N U TR IT ION RE S EA RCH 3 2 ( 2 0 12 ) 71 8 7 26

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Panax ginseng reduces oxidative stress and restores

antioxidant capacity in aged rats

Thiyagarajan Ramesh a , Sung-Won Kim a , Seock-Yeon Hwang b , Sang-Hyun Sohn a ,

Sung-Kwang Yoo a , Si-Kwan Kim a,
a
Department of Life Science, College of Biomedical and Health Science, Institute of Biomedical and Health Science, Konkuk University,
Chungju-si 380-701, Korea
b
Department of Biomedical Laboratory Science College of Applied Science and Industry, Daejeon University, Daejeon-si, Korea

A R T I C LE I N FO AB S T R A C T

Article history: Nutritional antioxidants interact with cells in an active mode, including retrieving and sparing
Received 7 May 2012 one another, to diminish oxidative stress. However, the intracellular balance of prooxidants
Revised 24 June 2012 and antioxidants becomes unbalanced, favoring prooxidants during the aging process. One
Accepted 7 August 2012 hypothesis is that an aging-associated increase in oxidative stress is the primary cause of
aging. Hence, the research hypothesis for this study is that Korean red ginseng reduces
Keywords: oxidative stress in vivo. Therefore, we investigated the efficacy of Korean red ginseng water
Antioxidants extract (GWE) in reducing aging-associated oxidative stress by measuring lipid peroxidation
Panax ginseng nutrients and antioxidant levels in older rats compared with young rats. We observed a significant
Aging increase in the markers for oxidative damage (eg, lipid peroxidation) and markers for vital
Oxidative stress organ damage (eg, aspartate aminotransferase, alanine aminotransferase, urea, and
Lipid peroxidation creatinine levels) in aged rats. The oxidative damage was accompanied by a significant
Rats decrease in enzymatic antioxidants such as superoxide dismutase, catalase, glutathione
peroxidase, glutathione reductase, and glutathione-S-transferase, and nonenzymatic
antioxidants such as reduced glutathione, vitamin E, and vitamin C. Aged rats fed a diet
supplemented with Korean red ginseng water extract had significantly less oxidative damage,
possibly by enhancing the enzymatic and nonenzymatic antioxidants status. Our data
suggest that consumption of Korean red ginseng reduces lipid peroxidation and restores
antioxidant capacity by suppressing oxidative stress in rats.
2012 Elsevier Inc. All rights reserved.

1. Introduction been used as a tonic in traditional Oriental medicine for

more than 2000 years; it is well known as an energy booster
Panax ginseng (P ginseng), one of the best health foods for and dietary supplement in Korea, China, Japan, and other
vitality and combating fatigue, increases energy and elimi- Asian countries. For example, Korean red ginseng (KRG), in
nates chronic fatigue while improving health. Ginseng has particular, is well documented as a natural health food. It is

Abbreviations: ALT, alanine aminotransferase; AST, aspartate aminotransferase; BUN, blood urea nitrogen; CAT, catalase; CDNB, 1-
chloro-2,4-dinitro benzene; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GST, glutathione-S-
transferase; GWE, Korean red ginseng water extract; KRG, Korean red ginseng; MDA, malondialdehyde; ROS, reactive oxygen species; SOD,
superoxide dismutase.
Corresponding author. Tel.: +82 43 480 3574; fax: +82 43 840 3872.
E-mail address: skkim@kku.ac.kr (S.-K. Kim).

0271-5317/$ see front matter 2012 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.nutres.2012.08.005
 N U TR IT ION RE S E ARCH 3 2 ( 2 0 12 ) 71 8 7 2 6
well known that emperor Shi Huang Di (founder of the Qin
dynasty, 221-206 BC), the first Chinese sovereign to proclaim
himself emperor, had eagerly sought antiaging herbs, and
attempts to counteract aging have been pursued since
ancient times. Panax ginseng is one of the representative
botanicals used empirically to combat age-related organ
dysfunction [1,2].
Ginsenosides (ginseng saponins) are the major active
components of ginseng and are used as markers for deter-
mining the quality of ginseng [3,4]. Korean red ginseng has
been developed for long-term storage and distribution. Red
ginseng is produced by steaming and drying fresh ginseng.
During this process, ginsenosides undergo chemical changes
that have the potential to bring about special physiologic
activities in vivo [5]. Currently, more than 30 different
ginsenosides from KRG have been isolated and characterized,
and these ginsenosides are known to have different pharma-
cologic effects. Based on their aglycone moieties, ginsenosides
can be classified into 2 major categories: 20(S)-protopanax-
adiol (ginsenosides Rb1, Rb2, Rc, Rd, Rg3, and Rs3) group and
20(S)-protopanaxatriol (ginsenosides Re, Rg1, and Rf) group [6].
Several investigators have discovered new ginsenosides
from red ginseng, which are not usually found in raw ginseng.
These ginsenosides are Rg5, Rg6, Rh2, Rh4, and Rg3 [7,8]. Other
than the ginsenosides, several other compounds are present
in the roots of the ginseng plant. Twelve kinds of phenolic
compounds including salicylic acid, caffeic acid, and maltol
have been isolated from ginseng. Maltol, which is present only
in red ginseng and is produced from maltose by amino-
carbonyl reaction, shows antioxidant activity [7]. In addition,
the ginseng root contains amino acids; vitamins A, B1, B2, B12,
C, and E and niacin; and inorganic elements such as sodium,
potassium, calcium, magnesium, phosphorus, iodine, iron,
zinc, copper, manganese, and selenium [911]. The impor-
tance of these and other micronutrients is believed to be
essential for living organisms.
According to the free radical theory of aging [12], the
generation of reactive oxygen species (ROS) or free radicals
can lead to cell and tissue damage, along with changes in
cellular function resulting in aging and early cell death.
Several natural defense mechanisms protect against the
toxic effects of free radicals. Enzymatic antioxidants such as
superoxide dismutase (SOD; which eradicates superoxides in
the presence of copper and zinc), glutathione peroxidase
(GPx; which converts hydrogen peroxide into water and
converts various hydroperoxides into less harmful hydrox-
ides in the presence of selenium), catalase (CAT; which can
also break down hydrogen peroxide as well as nonenzymatic
antioxidants such as vitamins C and E), and reduced
glutathione (GSH) scavenge ROS and free radicals or prevent
their formation.
The term oxidative stress depicts a state of imbalance
between the generation of ROS and antioxidant defenses,
either induced by an endogenously or exogenously derived
elevation in ROS production or by a reduction in defense
mechanisms resulting in cell and tissue damage. Several
studies have shown that oxidative stress causes structural
and functional alterations in molecular, cellular, tissue, and
organ systems [13,14]. Antioxidant nutrients play a vital role
in the body's defense against elevated levels of free radicals
719
and delay the onset of aging and age-associated degenerative
diseases [1517]. In this respect, food that contains antioxi-
dant properties may be effective and useful for reducing the
risk of aging-associated oxidative damage. Taken together,
these lines of evidence suggest that KRG may be an effective
antioxidant nutrient against oxidative stressinduced alter-
ations due to aging. Until now, research has tended to focus on
the therapeutic value of KRG; however, there is relatively little
information pertaining to its antioxidant activity and its
possible use for attenuating oxidative stress during aging.
Therefore, the aim of this study is to test the hypothesis that
KRG reduces oxidative stress in vivo. The research objectives
are to demonstrate oxidative stress in aged rats and to
determine whether a water extract of the KRG increases
antioxidant levels in older rats.
2. Methods and materials
2.1. Preparation of KRG-water extract
Korean red ginseng water extract (GWE) was kindly supplied
by Korea Ginseng Corporation (Seoul, Korea). Red ginseng was
prepared by steaming and sun drying and then extracted with
10 volumes of tap water at 85C for 48 hours and was filtered.
The filtrate was concentrated under reduced pressure to
obtain GWE resulting in the dark-brown syrup.
2.2. Analysis of ginsenosides
The GWE was dissolved in ultrapure distilled water and
passed through a SepPak C18 cartridge (Waters Corporation,
USA); the cartridge was washed with 20% methanol, and
the saponin fraction was eluted with absolute methanol for
the analysis of ginsenosides, as described in European
Pharmacopoeia [18]. The content of crude saponin in KRG-
water extract is approximately 7%, and it is composed of
the following ginsenosides: 8.27 mg/g of Rb1, 3.22 mg/g of
Rb2, 3.90 mg/g of Rc, 1.09 mg/g of Rd, 2.58 mg/g of Re, 1.61
mg/g of Rf, 2.01 mg/g of Rg1, 1.35 mg/g for (20S)-Rg2, 1.04
mg/g for (20S)-Rg3, and 0.95 of Rh1, respectively.
2.3. Animals and diets
Male Sprague-Dawley rats (20 rats were 12 months old [750
20 g] and 10 rats were 2 months old [280 10 g]) were
purchased from Samtako Bio Korea (Osan, Korea) and
acclimatized to the animal facility for at least 1 week before
the experiment. They were provided with a standard pelleted
diet (the ingredient composition of the diets is shown in
Table 1) and water ad libitum and housed at constant
temperature (23 2C) and relative humidity (55% 10%) on
a 12-hour light/dark cycle. Food consumption was measured
daily, and body weight was recorded weekly. The rats were
housed in the Regional Innovation Center experimental
animal facility at Konkuk University in accordance to the
institutional animal care and use committee guidelines. The
study protocol and use of rats were approved by the animal
ethics committee (ethical no. ku08080) in accordance to the
14th article of Korean animal protection law.
720 N U TR IT ION RE S EA RCH 3 2 ( 2 0 12 ) 71 8 7 26

Table 1 Ingredient composition of the experimental 2.6. Assessment of lipid peroxidation marker
diets fed to rats
Amount (g) Lipid peroxidation was determined by the procedure of
Ohkawa et al [19]. One of the major secondary by-products
Ingredient or nutrient content
of lipid peroxidation is malondialdehyde (MDA). Malondialde-
Casein (from milk) 140
Corn starch 466 hyde, along with other by-products, reacts with thiobarbituric
Sucrose 100 acid to generate a colored product with maximum absorption
Dextrose 155 at 532 nm, representing the color produced by all the
Cellulose 50 thiobarbituric acid reactive substances. Malondialdehyde
Soybean oil 40 was expressed as nanomoles per milligram of protein
AIN-93 mineral mix a 35
throughout the text.
AIN-93 vitamin mix a 10
t-Butyl hydroquinone 0.008
L-Cystine 1.8 2.7. Analysis of on enzymatic antioxidants
Choline bitartrate 2.5
Total (g/kg) 1000 Copper-zinc SOD (Cu-Zn SOD) was assayed by the method of
Protein (kcal, %) 15 Marklund and Marklund [20]. The degree of inhibition of the
Carbohydrate (kcal, %) 77
auto-oxidation of pyrogallol at an alkaline pH by Cu-Zn SOD
Fat (kcal, %) 8
was used as a measure of the enzyme activity. The unit of
Energy (kcal/kg) 3850
enzyme activity is expressed as 50% inhibition of auto-
a
Mineral and vitamin mixture for AIN-93 rodent diet. This diet was oxidation of pyrogallol per minute per milligram of protein.
provided to young and aged control rats. Korean red ginseng water The activity of CAT was measured by the method of Sinha [21].
extract was mixed into the diet and pelletized and then given to
The principle of this method is that dichromate in acetic acid is
aged rats (designated as GWE administered rats). Korean red
ginseng water extract content was adjusted by weighing the aged
reduced to chromic acid when heated in the presence of H2O2,
rats weekly and the daily dietary intake. Korean red ginseng water with the formation of perchloric acid as an unstable interme-
extract was administered for 120 days to aged rats. diate; the chromic acetate thus produced is measured at 570
nm. The unit of enzyme activity is expressed as micromoles of
H2O2 consumed per minute per milligram of protein.
The activity of selenium-dependent GPx (Se-GPx) was
2.4. Experimental design and administration of GWE determined by the method of Rotruck et al [22] using hydrogen
peroxide as a substrate in the presence of GSH. The unit of
The rats were assigned to the following 3 groups: (1) young enzyme activity is expressed as micromoles of GSH consumed
rat control group (n = 10), (2) aged rat control group (n = 10) per minute per milligram of protein. Glutathione reductase
receiving vehicle only, and (3) aged rats treated with GWE (GR), uses NADPH (nicotinamide adenine dinucleotide phos-
(n = 10) at a daily dosage of 200 mg/kg body weight for 4 phate hydrogen) to convert oxidized glutathione to GSH, was
months. The GWE was mixed evenly with sterilized assayed by the method of Mize and Langdon [23]. The unit of
standard diet powder and administered orally after pellet- enzyme activity is expressed as micromoles of NADPH
ing. The appropriate amount of GWE was adjusted by oxidized per minute per milligram of protein. Glutathione-S-
weighing the rats weekly and also their daily dietary intake. transferase (GST) activity was analyzed by the method of
At the end of the experimental period, all animals were Habig et al [24]. The conjugation of glutathione to 1-chloro-2,4-
fasted for 24 hours and euthanized under general anesthe- dinitro benzene (CDNB) was measured as a nonspecific
sia with diethyl ether. Blood was collected directly from the substrate for GST activity. The unit of enzyme activity is
heart, and serum was separated by centrifugation for expressed as micromoles of CDNB used per minute per
enzyme analysis. The liver, kidney, heart, and lung were milligram of protein.
excised and washed in ice-cold saline solution, and the
adhering fat and connective tissues were removed. A 10% 2.8. Estimation of nonenzymatic antioxidants
homogenate of the tissue was prepared in Tris-HCl buffer
(0.1 M; pH 7.4) and centrifuged (2500 rpm for 10 minutes at Reduced glutathione was measured by the method of Beutler
4C) to pellet the cell debris, and the supernatant was et al [25]. 5,5-Dithio (2-nitrobenzoic acid) is a disulfide
harvested for biochemical assays. compound that is readily reduced by sulfhydryl compounds,
forming a brightly colored yellow anion; the optical density of
2.5. Determination of serum markers this yellow substance is measured at 412 nm. Reduced
glutathione concentration was expressed as micrograms per
To measure biochemical parameters, blood was collected in milligram of protein. Vitamin C was measured by the method
an SST gel and clot activator tube (Becton Dickinson, Franklin of Omaye et al [26]. Vitamin C is oxidized by copper to form
Lakes, NJ, USA). Serum was separated by centrifugation (1500 dehydroascorbic acid, which reacts with 2,4-dinitrophenyl
g, 10 minutes, room temperature). An automated chemistry hydrazine to form bis-2,4-dinitrophenyl hydrazine and un-
analyzer (Hitachi-747; Hitachi Medical, Tokyo, Japan) was used dergoes further rearrangements to form a product with an
to measure serum levels of aspartate aminotransferase (AST), absorption maximum at 520 nm. Thiourea provides a reducing
alanine aminotransferase (ALT), blood urea nitrogen (BUN), medium that helps to prevent interference from nonascorbic
and serum creatinine. acid chromogens. The concentration of vitamin C is expressed
 N U TR IT ION RE S E ARCH 3 2 ( 2 0 12 ) 71 8 7 2 6 721

Table 2 Effect of GWE on body weight and food intake of

aged rats 3. Results
Aged Aged + GWE
3.1. Effect of GWE on body weight and food intake
Initial (g) 752.00 25.07 748.17 12.73NS
Body weight final (g) 809.00 14.13 804.50 17.14NS Change in body weight and food intake after 120 days of
Gain (g/d) 57.00 26.62 56.33 18.58NS
administration of GWE to aged rats is presented in Table 2.
Food intake (g/d) 15.10 1.94 14.21 2.52NS
Supplementation with GWE did not result in a significant
Values are presented as means SDs (n = 10). Korean red ginseng change in body weight gain of the aged rates compared with
water extract was administered for 120 days. Statistical comparison those not given GWE. No significant differences were observed
was made between aged rats and aged + GWE. P < .05 was
in food intake between the 2 groups.
considered significant. NS, nonsignificant, using the Tukey method.

3.2. Effect of GWE on lipid peroxidation

as milligrams per gram of wet tissue. The amount of vitamin E
content was determined by the procedure of Varley et al [27].
The content of vitamin E is expressed as micrograms per gram
of wet tissue.
2.9. Statistical analyses
The data obtained were subjected to 1-way analysis of
variance, and a post hoc test was performed for intergroup
comparisons using Tukey multiple comparison with the
GraphPad prism software package (version 3.0; GraphPad
Software. Inc., CA, USA) for Windows. Values are expressed
as the means SDs for 10 animals in each group. At P < .05,
differences were considered significant.
Lipid peroxidation was measured by the formation of MDA in
the homogenates of the liver, kidneys, heart, and lungs from
young rats and aged rats supplemented with or without GWE
(Fig. 1). Aged rats had significantly higher (P < .001) MDA
levels compared with the young control rats. Treatment for
aged rats with GWE lowered the MDA (P < .001) concentration
such that it was nearly the same as normal young rats,
presumably by limiting lipid peroxidation in the tissues of
the liver, kidneys, heart, and lungs.
3.3. Effect of GWE on serum biomarkers
Activities of AST and ALT were significantly increased (P <
.001) in the serum of aged rats not supplemented with GWE

Fig. 1 Effect of GWE on lipid peroxidation in the liver (A), kidneys (B), heart (C), and lungs (D) of aged rats. Values are presented
as the means SDs (n = 10). aAged rats compared with young rats. bAged + GWE compared with aged rats. *Significance at P <
.001, using the Tukey method.
722 N U TR IT ION RE S EA RCH 3 2 ( 2 0 12 ) 71 8 7 26

Fig. 2 Effect of GWE on serum AST (A), ALT (B), BUN (C), and creatinine (D) levels in aged rats. Values are presented as the
means SDs (n = 10). aAged rats compared with young rats. bAged + GWE compared with aged rats. *Significance at P < .001,
using the Tukey method.

compared with the values in young rats (Fig. 2). Adminis-
tration of GWE to aged rats showed significant decreases (P <
.001) in the activities of these enzymes when compared with
the untreated aged rats (Fig. 2). Similarly, BUN and serum
creatinine concentrations were significantly increased (P <
.001) in aged rats compared with the young rats (Fig. 2).
Korean red ginseng water extract administration to the aged
rats significantly lowered (P < .001) the levels of BUN and
creatinine when compared with the untreated aged rats
(Fig. 2).
3.4. Effect of GWE on enzymatic antioxidants
The activities of Cu-Zn SOD and CAT enzymes were signifi-
cantly decreased (P < .001) in the liver, kidneys, heart, and
lungs of aged untreated rats when compared with the values
in young rats (Tables 3 and 4). Likewise, activities of GSH-
dependent enzymes such as GPx, GR, and GST were signifi-
cantly reduced (P < .001) in the liver, kidneys, heart, and lungs
of aged untreated rats compared with those in young rats
(Tables 3 and 4). In aged rats administered GWE, the activities

Table 3 Effect of GWE on enzymatic antioxidants in the liver and kidney of young and aged rats
Liver Kidney

Young rats Aged rats Aged + GWE Young rats Aged rats Aged + GWE
a b a
SOD 16.19 1.45 8.87 1.2 14.60 1.32 14.29 1.83 8.61 1.34 15.31 0.88 b
CAT 171.47 21.26 68.35 5.67 a 143.16 29.04 b 352.38 29.69 242.71 29.63 a 347.64 15.52 b
GPx 6.60 1.13 2.83 0.23 a 6.27 1.16 b 20.21 4.69 7.84 0.89 a 18.12 1.76 b
GR 6.74 0.98 3.40 0.38 a 6.44 1.38 b 7.35 1.81 3.53 0.90 a 7.17 1.47 b
GST 4.75 0.71 1.85 0.24 a 4.88 0.93 b 1.59 0.24 0.82 0.15 a 1.56 0.34 b

Units: SOD, units per minute per milligram of protein (1 U is equal to the amount of enzyme that inhibits the pyrogallol auto-oxidation by 50%);
CAT, micromoles of H2O2 consumed per minute per milligram of protein; GPx, micromoles of GSH consumed per minute per milligram of
protein; GR, micromoles of NADPH oxidized per minute per milligram of protein; GST, micromoles of CDNB used per minte per milligram of
protein. Korean red ginseng water extract was administered for 120 days. Values are presented as means SDs (n = 10).
a
Aged rats compared with young rats.
b
Aged + GWE compared with aged rats.
Significance at P < .001, using the Tukey method.
Significance at P < .01, using the Tukey method.
 N U TR IT ION RE S E ARCH 3 2 ( 2 0 12 ) 71 8 7 2 6 723

Table 4 Effect of GWE on enzymatic antioxidants in heart and lung of young and aged rats
Heart Lung

Young rats Aged rats Aged + GWE Young rats Aged rats Aged + GWE
a b a
SOD 15.07 1.10 7.23 0.85 11.83 1.53 13.40 1.54 9.44 0.79 14.05 2.29 b
CAT 145.16 17.60 73.94 22.39 a 138.12 19.29 b 131.83 6.54 65.06 25.81 a 119.79 4.33 b
GPx 51.26 7.86 26.73 4.12 a 48.01 7.93 b 32.50 3.67 14.26 2.99 a 28.91 7.78 b
GR 8.20 0.81 2.19 0.59 a 6.73 0.79 b 8.40 1.69 2.23 1.20 a 6.89 1.61 b
GST 1.68 0.31 0.59 0.19 a 1.72 0.22 b 1.83 0.14 0.71 0.14 a 1.82 0.11 b

Units: SOD, units per minute per milligram of protein (1 U is equal to the amount of enzyme that inhibits the pyrogallol auto-oxidation by 50%);
CAT, micromoles of H2O2 consumed per minute per milligram of protein; GPx, micromoles of GSH consumed per minute per milligram of
protein; GR, micromoles of NADPH oxidized per minute per milligram of protein; GST, micromoles of CDNB used per minte per milligram of
protein. Korean red ginseng water extract was administered for 120 days. Values are presented as means SDs (n =10).
a
Aged rats compared with young rats.
b
Aged + GWE compared with aged rats.
Significance at P < .001, using the Tukey method.
Significance at P < .01, using the Tukey method.

of these enzymes reverted to near-normal levels; that is, there
were no significant differences compared with the values
observed in young rats (Tables 3 and 4).
3.5. Effect of GWE on nonenzymatic antioxidants
A significant decrease in the levels of GSH was observed (P <
.001) in the liver, kidneys, heart, and lungs of aged untreated
rats when compared with the levels in those given GSH and in
normal young rats (Tables 5 and 6). Administration of the GWE
to aged rats showed increased levels of GSH (P < .001) that
approached the levels found in young rats (Tables 5 and 6).
Similarly, a significant decrease in the levels of vitamins C and
E (P < .001) were observed in the liver, kidneys, heart, and lungs
of aged untreated rats compared with normal young rats
(Tables 5 and 6). However, the levels of vitamins C and E in the
liver, kidneys, heart, and lungs of aged rats administered GWE
were found to be near the levels observed in young rats
(Tables 5 and 6).
4. Discussion
Food and beverages contain a wide variety of nutrients and
substances that potentially improve health and may reduce
the effects of aging. Some of the nonnutrients in food have
been called phytochemcials. Some phytochemicals have anti-
oxidant activity. During the aging process, our body becomes
gradually more susceptible to the long-term effects of
oxidative stress and inflammation at the cellular level. The
theory is that antioxidants and other age-defying compounds
help cells to ward off damage from free radicals and minimize
the impact of aging. In this study, P ginseng was found to have
antioxidant and antiaging properties in aged rats. Lipid
peroxidation is known to be a deteriorative cellular reaction
that contributes to aging [28]. Because free radical species
derived from molecular oxygen are highly charged, their
interaction with lipids results in oxidation and formation of
lipid peroxides. We found that the level of MDA, which is an
indicator of lipid peroxidation, was increased in the liver,
kidneys, heart, and lungs of aged rats compared with young
rats. This might be due to excessive free radicals and
insufficient levels of antioxidants needed to scavenge peroxyl
radicals generated during aging [29]. This observation is
supported by data from several studies that show significant
increases in the levels of MDA in aged rats [15,3034]. Korean
red ginseng water extract treatment for aged rats resulted in a
significant decrease in the levels of MDA in the liver, kidneys,
heart, and lungs. This is consistent with a recent report
showing that KRG extract reduces MDA levels in aflatoxin B1

Table 5 Effect of GWE on nonenzymatic antioxidants in liver and kidney of young and aged rats
Liver Kidney

Young rats Aged rats Aged + GWE Young rats Aged rats Aged +GWE
a b a
GSH 6.68 1.55 2.91 0.32 6.35 1.32 1.56 0.31 0.69 0.13 1.55 0.35 b
Vitamin C 0.55 0.04 0.24 0.03 a 0.47 0.07 b 0.32 0.01 0.14 0.01 a 0.28 0.02 b
Vitamin E 24.75 3.58 15.08 1.56 a 22.50 3.16 b 4.79 0.09 3.13 0.95 a 4.80 0.10 b

GSH expressed as micrograms per milligram of protein. Vitamin C expressed as milligrams per gram of wet tissue. Vitamin E expressed as
micrograms per gram of wet tissue. Korean red ginseng water extract was administered for 120 days. Values are presented as means SDs (n = 10).
a
Aged rats compared with young rats.
b
Aged + GWE compared with aged rats.
Significance at P < .001, using the Tukey method.
Significance at P < .01, using the Tukey method.
724 N U TR IT ION RE S EA RCH 3 2 ( 2 0 12 ) 71 8 7 26
Table 6 Effect of GWE on nonenzymatic antioxidants in heart and lung of young and aged rats
Heart
Young rats Aged rats Aged + GWE
a b
GSH 3.69 0.46 1.17 0.43 3.33 0.60
Vitamin C 0.61 0.13 0.24 0.07 a 0.57 0.14 b
Vitamin E 15.40 0.51 7.76 1.20 a 14.02 0.38 b
GSH expressed as micrograms per milligram of protein. Vitamin C expressed as milligrams per gram of wet tissue. Vitamin E expressed as
micrograms per gram of wet tissue. Korean red ginseng water extract was administered for 120 days. Values are presented as means SDs (n = 10).
a
Aged rats compared with young rats.
b
Aged + GWE compared with aged rats.
Significance at P < .001, using the Tukey method.
treated rats [35]. These results suggest that GWE can reduce
aging-related free radicals and help to maintain normal cell
function. This effect is presumably due to the many antiox-
idants in P ginseng including ginsenosides, vitamins, sulfur-
containing amino acids, trace elements, and other active
compounds [611].
Damage to organs in vivo caused by free radicals is
evidenced by the elevation of biomarkers in serum of aged
rats [34,36,37]. These markers are used to evaluate organ
dysfunction and are indicative of cellular damage, cell leakage,
and the loss of cell membrane integrity in the liver, kidney,
heart, and other organs. We observed a significant elevation in
serum AST and ALT activities, and BUN and creatinine
concentrations in aged rats. The increased expression of
these markers is consistent with ROS-induced oxidative
damage in these organs. In this study, we also report that
administration of GWE protected the organs from oxidative
damage as evidenced by decreased AST and ALT activities and
lower BUN and creatinine levels in aged rats. Therefore, GWE
has potential to reduce free radicals, and our results are
consistent with the observation that KRG extract reduces AST
and ALT activities in aflatoxin B1treated rats [35].
We found that Cu-Zn SOD activity was significantly lower
in the liver, kidneys, heart, and lungs of aged rats. This might
be due to ROS-induced enzyme degradation or reduced
synthesis of the SOD as age advances; however, the decline
in SOD activity indicates inefficient scavenging of superoxide
radicals [15,3034]. On administration of GWE, SOD activity
was restored to near-normal levels in the liver, kidneys, heart,
and lungs of aged rats compared with young rats. This might
be due to the effects of the nutrients in ginseng including
ginsenosides, antioxidant vitamins, sulfur-containing amino
acids, trace elements, and other active compounds [611].
Copper is essential for Cu-Zn SOD catalytic activity as a
facilitator of electron transfer. Zinc also plays an important
role in maintaining the structure and function of Cu-Zn SOD
[38]. Some studies have reported that Cu-Zn SOD activity is
significantly decreased because of oxidative stress and Zn
deficiency in rats [39]. The low enzyme activity might be due
to aging-related oxidative stress and decreased dietary copper
intake as a consequence of diminished food consumption
caused by Zn deficiency [40]. Copper is an essential nutrient in
the human diet because it is involved in the proper use of iron
and, especially, for the synthesis of important biomolecules
including Cu-Zn SOD. Because, P ginseng contains trace
Lung
Young rats Aged rats Aged + GWE
a
10.63 1.79 4.84 0.78 9.10 0.91 b
0.62 0.16 0.25 0.08 a 0.58 0.11 b
14.32 0.28 8.10 0.97 a 14.53 0.74 b
elements such as Cu and Zn [911], these cofactors might
enhance the activity of Cu-Zn SOD in aged rats.
Catalase activity was significantly decreased in the liver,
kidneys, heart, and lungs of aged rats, and this can be
attributed to increased production of free radicals in older
animals [15,3134]. The reduction in CAT activity might be due
to ineffective scavenging of H2O2resulting in elevated H2O2
levels, which can react with O2 to give OH radicals and
increased lipid peroxidation and macromolecular damage.
Aged rats that received GWE had significantly higher CAT
enzyme activity, perhaps due to ginsenosides, trace elements,
antioxidant vitamins, and other active compounds found in P
ginseng [611].
We observed a significant decrease in the activity of Se-GPx
in the liver, kidneys, heart, and lungs of aged rats. The
reduction in Se-GPx activity in aged rats can be attributed to
the decreased availability of its substrate, GSH, and its
inactivation by superoxide radicals that accumulate because
of decreased SOD activity [15,3034]. In addition, excessive
ROS production can also reduce the availability of Se, which is
an essential component of Se-GPx. Furthermore, Se concen-
tration can decrease gradually with advanced age [41].
Therefore, aged rats have lower Se-GPx activity compared
with young rats. Selenium is a micronutrient present in high
concentrations in certain food, including P ginseng, and forms
a vital part of GPx and various Se-dependent enzymes.
Specifically, GPx contains a selenocysteine residue in its
active site, which is essential for peroxidase activity. We
demonstrated that Se-GPx activity could be restored by GWE
treatment in aged rats; this might be due to the nutrients in P
ginseng such as ginsenosides, antioxidant vitamins, and trace
elements including selenium [611].
In this study, we found that GR activity was significantly
reduced in the liver, kidneys, heart, and lungs of aged rats
compared with young rats. Our results are consistent with
those presented in earlier reports [3234]. The decrease in GR
activity might lead to changes in the crucial ratio of GSH/
glutathione disulfide and cause oxidative damage to cells in
vivo. Nonetheless, we found increased GR activity in GWE-
treated aged rats, and this effect might be due to sulfur-
containing amino acids such as cysteine and methionine.
Both of these amino acids significantly reduce lipid peroxida-
tion and restore GSH, a substrate of GR. Thus, increased GSH
availability due to GWE treatment may also be responsible for
increased GR activity in aged rats.
 N U TR IT ION RE S E ARCH 3 2 ( 2 0 12 ) 71 8 7 2 6
Glutathione-S-transferases are a family of enzymes with
an important role in detoxification by catalyzing the conjuga-
tion of aging-associated free radicals with GSH. We observed
significantly reduced GST activity in the liver, kidneys, heart,
and lungs of aged rats. The diminished activity of GST in aged
rats can be attributed to the increased levels of ROS and
decreased availability of its substrate, GSH, and our results
confirm the data presented in earlier studies [15,30,34].
Oxidative stress is one of the most important mechanisms
behind age-related decrease in GST activity in aged rats. In
this study, GWE played an important role in improving GSH
level in rats, and this may be due to increased GST activity.
Reduced glutathione plays an important role in the
maintenance of the intracellular redox state and in the
cellular defense against oxidative damage. In aged rats, the
level of GSH was decreased in the liver, kidneys, heart, and
lungs, as was reported previously [15,3034]. The decrease in
GSH levels in the tissues of aged rats was completely
reversed with dietary supplementation with GWE. This
might be due to the ginsenosides (the active compounds of
P ginseng), which scavenge free radicals and restore GSH
levels. In addition, sulfur-containing amino acids such as
cysteine and methionine have thiol groups that can help to
synthesize GSH [42]. Furthermore, vitamin E and vitamin C
levels of aged rats were lower in the liver, kidneys, heart, and
lungs compared with young rats, which is consistent with
reports showing that the tissue levels of vitamin E and
vitamin C decrease with increased age [15,3034]. Higher
levels of vitamins E and C were observed in the tissues of
aged rats treated with GWE. This effect is due to antioxidants
such as vitamins, sulfur-containing amino acids, trace
elements, and other active compounds of P ginseng [611]
that scavenge free radicals and restore vitamin E, vitamin C,
and GSH levels.
In conclusion, our findings suggest that GWE can prevent
lipid peroxidation by enhancing enzyme and nonenzyme
antioxidant defense systems in aged rats. The skewed
oxidant-antioxidant balance in senescence can be attributed
to antioxidant nutrient insufficiency. However, the nutrients
in GWE, including ginsenosides, vitamins, sulfur-containing
amino acids, trace elements, and other active compounds,
appear to help maintain the antioxidant status of the cell.
Our data illustrate the potential of GWE in preventing age-
related oxidative damage in rats and suggest that GWE has
potential as a therapy for attenuating the pathological
consequences of aging.
Acknowledgment
This research was supported by the 2010 grant from the
Korean Society of Ginseng.
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