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Althea Medical Journal. 2014;1(2)
100 AMJ December, 2014
The Analgesic Effect of Pineapple Fruit Juice on Mice
Ainul Atiqah binti Hilmi¹, Vycke Yunivita KD², Henny Anggraini Sadeli³
1Faculty of Medicine, Universitas Padjadjaran, 2 Department of Pharmacology and Therapy, Faculty
of Medicine, Universitas Padjadjaran, 3 Department of Neurology, Faculty of Medicine, Universitas
Padjadjaran/Dr.Hasan Sadikin General Hospital
Abstract
Background: Pain is a feeling stimulated by the nervous system which can be suppressed by giving an
analgesic agent. Some studies revealed that pineapples have an analgesic effect. This study aim was to
determine analgesic effect of pineapple on mice.
Methods: In this experimental study, the effect was examined by using a writhing method on the 28 male
mice. Subjects were divided into 4 groups with 7 mice each. The control group received aquades and other
groups received pineapple fruit juice with 20%, 40% and 80% concentration with the dosage of 10 mL/
kg/body weight. After 30 minutes, 3% acetic acid was injected intraperitoneally to induce pain. Writhing
responseswere observed every 5 minutes for 30 minutes.
Results: The result for mean of total writhing reaction was 2.39±0.40, 1.92±0.40, 1.50±2.13, 1.66±0.11
mice with 20%, 40% and 80% concentration compared to control group (p=0.023;p=0.000 and p=0.000
respectively). Most optimal concentration was40% with the protective percentage equal to 71.8%.
Conclusion: Pineapple fruit juice concentrations (20%, 40%, and 80%has an analgesic effect with the most
optimal concentration of 40%. [AMJ.2014;1(2):100–4]
Key words: Analgesic, pain, pineapple fruit, writhing method
Efek Analgesik Jus Buah Nanas Pada Mencit
Abstrak
Latar Belakang: Nyeri adalah sensasi perangsangan sistem saraf yang dapat ditekan dengan pemberian zat
analgetik. Penelitian sebelumnya menemukan bahwa nanas memiliki efek analgesik. Tujuan penelitian ini
adalah untuk mengetahui efek analgesik nanas pada mencit.
Metode: Efek analgesik diperiksa dengan menggunakan metode “writhing“ pada 28 mencit jantan. Subyek
penelitian dibagi menjadi 4 kelompok masing-masing 7 ekor. Kelompok kontrol menerima aquades dan
kelompok lainnya menerima jus buah nanas secara oral dengan konsentrasi 20%, 40% dan 80% dengan
dosis 10 mL/kg/berat badan. Setelah 30 menit, asam asetat 3% disuntikkan intra peritoneal untuk
menginduksi nyeri. Respons “writhing” diamati setiap 5 menit selama 30 menit.
Hasil: Hasil rata-rata reaksi total ”writhing”adalah 2.39 ± 0.40, 1.92 ± 0.40, 1.50 ± 2.13, 1.66 ± 0.11 masing-
”writhing” pada tikus dengan 20%, 40 % dan konsentrasi 80% dibandingkan dengan kelompok kontrol
(p=0,023; p=0,000; dan p=0,000masing-masing). Konsentrasi optimal adalah 40% dengan persentase
pelindung sebesar 71,8%.
Simpulan: Konsentrasi jus buah nanas (20%, 40% dan 80%) memiliki efek analgesik dengan konsentrasi
paling optimal pada 40%. [AMJ.2014;1(2):100–4]
Kata Kunci: Analgesik, nyeri, nanas, writhing
Correspondence: Ainul Atiqahbinti Hilmi, Faculty of Medicine, Universitas Padjadjaran, Jalan Raya Bandung-Sumedang
Km.21, Jatinangor, Sumedang, Indonesia, Phone: +6281912725563, +60199578082, Email: ainul_1908@yahoo.com.my
Althea Medical Journal. 2014;1(2)
101
Introduction
Pain is a feeling stimulated by the nervous
system. It is a complex phenomenon that warns
an individual about some tissue damage which
currently is occurring or has occurred.1It
happens when there is a damage or injury, the
tissue will release some chemical mediators
which will stimulate pain sensation.2 This
sensation can be supressed by using analgesic
drugs such as paracetamol and ibuprofen.
Some plants also exhibit analgesic effects
such as pineapple (Ananascomosus). In
previous study , pineapple had been proved
as a plant that have an analgesic effect due
to the presence of bromelain.3 This active
compound which can be found in the root and
stem of pineapple, bromelain, is a proteolytic
enzyme which has many pharmacological
activities such as analgesic agent, anti-
tissue permeability of antibiotic drugs.4
In this study 3% acetic acid had been
used to induce pain sensation on mice.
Acetic acid is an irritation agent that when
injected intraperitoneally, it usually induces
hypersensitivity.5 The objective of this study
was to determine the 20%, 40% and 80% of
the pineapple fruit juice concentration which
has the protective result as an analgesic effect.
Methods
The study subject used in this experiment
were 28 healthy male mice (Musmusculus)
with body weight of 20–30 gram each which
then were divided into 4 equal groups.
The research materials in this study were
pineapple fruit juice as the intervention with
the concentration of 20%, 40% and 80% for
group 2, group 3 and group 4 respectively.
To make 20% concentration of pineapple
fruit juice, 20 gram pineapple was used. The
juice and added with 100 mL distilled water.
For concentration of 40% and 80%, 40 gram
and 80 gram of pineapple fruit were used
respectively and then the same procedure
was applied.
Other materials used were 3% acetic
acid as the pain stimulation and aquades as
placebo. All mice were left freely for one week
as an adaptation period in the laboratory with
optimal room temperature without any effort
of intervention. Prior to the experiment, all
mice must undergo a fasting period for 18
hours with free access of drinking water.6
Writhing method or abdominal contraction
method was used in this study to evaluate the
analgesic effect. For the experimental group,
each will receive 20%, 40% and 80% pineapple
fruit juice concentration respectively via
oral and for thecontrol group, aquades was
givenorally as a placebo. After 30 minutes,
30% acetic acid was injected intraperitoneally
to induce pain. Right after acetic acid was
given , writhing response was observed. The
reaction was observed for 5 minutes every
30 minutes right after giving acetic acid. The
abdominal contraction, stretching of the
body, retraction of the abdomen followed by
withdrawing of the lower limbs backward.7
Protective percentage in this study was
calculated by using the formula of 100% -
(total number of writhing in experimental
group / total number of writhing in control
group) x 100. For statistical analysis, analysis
of variance (ANOVA) was used to see whether
test then subjected to Tukeyy HSD multiple
comparison post test.
This experiment was done in November
2012 at the Animal Laboratory of Pharmacology
and Therapy, Faculty of Medicine, Universitas
Padjadjaran , Jatinangor, Bandung, Indonesia.
Results
In this study, the data was analysed by
and due to data skewness original data of
mean of total writhing reaction in mice was
transformed into a new data to meet the
criteria of parametric test ANOVA..The formula
recorded in table 1.
value of 0.000 (p<0.05). Post hoc test by using
can be found (Table 3).
All three experimental groups showed
writhing response comparing to control group
(Table 3). It means that the analgesic activity
group (p<0.05), while in three experimental
between group 2 and 3 (p<0.05).
Pineapple fruit juice with concentration
of 40% show ed the most protective effect as
an analgesic than other,which was 71.8 %.
Ainul Atiqah binti Hilmi, Vycke Yunivita KD, Henny Anggraini Sadeli: The Analgesic Effect of Pineapple
Fruit Juice on Mice
Althea Medical Journal. 2014;1(2)
102 AMJ December, 2014
Table 2 Difference Total Writhing Reaction Between Groups
Group 1 Group 2 Group 3 Group 4 P
Mean of total
writhing reaction
in mice
2.39 1.92 1.50 1.66 0.000
Then, it was followed by 80% concentration
which was 61.6% and the last one was 20%
pineapple fruit juice concentration which was
35.4% (Table 4).
Discussion
Based on its pathophysiology, pain can be
visceral and neuropathic pain.8 Somatic pain
is usually located at the peripheral, visceral
pain involves intra-abdominal organ, and
neuropathic pain involves afferent neural
pathway.8
This study was conducted to examine
the analgesic effect of the pineapple fruit
(Ananascomosus) juice on the mice models.
The analgesic reactions were measured by the
total number of writhing in the mice every 5
minutes for 30 minutes. The mean of total
writhing reaction in mice was decreasing
from the mean group until group 3 which
received 40% concentration of pineapple fruit
(Ananascomosus) juice and increased again
in the group 4 when the mice in this group
had 80% concentration of pineapple fruit juice
(Table 1). This condition showed that the
pineapple juice had a ceiling effect. Ceiling
effect is a condition where at certain dosage,
the drugs does not give any therapeutic effect
instead this condition can increase the side
effect.9Because of the ceiling effect, 80%
concentration of pineapple fruit juice show ed
decreasing analgesic effect.
control group and experimental groups which
received 20%, 40, and 80% pineapple fruit
juice concentration (Table 2 and 3). It revealed
that at all concentrations, pineapple fruit juice
showed its analgesic effect by reducing the
bradykinin synthesis. But there was only a
3 in the experimental groups. Almost all
inhibiting the activity of cyclooxygenase (COX)
synthesis.10 In establishing the analgesic effect,
bromelain will decrease the bradykinin level
released when tissue is damaged . Bradykinin
will stimulate the production of prostaglandin
while prostaglandin in turn will enhance the
sensitivity of bradykinin toward its receptor.2
When bradykinin is low, it also will decrease
11
Moreover, comparison between group 2
(20% concentration of pineapple juice) and
group 3 (40% concentration of pineapple
the protective percentage measurement, the
result show ed that pineapple fruit juice with
concentration of 40% ha d the most protective
result.
Previously, there had been studies upon
the effect of bromelain extract of pineapple
stems towards the analgesic reaction in mice.
The chemical constituent responsible for
analgesic effect is known as bromelain which
is a proteolytic enzyme. This active compound
Table 1 Mean of total writhing reaction in mice
Groups Mean of total writhing reaction in mice SD
1 2.39 0.40
2 1.92 0.26
3 1.50 2.13
4 1.66 0.11
Note:
Group 1 : given aquades
Group 2 : given 20% pineapple fruit juice concentration
Group 3 : given 40% pineapple fruit juice concentration
Group 4 : given 80% pineapple fruit juice concentration
Althea Medical Journal. 2014;1(2)
103
Table 3 Post hoc test by Tukeyy HSD method
Group
Comparison P SD
1 2 0.023
1 3 0.000
1 4 0.000
2 3 0.042
2 4 0.310
3 4 0.696
Note:
Group 1 : given aquades
Group 2 : given 20% pineapple fruit juice concentration
Group 3 : given 40% pineapple fruit juice concentration
Group 4 : given 80% pineapple fruit juice concentration
is present in the stem as well as in the fruit.
From the data in this study, we can conclude
that the pineapple fruit also has an analgesic
activity like the stem. But to ensure the safety
issue, several studies should be addressed to
reveal the side effect, toxicity, minimum and
maximum dosage of pineapple fruit as an
analgesic. Other studies can also be conducted
to compare the pineapple fruit to other non–
opioid analgesic that are commonly used in
the community such as paracetamol and
ibuprofen
References
1.
type of pain.[Online Web Page] 2011 [cited
2012 April 15]. Available from: http://
www.healingchronicpain.org/content/
2. Bueno L, Fioramonti J. Visceral perception:
3.
analgesic effect of bromelain in mice and
rats.Universa Medicina. 2005;155–60.
4. Bhattacharyya BK. Bromelain: an Overview.
5. Bueno L, Fioramonti J, Garcia-Villar R.
Visceral afferent pathways: a source of
new therapeutic targets for abdominal
6. Smith A. Fasting in rodents.
NorecopaVeterinaer Institute; 2009[cited
2012 May 8]. Available from: http://www.
norecopa.no/norecopa/vedlegg/Food-
deprivation.pdf.
7. Sureshkumar S, Sivakumar T, Chandrasekar
M, Suresh B. Investigating the anti-
leaves of wedeliachinensis (osbeck) merr.
In standard experimental animal models.
8. Cole BE. Pain management: classifying,
understanding, and treating pain. Hosp
9. Aschenbrenner DS, Venable SJ. Drug in
nursing. 3rd ed. Philadelphia: Lippincott
William & Wilkins; 2009.
10. Vane JR, Botting RM. The mechanism of
11. Brien S, Lewith G, Walker A, Hicks SM,
Middleton D. Bromelain as a treatment for
Table 4 Protective percentage against 3% acetic acid
Group Total writhing Protective percentage (%)
Aquades 206 0
20% pineapple juice 133 35.4
40% pineapple juice 58 71.8
80% pineapple juice 74 61.6
Ainul Atiqah binti Hilmi, Vycke Yunivita KD, Henny Anggraini Sadeli: The Analgesic Effect of Pineapple
Fruit Juice on Mice
Althea Medical Journal. 2014;1(2)
104 AMJ December, 2014
osteoarthritis: a review of clinical studies.
Evid Based Complement Alternat Med.
