Advances in Health Sciences Research, volume 20

1st International Conference on Community Health (ICCH 2019)

Anti-Inflammatory Effects of Kaempferia galanga

L. Rhizome Extract in Carrageenan-Induced
Female Rats

1st Galih Samodra 2nd Dina Febrina

Pharmacy Study Program, Faculty of Health Science Pharmacy Study Program, Faculty of Health Science
Harapan Bangsa University Harapan Bangsa University
Purwokerto, Indonesia Purwokerto, Indonesia
galihsamodra@uhb.ac.id dinafebrina@uhb.ac.id

Abstract— Kencur (Kaempferiae galanga L.) is a plant that is get rid of dirty blood and get through periods smoothly.
traditionally used for the treatment of various diseases Previous studies reported that the extract of kencur has
including inflammation. This study aimed to know the analgesic and anti-inflammatory effects [3], nematicidal
comparison of the effectiveness of kencur extracts with agents [4], mosquito repellent and larvacidal effect [5],
diclofenac sodium as an antiinflammatory agent. Fifteen vasorelaxants [6], antineoplastics [7], antioxidants [8], and
female rats aged 2-3 months (200250 gram) were divided into 5
antimicrobials [9].
groups: rats in negative control group only given a 2%
suspension of PGA, rats in positive control group given 20 One of the plants that has an anti-inflammatory effect is
mg/Kg diclofenac sodium, and rats in three different dose kencur (Kaempferia galanga L.). The choice of appropriate
groups given galanga rhizomes extract of 45, 90, and 180 extraction solvent is also important to obtain the best result
mg/Kg, respectively. All drugs were given 1 hour orally before of extraction efficiency and to obtain compounds that have
carrageenan injection of 1% as induction of edema. Edema pharmacological activities. The best solvent is ethanol or
volume was measured every 30 minutes to 390 minutes to its mixture with water because it is a good extraction
calculate the percentage of edema inhibition. The data were
analyzed using one-way ANOVA and followed by LSD test to
solvent for almost all low-molecular-weight compounds
see differences among groups. Ethanol extract of galanga such as saponins and flavonoids. The type of extraction
rhizome in the 5th hour at doses of 45, 90 and 180 mg/kg solvent also affects the number of active compounds in the
showed significant differences in a positive control group. extract based on the principle of “like dissolve like”, in
Each Sig. value was 0.002; 0.004; 0.002. This showed that the which polar compounds will dissolve in polar solvents and
ethanol extract of kencur had anti-inflammatory activity at all non-polar compounds will dissolve in non-polar solvents
dose levels. [10]. Ethanol is often used as a solvent in the laboratory
because it is an inert solvent in which it does not react with
Keywords: Kaempferia galanga L., anti-inflammatory, other components [11].
carrageenan In this study, the animals tested were rats because
metabolism in the rats’ body and humans are physiologically
I. INTRODUCTION more similar and the structure of the tissue in the rats’ feet is
Inflammation is linked to certain diseases that can be easier to be treated with subcutaneous induction compounds.
found in the community, usually characterized by swelling, Methanol and ethanol are both alcohol derivative solvents
pain, redness, and heat. The inflammation can be caused by which have hydroxyl (OH) groups and have carbon chains
a normal protective response to tissue injury from physical (atomic C). The number of C atoms in ethanol is greater
trauma, hazardous chemicals, or microbiological agents. (two C atoms), and thus this compound can dissolve
Inflammation is a response of the body's efforts to activate secondary metabolites that are non-polar in purslane plants.
or destroy invading organisms, remove irritants and prepare Other studies also mentioned that kencur rhizome extract at
phases of the tissue repair process [1]. a dose of 45 mg/kg was able to show a very high percentage
of inhibition compared to other dose groups and positive
Kencur (Kaempferia galanga L.) is one of the herbal control (51.27%) [12]. In this case, society has lack of
plants of the Zingiberaceae family which has many information about Kaempferia galanga extract that functions
pharmacological activities [2]. Society in Indonesia as an antiinflammatory drug. This extract has a great
traditionally uses kencur as medicine for inflammation of potential, then society should exploit it. Based on this study,
the stomach (gastritis), inflammation of the child's ear (ear
it is necessary to test various methods and comparisons.
infection), influenza in infants, colds, headache, cough,
diarrhea, sore eyes, sprains, and fatigue. It is also used to

Copyright © 2020 The Authors. Published by Atlantis Press SARL.

This is an open access article distributed under the CC BY-NC 4.0 license -http://creativecommons.org/licenses/by-nc/4.0/. 13
 Advances in Health Sciences Research, volume 20

II. METHODS 2. Rats were acclimatized for 1 week to adapt to the

experimental environment.
Anti-inflammatory test
3. Before the experiment, rats were deprived of food
A. Preparation for Animal experiment for ± 18 hours but still given drink.
4. At the beginning of the research, all rats were
Before being used, rats were acclimatized for 1 marked with markers at the ankle and therefore the
week in a cage to adapt to a new environment. All rats foot was always the same when entered into the
were kept in the same conditions, given the same food mercury.
and drink, and observed their general condition 5. The initial volume of rats’ feet (paw) was measured
regularly, and weighed. Before the experiment, rats before being treated and it represented the base paw
were deprived of food for ± 18 hours but they still volume (V0).
received water ad libitum [13]. Rats that were sick 6. The negative control group was given a 2%
which had standing fur, were less active and had suspension of PGA, while the positive control
unclear eyes were excluded in this study. Fifteen rats group was given a suspension of diclofenac sodium
were used and randomly divided into 5 groups at a dose of 20 mg/kg. The other three groups were
consisting of 3 rats of each (referring to WHO given a suspension of the compound test according
provisions). The following is the description of the to the planned doses given orally.
treatment given to each group of rats: 7. One hour later rats were injected 0.1 ml of 1%
carrageenan by subplantar injection in the rats left
TABLE 1. GROUP ANIMAL EXPERIMENT
Number leg. Before the carrageenan was injected, the area
Group
of rats
Treatment of the rats’ paw was wiped with alcohol swabs.
8. Furthermore, edema volume was measured in the
Negative 3 Have given the 2% PGA
30th, 60th, 90th, 120th, 150th, 180th, 210th, 240th,
control suspension + induced with 1% 270th, 300th, 330th, 360th, and 390th minute after
group carrageenan as much as 0.1 induction using plethysmometer represented as the
mL final volume (Vt).
Positive 3 Have given sodium diclofenac 9. The percentage of edema and average (mean)
control at a dose of 20 mg/Kg in 2% percentage of edema inhibition was calculated
using the following formula:
group PGA + induced with 1%
carrageenan as much as 0.1 Description :
mL Vt: paw volume of rats at time t (after carrageenan-
Dosage I 3 Have given a suspension of induced) V0: paw volume of rats at time 0 (before
test compound extract in 2% carrageenan-induced)
PGA (dose 45 mg/Kg) +
induced with 1% carrageenan
as much as 0.1 mL Description :
a : % edema in negative control group b : % edema in
Dosage 3 Have given a suspension of
test group [14]
II test compound extract in 2%
PGA (dose 90 mg/kg) +
induced with 1% carrageenan III. RESULTS AND DISCUSSION
as much as 0.1 mL In this study, the anti-inflammatory activity test of
Dosage 3 Have given a suspension of kencur rhizome extract was done by the Winter method
(carrageenan induction). Carrageenan induction is the
III test compound extract in 2%
possible method to be analyzed after given a single dose of
PGA (dose 180 mg/kg) + nontoxic in one day. Besides, this method is the most widely
induced with 1% carrageenan used as it is simple, easy, and suitable for testing the anti-
as much as 0.1 mL inflammatory activity of a compound in small amounts [15].
The use of carrageenan as edema induction in rats’ feet
B. Anti-inflammatory Activity Test using Carrageenan- has been widely used in testing the anti-inflammatory
Induced Rats Paw Method (Winyard & Willoughby, activity of a drug compounding [16; 17]. Edema formed by
carrageenan induction is in the form of acute inflammation
2003; Winter et al, 1962; Morris, 2003)
[18; 19]. Carrageenan was chosen in this study because it
The followings are the procedure of the experiment. can trigger the release of prostaglandins after being injected
1. Rats were weighed and grouped randomly into 5 in rats and therefore this compound can be used to find
groups. Each group consisted of 3 rats.

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antiinflammatory drugs to inhibit prostaglandin synthesis significant edema and there were differences among test
[15]. Rats weighed 150-250 grams. groups. The increase in edema volume was caused by the
In this experiment, the animals tested were grouped release of inflammatory mediators such as prostaglandins,
into 5 in which each consisted of 3 rats (referring to WHO histamine, bradykinin, and serotonin in tissues after
provisions). The experimental group consisted of a negative carrageenan induction. Besides, the decrease in edema
control group (2% suspension of PGA), a positive control volume of each test group at the fifth hour was due to the
group (diclofenac sodium suspension at a dose of 20 mg/kg), effect of carrageenan which had begun to decrease. The
and dose test groups with varying doses of 45 mg/kg, 90 formation of edema by carrageenan only lasts for 5-6 hours
mg/kg and 180 mg/kg. and gradually decreases within 24 hours after injection [20].
The choice of dose varied referring to the research of Furthermore, the percentage of edema formation from
Hasanah [12], in which the extract of kencur rhizome has the the edema volume of the rats’ paw can be calculated to see
most optimal anti-inflammatory activity at a dose of 45 the differences in edema between groups by using this
mg/kg. Tests were carried out using 3 dose variations, formula: (Vt - Vo) / Vo x 100%), which is Vt = Volume of
indicating the greater the dose given, the greater the edema every hour and V0 = volume rats’ paw before the
antiinflammatory activity produced. The results showed a treatment. The results of the calculation of the mean
significant percentage of inhibition in treatments at a dose of percentage of edema on the rats’ paw can be described as
45 mg/kg and the percentage of inhibition was 51.27%. follows:
Before the treatment, the basic volume of rats’ foot (V0) was
measured first and furthermore, rats’ foot volume was
measured every 30 minutes for 6.5 hours and it represented
Vt.
The anti-inflammatory activity of a compound can be
seen from its ability to inhibit the formation of edema in the
rats’ paw [19]. This was assessed from the increase or
decrease in edema volume every 30 minutes. The edema
volume was measured using a plethysmometer. The use of
this tool required accuracy in the measurement because the
volume of mercury always have to be the same in every test.
Rats’ feet were marked on the ankle joint and therefore the
immersion of rats’ feet into the mercury remained the same Figure 2. The graph of the relationship between mean percentages of
every 30 minutes. The edema volume of rats’ feet was edema and time (minute)
measured every 30 minutes for 6.5 hours. After carrageenan
induction, the mean volume of edema was obtained and it From the graph above, it can be seen that the negative
can be illustrated in the following figure: control group has a very high percentage of edema
formation and has a significant increase continuously to the
3rd hour. Then it begins to fall at the 4th hour and rises
again to the 6.5th hour. In this case, animals tested were
only given a suspension of PGA that was unable to inhibit
the formation of edema and the response to edema only
relies on rats’ immunity. Compared to the positive control
group, the dose I, dose II and dose III of the edema volume
were lower. Based on the results of the statistical tests, the
four groups were significantly different compared to the
negative control group (ρ> 0.05). Thus, it can be concluded
Figure 1. The edema volume in the 30th to 390th minutes. that these four groups have antiinflammatory activity in
inhibiting the formation of edema in the rats’ paw.
From the graph of mean edema volume above, it can be Furthermore, the mean percentage of edema formation
seen the difference between the negative control group and inhibition in each group can also be calculated to see the
the treatment group. The negative control group that was not anti-inflammatory activity. It can be illustrated in figure 3.
given extracts and drugs has a significant increase compared
with other treatment groups. The group which was not
induced by carrageenan did not form edema at all and rats’
feet do not show an increase or a decrease in edema volume.
Unlike the group with no induction, the groups with
carrageenan induction show an increase or a decrease in
edema volume after induced by carrageenan. This shows
that 0.1 mL of 1% carrageenan-induced could form

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in each group. The result showed a significant difference

between the positive control group and the 45, 90, and 180
mg/kg dose groups. This shows that the ethanol extract of
kencur with doses of 45, 90, and 180 mg/kg has an anti-
inflammatory.
The edema formation process induced by carrageenan
occurs in two phases and involves a variety of inflammatory
mediators [22]. The first phase occurs for 3 hours after
carrageenan induction in which histamine, serotonin,
bradykinin are released and have an increase in
prostaglandin synthesis around the injured tissue. The
second phase occurs from the fourth hour to the fifth hour
and there is an absorption of prostaglandins, proteases and
Figure 3. The graph of the relationship of the mean percentages of edema
inhibition and time (minute) lysosomes [22; 23; 24]. Generally, this second phase is
sensitive to anti-inflammatory drugs [25].
From the graph above, it can be seen that the In this study, the drug used in positive control was
percentage of edema inhibition of each group is very diclofenac sodium at a dose of 20 mg/kg showing the best
different. It appears that the positive control group with percentage of edema inhibition. This is consistent with the
diclofenac sodium drug increases continuously, although the literature that diclofenac sodium as an NSAID inhibits the
effects in the first 60 minutes are still low. nonselective cyclooxygenase enzymes, thereby it inhibits
Based on the graph, the dose I group is lower compared the release of prostaglandins [26]. Of the three dose groups,
to the positive control in the 30th minute to the 390th the dose II group (90 mg/Kg) has a high average percentage
minute, although in the 120th and 360th minute the dose I of inhibition compared to the other dose groups. In addition,
group is higher. The dose II group has a fluctuating the dose I and III (45 mg/kg and 180 mg/kg) have the same
percentage of edema inhibition from the 30th minute to the average percentage of inhibition. From this study, a decrease
390th minute. The dose II group decreases in the 120th to in the dose of kencur rhizome extract compound increased
the 330th minute, then increases to the 390th minute. edema inhibition ability in the rats’ paw. This is because
Furthermore, the dose III group has a fluctuating percentage actually several types of drugs in high doses cause the
of inhibition from the first to the last minute. The highest release of histamine directly from mast cells causing blood
average percentage of inflammation inhibition is diclofenac vessels to be more permeable to plasma fluid and causing
sodium starting in 30 minutes after treatment and reaches its inflammation processes (immunological processes occur)
peak at 390 minutes. The smallest percentage of [27].
inflammation inhibition is at a dose of 45 mg/kg. Kencur
ethanol extract at a dose of 90 mg/kg has a potential effect
on edema inhibition. Certain ingredients can have anti- IV. CONCLUSIONS
inflammatory effects. It can be proved that swelling reduced Ethanol extract of kencur rhizome extract in the 5th
by 50% or more when experimental animals were induced hour at doses of 45, 90 and 180 mg/kg showed significant
by carrageenan 1% [21]. differences in the positive control group and each Sig. value
The edema inhibition in each group was tested for its was 0.002; 0.004; 0.002. This showed that the ethanol extract
normality using the Kolmogorov Smirnov test. The data of kencur rhizome had anti-inflammatory activity at all dose
normality test was normally distributed. The homogeneity levels.
test was also applied which shows the value (p> 0.05). This
means the data varied homogeneously. The ANOVA test
was performed to find out significant differences between ACKNOWLEDGMENTS
the five groups. The ANOVA test result showed that Sig. The authors would like to express gratitude for the Dwi
value at the 5th hour was 0.006. It can be concluded that Puspita Educational Foundation of Harapan Bangsa
there were significant differences between the dose University which funded this research.
treatment groups and the positive control group. a further
test was conducted using LSD to determine the differences

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