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Analgesic and anti-inflammatory activities of

[6]-gingerol

Article in Journal of Ethnopharmacology February 2005

DOI: 10.1016/j.jep.2004.09.009 Source: PubMed

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 Journal of Ethnopharmacology 96 (2005) 207210

Analgesic and anti-inflammatory activities of [6]-gingerol

Haw-Yaw Younga , Yen-Lin Luob , Hao-Yuan Chengc , Wen-Chiuan Hsiehc ,
Jung-Chun Liaoc , Wen-Huang Pengc,
a TzuHui Institute of Technology, 926 PingTung, Taiwan, ROC
b Institute of Human Resource Management, National Sat Yat-San University, 804 Kaohsiung, Taiwan, ROC
c Institute of Chinese Pharmaceutical Sciences, China Medical University, 404 Taichung, Taiwan, ROC

Received 1 October 2003; received in revised form 6 July 2004; accepted 6 September 2004

Abstract

In the present study, the analgesic and anti-inflammatory effects of [6]-gingerol, which is the pungent constituent of ginger, were performed.
Intraperitoneal administration of [6]-gingerol (25 mg/kg50 mg/kg) produced an inhibition of acetic acid-induced writhing response and
formalin-induced licking time in the late phase. [6]-Gingerol (50 mg/kg100 mg/kg) also produced an inhibition of paw edema induced by
carrageenin. These results suggested that [6]-gingerol possessed analgesic and anti-inflammatory activities.
2004 Published by Elsevier Ireland Ltd.

Keywords: [6]-Gingerol; Formalin; Writhing response; Anti-inflammation

1. Introduction 2. Material and methods

Herbal medicines are increasingly used by the general 2.1. Chemicals

population (Austin, 1998), Zingiber ofcinale ROSCOE
(family Zingiberaceae) is one of the most frequently used
ingredients in Chinese folk medicine for treatment of vari-
ous infectious disease. Gingerols, the active components of
ginger, represent a potential new class inhibitor of platelet
activators. (Koo et al., 2001). [6]-Gingerol has been found
to possess a variety of interesting pharmacological effects,
for example, antipyretic, cardiotonic effects, and inhibition
of spontaneous motor activity and prostaglandin biosynthesis
(Suekawa et al., 1984; Kiuchi et al., 1992). [6]-Gingerol has
been used as a marker substance of ginger. The [6]-gingerol
contents in different preparations were evaluated (Young and
Chen, 2002). However, little information is availabile on
the analgesic and anti-inflammatory effects of [6]-gingerol.
Therefore, we examined the effects of [6]-gingerol on Noci-
ception induced by acetic acid and formalin in mice and paw
edema induced by -carrageenin (CARR) in rats.
Corresponding author. Fax: +886 4 24075683.
E-mail address: whpeng@mail.cmu.edu.tw (W.-H. Peng).
[6]-Gingerol used in this study was purchased from
WAKO company (Japan). The company guaranteed batch-
to-batch chemical consistency by high performance liquid
chromatograph and biological reliability by lymphocyte
proliferation assay. Its purity was more than 99%. Acetic
acid and formalin were purchased from Darmstadt (F.R. Ger-
many). Carrageenin, indomethacin, aspirin were obtained
from Sigma (St. Louis, MO, USA).
2.2. Animals
Male ICR mice (18 g22 g) and female Wistar rats
(180 g240 g) were obtained from the National Labora-
tory Animal Breeding and Research Center, National Sci-
ence Council, Taiwan. They were housed in standard cages
at a constant temperature of 22 1 C, relative humidity
55 5% with 12 h darklight cycle for at least 1 week be-
fore the experiment. They were fed with food and water ad
libitum.

0378-8741/$ see front matter 2004 Published by Elsevier Ireland Ltd.

doi:10.1016/j.jep.2004.09.009
208 H.-Y. Young et al. / Journal of Ethnopharmacology 96 (2005) 207210
All experimental procedures were performed according
to the NIH Guide for the Care and Use of Laboratory Ani-
mals. The choice of dosage was based on the recommendation
printed on the pamphlet to be taken by humans. The placebo
groups were intraperitoneal with 0.2 ml saline. A bent blunted
27-gauge needle connected to a 1 ml syringe was used for the
intraperitoneal administration. All tests were conducted un-
der the guidelines of the International Association for the
Study of Pain (Zimmermann, 1983).
2.3. Acetic acid-induced writhing response
Male ICR mice were used in groups of eight ani-
mals per dose of drugs. The animals were pretreated with
[6]-gingerol (12.5 mg/kg, 25 mg/kg, 50 mg/kg, i.p.) or in-
domethacin (10 mg/kg, i.p.) for 30 min or 20 min, respec-
tively, prior to intraperitoneal injection of 1% acetic acid
(0.1 ml/10 g). Five minutes after the i.p. injection of acetic
acid, the number of writhings during the following 10 min
was counted. Control mice received normal saline (Koster et
al., 1959; Taber et al., 1969).
2.4. Formalin test
Male Albino ICR mice (eight per group), 18/25 g, were
kept in plexiglas cages with free access to food and water.
Testing took place in the middle of the light period of a 12-h
light:12-h dark cycle. Each animal was tested once only. The
[6]-gingerol (25 and 50 mg/kg) and indomethacin (20 mg/kg)
were suspended in tween 80 plus 0.9% (w/v) saline solu-
tion and administered i.p. in a volume of 0.18/0.2 ml. Con-
trol group received only drugless vehicle (0.18/0.2 ml). The
antinociceptive activity of the drugs was determined using
the formalin test described by Dubuisson and Dennis (1977).
One hour before testing, the animal was placed in a standard
cage (30 cm 12 cm 13 cm) that served as an observation
chamber. Twenty microlitres of 1.0% formalin injected to the
dorsal surface of the right hind-paw. The mice were observed
for 40 min after the injection of formalin, and the amount of
time spent licking the injected hind-paw was recorded. The
first 5 min post formalin injection is known as the early phase
and the period between 15 min and 40 min as the late phase.
The drugs were administered 30 min before injection of for-
malin. The total time spent on licking or biting the injured
paw (pain behavior) was measured with a stoped watch. The
activity was recorded in 5 min interval.
2.5. Carrageenan-induced rat paw edema
The anti-inflammatory activity of [6]-gingerol was deter-
mined by the carrageenan-induced edema test in the hind
paws of rats. Male Albino Wistar rats (eight per group),
180 g250 g, were fasted for 24 h before the experiment with
free access to water. Fifty microlitres of a 1% suspension
of carrageenan (Sigma Co., USA) in saline was prepared
30 mins before each experiment and was injected into the
plantar side of right hindpaws of the rats. The [6]-gingerol
and indomethacin were suspended in tween 80 plus 0.9%
(w/v) saline solution. The final concentration of tween 80 did
not exceeded 5% and did not cause any detectable effect per
se. The [6]-gingerol at the doses of 50 mg/kg, 100 mg/kg and
250 mg/kg and indomethacin at the doses of 10 mg/kg were
administered intraperitoneally (i.p.). Drugs or drugless ve-
hicle were injected 30 min before the carrageenan treatment.
Paw volume was measured immediately after carrageenan in-
jection and at 0.5-, 1-, 1.5-, 2-, 2.5-, 3-, 3.5-, 4-, 4.5- and 5-h
intervals after the administration of the edematogenic agent
using a plethysmometer (model 7159, Ugo Basile, Varese,
Italy). The degree of swelling induced was evaluated by
the ratio a/b, where a and b are the volume of the right
hind paw after and before carrageenan treatment, respec-
tively. Indomethacin was used as a positive control compound
(Mascolo et al., 1989). The right paw edema was measured
with a plethysmometer (Ugo Basile) 5 h after -carrageenin
application.
2.6. Statistical analysis
Data are expressed as mean S.E.M. Statistical eval-
uation was carried out by one-way analysis of variance
(ANOVA). Statistical significance is expressed as * P < 0.05
or ** P < 0.01, *** P < 0.001.
3. Results and discussion
Fig. 1 shows the pain behavior of writhing response which
is presented as cumulative abdominal stretching response.
The treatment of animals with [6]-gingerol (25 mg/kg,
50 mg/kg) produced a significant and dose dependent inhi-
bition of the control writhes. The inhibition by [6]-gingerol
Fig. 1. The effects of [6]-gingerol administered intraperitoneally on
1% acetic acid-induced writhing response in mice. Data represented as
mean S.E.M. (n = 8). *** P < 0.001 as compared with the control group.
 H.-Y. Young et al. / Journal of Ethnopharmacology 96 (2005) 207210
Fig. 2. The effects of [6]-gingerol administered intraperitoneally on the early
phase and late phase in 20 l of 1% formalin test in mice. Data represented as
mean S.E.M. (n = 8). *** P < 0.001 as compared with the Formalin group.
(25 mg/kg) was similar to that produced by indomethacin
(10 mg/kg). The acetic writhing test is normally used to study
the peripheral analgesic effects of drugs. Although this test is
a nonspecific (e.g., anti-cholinergic, antihistaminic and other
agents also show activity in the test), it is widely used for
analgesic screening and involves local cholinergic and his-
taminic receptors, and the mediators acetylcholine and his-
tamine (Shibata et al., 1989). This result indicates that the
analgesic effect of [6]-gingerol might be mediated by its pe-
ripheral effect.
[6]-Gingerol (25 mg/kg, 50 mg/kg) produced significant
(P < 0.001) inhibition in the late phase of formalin induced
pain, respectively (Fig. 2). The positive control indomethacin
(20 mg/kg) also produced significant (P < 0.001) inhibition
in the late phase. The formalin test is a valid and reliable
model of nociception and is sensitive for various classes of
analgesic drugs. Formalin test produced a distinct biphasic
response and different analgesics may act differently in the
early and late phases of this test. Therefore, the test can be
used to clarify the possible mechanism of antinociceptive ef-
fect of a proposed analgesic (Tjolsen et al., 1992). Centrally
acting drugs such as opioids inhibit both phases equally
(Shibata et al., 1989) but Peripherally acting drugs such
as aspirin, indomethacin and dexamethasone only inhibit
the late phase. The late phase seems to be an inflammatory
response with inflammatory pain that can be inhibited by
anti-inflammatory drugs (Hunskaar and Hole, 1987; Rosland
et al., 1990). The effect of [6]-gingerol on the late phase of
formalin test suggests that its activity may be resulted from
its peripheral action when compared with indomethacin
activity in this respect. Based on the results of this study, we
suggest that the antinociceptive effect of [6]-gingerol may
be attributed to inhibition of prostaglandin release and other
mediators involved in this test (Farsam et al., 2000).
209
Fig. 3. The effects of [6]-gingerol (Gin) and indomethacin (Indo) on rats
hind paw edema induced by -carrageenin (CARR). Data represented as
mean S.E.M. (n = 8). ** P < 0.01, *** P < 0.001 as compared with the CARR
group.
As shown in Fig. 3, -carrageenin induced the paw edema.
Both of [6]-gingerol (50 mg/kg, 100 mg/kg, 250 mg/kg)
and indometacin (10 mg/kg) significantly decreased the
-carrageenin induced the paw edema (P < 0.001). The
carrageenan test is highly sensitive to nonsteroidal anti-
inflammation drugs, it has long accepted as a useful
phlogistic tool for investigating new anti-inflammatory
drugs (Just et al., 1998). The degree of swelling of the
carrageenan-injected paws was maximal 3 h after injection
and the mean increase in volume at that time was about 100%
(a/b: 2.0) in control group. Statistical analysis shows that the
edema inhibition of [6]-gingerol at the doses of 50 mg/kg,
100 mg/kg and 250 mg/kg are significantly different from
control group (P < 0.001).
The results showed that [6]-gingerol at the dose of 50 and
100 mg/kg had more edema inhibition than indomethacin at
the doses of 10 mg/kg (P < 0.01). Results indicated that sim-
ilar activity against carrageenan-induced rat paw edema was
observed with [6]-gingerol at the dose of 100 mg/kg and
indomethacin at the dose of 10 mg/kg. In this study, [6]-
gingerol exhibited a considerable anti-inflammatory activity
(ED50 = 85.32 mg/kg) in comparison with drug reference.
In inflammatory reactions, activated neutrophils re-
lease mediators such as platelet-activating factor, lysozyme
(Weissmann et al., 1980). These substances lead to bronchial
muscle contraction, vasodilation, increased vascular perme-
ability and inflammatory disease (Adams and Lichtenstein,
1979; Schwartz and Austen, 1984). Neutrophils also release
active substances such as hydrogen peroxide, superoxide an-
ions and hydroxyl radicals which play important roles in
producing cellular damage and are associated with aging
and rheumatism (Alien et al., 1989; Semb et al., 1990). [6]-
Gingerol has been found to possess substantial antioxidative
activity as determined by inhibition of phospholipid perox-
idation induced by the FeCl3 -ascorbate system. The actual
 210 H.-Y. Young et al. / Journal of Ethnopharmacology 96 (2005) 207210
anti-inflammatory mechanism of [6]-gingerol was worthy of
further study in the future.
In conclusion, the data obtained in this study demonstrated
that [6]-gingerol might have analgesic and ant-inflammatory
activities. Further studies are necessary to elucidate the mech-
anisms behind its traditional effects.
Acknowledgements
We are thankful to the China Medical University for
the financial support of this manuscript under contract No.
CMC91-CPS-03, CMC92-CPS-03.
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