/kg allicin, respectively. A decrease
in daily food consumption was also noted in most of the treated animals. Meanwhile, allicin showed a favorable effect in reducing
blood cholesterol, triglycerides, and glucose levels and caused a significant decrease in lowering the hepatic cholesterol storage.
Accordingly, both in vivo and in vitro results demonstrated a potential value of allicin as a pronounced cholesterol-lowering
candidate, providing protection against the onset of atherosclerosis.

1. Introduction (RAS) are considered to be the key influencing factor of the

generation and development of AS [1, 2].
Atherosclerosis (AS) is one of the major risk factors in the Recently, various natural products have emerged as
development of hypertension and cardiovascular diseases. It active ingredients effective in controlling of AS [3, 4]. The
is the narrowing or occlusion of the arteries by plaque, which medicinal use of garlic (Allium sativum) has been known
consists of cholesterol, platelets, monocyte/macrophages, since the Ancient Egyptian era. For centuries, garlic was used
calcium, aggregating proteins, and other substances. Mor- for treating high blood pressure in China and Japan. Its
bidity of AS-induced coronary heart disease (CHD) gradu- reported beneficial effects include detoxification, antioxida-
ally elevates annually due to the improvement of life standard tion, antifungal, antibacterial activity, tumour suppression,
and the change of lifestyle in recent years. However, the and prevention of heart disease [5]. In relation to heart
mechanism of the onset and development of atherosclerotic disease, a major part of these publications deals with the
lesions are not completely understood until now. Many beneficial effect on the cardiovascular system, mainly related
complicated factors interaction and interrelated biological to AS. Garlic has been shown to alter blood lipids [6],
processes contribute to AS. Among these, high plasma levels decrease blood coagulability [7], and inhibit cell proliferation
of low-density lipoprotein (LDL), especially its oxidized form [8, 9]. Furthermore, garlic has been found to act as an anti-
(ox-LDL), and activation of the renin-angiotensin system hypertensive agent [10] and has been officially recognized
2 Oxidative Medicine and Cellular Longevity

as the treatment of hypertension by the Japanese Food and 60

Drug Administration [11]. Although the data available today 50

Body weight (g)

suggest that garlic contains biologically active compounds 40
which are beneficial in cardiovascular disease like AS, the
question about the active principles and their mechanism of 30
action is still not settled [5, 12]. 20
Allicin ((R,S)-diallyldisulfid-S-oxide), one of the sulfur 10
compounds from garlic, is formed by the action of the
0
enzyme alliinase on alliin. It possesses antioxidant activity 0 2 4 6 8 10 12
and is shown to cause a variety of actions potentially useful Time of test (week)
for human health [12]. Allicin exhibits hypolipidemic, anti-
Figure 1: Body weight changes (n = 6). Allicin was administered
platelet, and procirculatory effects. Moreover, it demon-
with doses (p.o.) of 0 (), 5 (), 10 (), and 20 () mg/kg,
strates antibacterial, anticancer, and chemopreventive activi-
respectively; the high cholesterol diet was fed throughout the test.
ties. The regular chow diet group (×) received no allicin treatment.
The background and the fact mentioned above focused The data are expressed as mean ± S.D. and are considered to be
our interest on the question of whether the known con- significantly different at P < 0.05 by the unpaired Student’s t-
stituent of processed garlic, allicin, which is discussed as test; P < 0.01 (from the 2th week of test) in 5 mg/kg allicin group;
an active component with regard to the beneficial effects of P < 0.0001 (from 1th week of test) in 10 and 20 mg/kg allicin
garlic preparation, has an effect on AS. Most previous studies groups.
used various garlic preparations in which allicin levels were
not well defined. In the present study, we investigated the
body weight, feed intake, and lipid profile in plasma, follow- 2.2. Biochemical Analysis of the Serum. Biochemical param-
ing hepatic cholesterol storage changes in the experimental eters in mouse plasma and lipoproteins at the end of
model of hypercholesterolemic ICR mice, to evaluate the study period were shown in Table 2. As shown in
whether application of a known amount of pure allicin has a the results, the high-cholesterol diet group obtained an
beneficial effect on formation of fatty streaks (AS) and blood elevated TC, TG, GLU, and LDL-C, but a decreased HDL-
lipid levels in mice. C, suggesting an effective induction of hypercholesterolemia
by supplementation of cholesterol in the diet, was effectively
2. Results established in ICR mice. The allicin administration in
doses of 5, 10, and 20 mg/kg lowered the elevated TC to
2.1. Changes in Body Weight and Feed Intake. The oral 75.94%, 56.92%, and 64.77% of high-cholesterol control,
administration of allicin to hypercholesterolemic ICR mice respectively. A similar decrease was seen in LDL-C level;
for 12 weeks resulted in a significant reduction in the body- the concentrations of which declined to 57.92%, 56.83%,
weight gain in a dose-dependent manner (Figure 1). The and 43.72% of control, respectively. The concentrations of
body weight increased from 19.70 ± 2.23 g, 20.07 ± 0.44 g, and HDL-C in all the allicin-treated animals, however, revealed
19.22±1.67 g at the start of the study to 31.93±3.01 g, 31.39± no significant differences except 5 mg/kg group. Table 2 also
2.42 g, and 30.41 ± 3.98 g at the termination in the groups showed that allicin administration lowered the elevated TG
administered with doses of 5, 10, and 20 mg/kg allicin, values to 63.03∼89.57% and GLU levels to 57.53∼62.00% of
respectively. Three test groups yielded reductions in body high-cholesterol control, respectively.
weight gain of 38.24 ± 7.94% (P < 0.0001), 39.28 ± 5.03%
(P < 0.0001), and 41.18 ± 5.00% (P < 0.0001), respectively, 2.3. Atherosclerotic Pathological Changes in the Liver. High
as compared to the high-cholesterol control. The high-cho- cholesterol diet stimulation could promote hyperlipidemia,
lesterol diet alone yielded no difference in body weight gain aggravated pathological changes of the liver, and even
when compared to the normal group (fed with a regular developed AS in the animals. Based on the results, the
chow diet), indicating that the supplementation of choles- morphology of hepatic cells in allicin-administered groups
terol itself had no appreciable effect on body weight gain. showed obvious pathological changes in a dose-dependent
A similar decline was seen in the daily food consumption manner compared with that of the high-cholesterol control
of allicin treatment (Table 1). In contrast to a previous report group (Figure 2). The lipid accumulation in hepatic cells in
[13], our observations demonstrated a significant decrease allicin administered groups became smaller and less than
in food consumption in a dose-dependent manner. This those of mice given by PBS as a placebo. However, there were
tendency was most pronounced in the first two weeks of no significant changes accompanied with fatty alteration and
the test. From that point on, it steadily regressed toward accretion of cells’ volume in the normal group, compared to
the control value until the end of the test period except in the mice at 5 weeks of age.
the 20 mg/kg allicin group, where the food intake remained
somewhat lower than in the control group throughout the 3. Discussion
experiment. At the 12th week of the test, the relative mean
feed intake was 82.0 ± 4.7%, 80.1 ± 4.3% and 67.7 ± 2.4%, In recent years, remarkable progress has been made in the
of the high cholesterol control in 5, 10, and 20 mg/kg allicin- prevention and treatment of AS. Atherosclerotic diseases
administered groups, respectively. such as ischemic heart disease, stroke, and peripheral arterial
Oxidative Medicine and Cellular Longevity 3

Table 1: Food intake efficiency during allicin administration (n = 6).

Week 1 2 3 11 12
Controld 100 ± 2.1 100 ± 3.7 100 ± 1.4 100 ± 4.9 100 ± 6.0
5 mg/kg allicine 90.3 ± 2.7a 88.1 ± 4.2 80.0 ± 2.3a 70.7 ± 5.9a 82.0 ± 4.7
10 mg/kg allicine 54.3 ± 0.5b 74.2 ± 11.7a 68.9 ± 1.6b 80.5 ± 3.1a 80.1 ± 4.3
20 mg/kg allicine 42.0 ± 1.3c 65.5 ± 5.7b 53.0 ± 3.3b 65.4 ± 3.2b 67.7 ± 2.4a
Normal (regular diet)e 89.9 ± 12.8a 85.6 ± 2.6a 82.3 ± 4.0a 93.4 ± 14.8 82.7 ± 15.9
The data are mean ± S.D. and significantly different at a P < 0.05, b P < 0.01, and c P < 0.001 by the unpaired Student’s t-test.
d The high cholesterol control group, 200 µL PBS (pH 7.4) was served instead of allicin as a negative control.
e The results are expressed as % of the high cholesterol control.

Table 2: Serum parameters after 12-week allicin administration (n = 6, mM).

5 mg/kg 10 mg/kg 20 mg/kg Controlc

Parameter Normal (regular diet)
High-cholesterol diet (allicin administered)
TC 43.91 ± 4.21b 32.91 ± 1.34b 37.45 ± 0.80b 57.82 ± 3.51b 32.99 ± 0.55
TG 1.66 ± 0.34a 1.33 ± 0.11b 1.89 ± 0.19b 2.11 ± 0.35b 1.04 ± 0.23
GLU 8.73 ± 0.49 8.22 ± 1.52 8.10 ± 0.14 14.08 ± 3.16b 8.38 ± 0.75
HDL-C 1.25 ± 0.20a 1.71 ± 0.59 1.60 ± 0.45 1.61 ± 0.16 1.94 ± 0.22
LDL-C 1.06 ± 0.52a 1.04 ± 0.22a 0.80 ± 0.15a 1.83 ± 0.50 1.57 ± 0.43
The data are mean ± S.D. and significantly different at a P < 0.05 and b P < 0.01 by the unpaired Student’s t-test.
TC: total cholesterol, TG: triglyceride, GLU: plasma glucose, HDL-C: high-density lipoprotein cholesterol, LDL-C: low-density lipoprotein cholesterol.
c The high cholesterol control group, 200 µL PBS (pH 7.4) was served instead of allicin as a negative control.

disease are associated with high serum cholesterol, male not be the best model for the proof of the current hypothesis.
gender, age, hypertension, cigarette smoking, diabetes, and Results here demonstrated that the daily administration of
so forth. Reduction in the concentration of blood lipids, pure allicin to ICR mice had a beneficial effect on the lipid
especially cholesterol, is a major goal in several primary and profile, causing a distinct decrease in the serum TC, LDL-C,
secondary prevention initiatives. Therefore, the approaches and accordingly a certain amount of decrease in the level of
to the prevention and treatment of atherosclerotic diseases TG and GLU. The reduction in LDL-C was more pronounced
are based primarily on the reduction of risk factors or rather than that of any other type of cholesterol. Since the
modifiable factors such as hyperlipidemia, hypertension, concentration of HDL-C was not significantly changed, the
and diabetes. This approach can be regarded as indirect cholesterol-lowering effect was in fact exclusively attributed
antiatherosclerotic therapy. to the decline of LDL-C, making the most characteris-
Several studies have suggested that garlic may have bene- tic changes involved in cholesterol modulation by allicin.
ficial effects on plasma cholesterol levels [14–16], while other Similar results that allicin exerted a beneficial effect on the
studies found no influence [15–18]. We suggest that the com- lipid profile in hyperlipidemic rabbits had been previously
position and quantity of sulfur components of different pro- reported [20].
tocol designs of garlic preparations used in various studies On the other hand, the results also showed a significant
and the different mouse models used could account in part decrease in lowering the hepatic cholesterol storage as com-
for the inconsistent findings. The pharmacological activities pared with the controls. The lipid accumulation in hepatic
of garlic are attributed to the thiosulfinate compounds, of cells has decreased as well, suggesting that allicin alleviated
which allicin is approximately 75% [17]. Therefore, garlic liver stress related to hypercholesterolemia and thus was
antiatherosclerotic properties are mainly attributed to allicin. advantageous to prevent from becoming a fatty liver.
The use of pure allicin to study the atheroprotective effect of Moreover, food consumption in allicin-treated animals
garlic is reasonable. showed a remarkable decline in a dose-dependent manner.
In the present study, by using pure allicin, we could Food consumption can play a role in cholesterol homeostasis
investigate the effect of a well-defined component of garlic on as an independent influential factor. Since the restriction in
AS and to seek for possible mechanisms of its antiatherogenic food intake is also correlative to blood cholesterol reduction,
activity. Authors of many studies have shown the relationship it is quite possible that allicin has affected cholesterol home-
between high-cholesterol levels, especially HDL-C and LDL- ostasis by a simple but efficient way of food intake suppres-
C, and the development of AS [18]. The high-cholesterol sion, as well as the change of body weight. And the actions
diet could induce a rise in blood cholesterol level to more of the cholesterol lowering presented in this paper would
physiologic levels, for which the rise became unphysiologic endow more nutritional and therapeutic value with allicin.
[19]. We chose not to use the transgenic mouse models of In summary, the present study confirmed and extended
AS, although they resemble human AS better, because these the understanding that allicin may beneficially affect the
models have a very strong phenotype and therefore might risk factors for AS—hyperlipidemia and attribute to lower
4 Oxidative Medicine and Cellular Longevity

(a) (b) (c)

(d) (e) (f)

Figure 2: Photomicrographs of the section surface of livers stained with Oil Red O. (a) ICR mice (5 weeks age), fed with high cholesterol
diet for one week before test; (b) normal group (17 weeks age), fed with a regular chow diet for 12 weeks; (c) high cholesterol control group
(17 weeks age), fed with high cholesterol diet plus PBS for 12 weeks; (d) 5 mg/kg allicin administered group (17 weeks age), fed with high
cholesterol diet for 12 weeks; (e) 10 mg/kg allicin administered group (17 weeks age), fed with high cholesterol diet for 12 weeks; (f) 20 mg/kg
allicin administered group (17 weeks age), fed with high cholesterol diet for 12 weeks. Magnification: ×400.

the hepatic cholesterol storage. Thus, the active component To minimize oxidation, the diets were stored in the dark at
allicin could potentially provide protection against the onset 4◦ C until use. The animals were given ad libitum with the
of AS. However, it is noteworthy that allicin is very reactive regular chow diet and tap water for one week prior to any
and rapidly converted to its metabolites [21], which may experiment. Then after the mice had been fed with high-
limit its biological activity. Further studies to elucidate the cholesterol diet for one week, only those with over 200 mg/dL
exact mechanism and the molecular basis of action of allicin of blood cholesterol were selected for use in the experiment.
on various animal species and in particular on humans are Some sources recommend that allicin should be used in
required. high doses for medicinal purposes [13, 22]. In our prelim-
inary study, three means of administration, intraperitoneal
4. Subjects and Methods (ip), intravenous (iv) and oral (po) were tested for allicin. For
oral administration for the dose-range experiment, higher
4.1. Preparation of Allicin. Allicin, synthesized compound concentrations of allicin-administered op (25 mg/kg/day)
with purity more than 95%, was obtained from Lichtwer caused discomfort to the animals and were discontinued.
Pharma GmbH (Germany) and dissolved in phosphate- And a regimen of 2 mg/kg/day was ineffective. We therefore
buffered saline (PBS). The solution was stored in dark at 4◦ C tested a modified version of the allicin po regimen. Twenty-
until use (not more than 3 months). The chemical stability of four selected mice were divided into four groups (n = 6)
allicin was assessed using HPLC and quantitatively deter- by randomization, three allicin test groups and one high-
mined at particular time intervals. cholesterol control group. To the three test groups, 200 µL
allicin solution in PBS was orally administered to correspond
4.2. Oral Administration of Allicin to Hypercholesterolemic to 5, 10, or 20 mg allicin/kg body weight once per day for a
ICR Mice. Male ICR mice (3 weeks, 10 ± 2 g) were provided period of 12 weeks (the amount of allicin administered had
by the Animal Center, Academy of Chinese Traditional been found to be safe in a preliminary study). To the high-
Medicine, Zhejiang, China. All mice were raised in a 12/12- cholesterol control group, 200 µL PBS (pH 7.4) was served
hour light-dark cycle room with controlled temperature (21– instead of allicin as a negative control. All the above animals
23◦ C) and humidity (50–60%). The mice were fed two kinds were provided with the high-cholesterol diet throughout the
of diet. The atherogenic high-cholesterol diet contained animal test. A normal group (n = 6) fed with a regular chow
15.75% fat (43% saturated fat) and 1.25% cholesterol diet was used for comparison and was supplied with the same
(TD88137; Harlan). The regular chow diet consisted of volume of PBS (pH 7.4) instead of allicin. The mice were
4.5% fat by weight (0.02% cholesterol) (TD19519; Koffolk). observed at least twice a day for clinical abnormalities.
Oxidative Medicine and Cellular Longevity 5

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