J Acupunct Meridian Stud 2009;2(1):75−78

RE S E A RCH A RTI CL E

The Effect of Carum Copticum Extract on

Acetylcholine Induced Contraction in
Isolated Rat’s Ileum
Seyed Hassan Hejazian-Y1, Mohammad Hossein Dashti-R2*,
Seyed Majid Mahdavi3, Masood Anvar Qureshi4
1
Herbal Medicine Research Center, School of Medicine, Shahid Sadughi University of Medical Sciences
and Health Services, Yazd, Iran
2
Department of Physiology, School of Medicine, Shahid Sadughi University of Medical Sciences and
Health Services, Yazd, Iran
3
Department of Pharmacology, School of Medicine, Shahid Sadughi University of Medical Sciences and
Health Services, Yazd, Iran
4
Department of Physiology, Dow Medical College, Dow University of Health Sciences, Karachi, Pakistan

Received: Aug 05, 2008 Abstract

Accepted: Jan 13, 2009 Aims: There are many biological investigations for determining an effective cure
for the dysfunction of gastrointestinal tracts, using herbal medicine. It has been
KEY WORDS: reported that Carum Copticum is a bactericidal agent and possesses anticholinergic,
acetylcholine; antihistaminic and β-adrenergic stimulatory effects in some tissues. However, these
Carum Copticum; effects of Carum Copticum on mechanical activities of isolated intestine are not
clearly identified yet. The present study has been designed to find out the specific
contraction;
effects of Carum Copticum on mechanical activity of isolated rat’s ileum.
ileum
Materials and Methods: In this study rat’s ileum contraction was recorded through
an isolated tissue chamber in an organ bath by using isotonic transducer and oscil-
lographic device. The effect of Carum Copticum extract on acetylcholine induced
contraction in isolated rat’s ileum was evaluated.
Results: Our findings showed that 1% aqueous extract of Carum Copticum reduces
the basal contractile activity of rat’s ileum. The extract also reduced acetylcholine
induced contraction to 40% of its maximum response. The inhibitory action of Carum
Copticum extract on acetylcholine induced contraction was similar but slower than
that of atropine sulfate.
Conclusion: The results of this study showed an inhibitory effect of Carum Copticum
extract on acetylcholine induced contraction in rat’s ileum.

1. Introduction Europe. The major component of its fruit is essen-

tial oil which is composed of γ-terpinene, p-cymene,
Carum Copticum (L.) is a plant in Umbelliferae α-pinene, β-pinene, and other substances such as
family with a white flower and small brownish seeds. thymol and carvacol [1]. In the Iranian traditional
This plant commonly grows in Iran, India, Egypt and medicine several therapeutic effects including

*
Corresponding author. Department of Physiology, Shahid Sadughi School of Medicine, Booali St. Safaeyah, Yazd 8944157963, Iran.
E-mail: dashti_mh@yahoo.com

©2009 Korean Pharmacopuncture Institute

76
expectorant, diuretic, anti-vomiting, analgesic, anti-
asthma, anti-dyspnea, and anti-spasm, is postulated
for Carum Copticum fruits [2]. Despite the availa-
bility of modern medications the propensity towards
the traditional medications is growing throughout
the world [3] which needs scientific investigations
for evaluating the therapeutic effects of medicinal
plants and their mechanisms of actions. Some of
these biological investigations have been focused
on gastrointestinal dysfunctions. One of the most
important abnormalities in gastrointestinal tract is
diarrhea, which may be produced by some infec-
tions or increase in motility [4,5]. Carum Copticum
is traditionally used in treating many gastrointesti-
nal disorders such as indigestion, colic, dyspepsia
and diarrhea [6] and is suggested to be anti nocic-
eptive [7] as well. It has been found that Carum
Copticum is very effective against some bacteria
and its effect on digestive tract may be due to its
antibacterial activity [8]. In other hand there are
evidences indicating the relaxant effects of Carum
Copticum on smooth muscles [9,10] and in our pre-
vious study we found that Carum Copticum extract
reduces the motility of rat ileum [11]. Intestinal
motility is controlled by circulatory hormones, in-
trinsic and extrinsic nerves that release a variety
of transmitters. The most important excitatory
transmitter in gastrointestinal tract is acetylcho-
line which acts via muscarinic receptors in enteric
smooth muscle cell membranes [12]. This study
was conducted to understand the probable anti-
cholinergic effect of Carum Copticum fruit extract
on isolated rat’s ileum.
2. Materials and Methods
In this study male Albino rats living in the standard
environmental and feeding conditions were used
for isolation of ileum. For experimental procedure
permission of the animal ethics committee of Shahid
Sadughi Medical University (Yazd, Iran), in accord-
ance with the internationally accepted principles
for laboratory animal use and care mentioned by the
European Community guidelines were obtained.
For extraction preparation, 40 g air-dried fruits
of Carum Copticum (identified by botanists in the
herbarium of Yazd herbal drug research center and
a sample specimen was stored and coded in this
center) were gently grounded and mixed with 250 mL
of double distilled water (DDW) and subjected to
steam distillation for 3 hours at atmospheric pressure.
Experiments were performed as described in
previous reports [13,14]. Adult male rats weighing
200−250 g were sacrificed by cervical dislocation.
Segments of ileum (2 cm in length), were obtained,
flushed of their contents and trimmed of mesentery.
S.H. Hejazian-Y et al
Preparations were suspended from the transducer
lever in the axis of the longitudinal muscle with
fine thread, mounted in the 50 mL internal chamber
of organ bath containing Tyrode’s solution, main-
tained at 37ºC and bubbled with 95% O2 and 5% CO2
[13]. Then, it was allowed to stabilize for 60 min-
utes prior to drug addition. Isotonic responses were
recorded using an isotonic transducer (T2) and an
oscillograph recording system (the bioscience 400
Series Washington oscillograph). Acetylcholine (Sigma
Aldrich Chemie Gmbh, Germany) was used as a stan-
dard contractile control at the end of each assay.
After recording a base line as a control for ideal
mechanical activity, the specimens were influenced
by different concentrations of Carum Copticum ex-
tract in a cumulative manner to yield the minimum
effective concentration which was 1% of total ex-
tract diluted in Tyrode’s solution. The effects of 1%
Carum Copticum extract alone, after and before
the infusion of acetylcholine (5 × 10−4 M) was assessed
on the specimens.
The specimens were also influenced by 5 × 10−6 M
atropine sulfate (Sigma Aldrich Chemie Gmbh) fol-
lowing the acetylcholine infusion as a standard
cholinergic antagonist. For each drug assay five sets
of experiments were done.
The data were calculated as mean changes in
specimen length from base line due to changes in
contractile activity and analyzed by paired Student’s
t-test. The statistical significance was considered
as p ≤ 0.05.
3. Results
Our findings showed that the Carum Copticum ex-
tract itself declines the amplitude of base line con-
tractions, in all five sets of experiments (3 ± 0.57 mm
vs. 4.28 ± 1.11 mm) which this relaxant effect was
significant (p = 0.022). The effective dose of acetyl-
choline conversely significantly (p = 0.000) elevated
the amplitude of base line contractions in all as-
sessments (19.57 ± 4.6 mm vs. 4.28 ± 1.11 mm). After
pretreating the specimens with 1% Carum Copticum
extract, the effective concentrations of acetylcho-
line only led to about 40% of its maximum response
(Figure 1).
The contractile effects of acetylcholine were
diminished by the infusion of atropine sulfate or
Carum Copticum extract.
The inhibitory effect of atropine sulfate (10−4 M)
on acetylcholine induced contraction was observed
after a few second and completed in 30 seconds,
while the inhibition due to 1% Carum Copticum ex-
tract was completed after 3.5 minutes (Figure 2).
Carum Copticum extract in concentrations less
than 1% (0.001, 0.002 and 0.004) could not produce
Effect of Carum Copticum on acetylcholine induced contraction in rat’s ileum 77

30
Ach ingredients insert their action on tracheal smooth
CCE-Ach muscles via anticholinergic [21] antihistaminic [9]
25 and calcium channel blocking activity [6]. In the
Deviation from base line (mm)

present study the action of Carum Copticum extract

on rat’s ileal motility interfered with the contrac-
20
tile activity of acetylcholine on the specimens. The
inhibitory effect of Carum Copticum extract on
15 acetylcholine induced contraction seen in this study
may be due to β-adrenergic stimulatory action or
10 due to the inhibitory effect of this plant on cholin-
ergic receptors. The inhibitory effect of the extracts
of this plant were therefore, examined on isolated
5
ileum preparations pretreated with acetylcholine,
indicating a functional antagonistic effect of Carum
0 Copticum extract on cholinergic receptors in rat’s
1 ileum. The inhibitory action of Carum Copticum ex-
Figure 1 The effect of pre treating ileal specimens with tract on acetylcholine induced contraction was sim-
Carum Copticum extract (CCE) on acetylcholine (Ach) ilar but slower than that of atropine sulfate. The
induced contraction. major constituents of Carum Copticum extract me-
diating its relaxant effect are pinene components
25 [22−24], thymol [25,26] and carvacrol [27,28]. From
these components the anticholinergic activity of
Deviation from base line (mm)

thymol is reported [25], so the anticholinergic ef-

20
fect of Carum Copticum extract in this study may be
through the action of its thymol ingredient which
15 needs further investigations for more details.

10
Acknowledgments
5
The authors give thanks to all people who have
0 assisted the experimental procedure and to the
1 2 3 4 5 6 research deputy of Yazd Shahid Sadughi Medical
Time (min) University as the sponsor of this research.
Ach Ach + CCE Ach + Atr

Figure 2 The inhibitory effect of atropine (Atr) and References

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