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 S.H. Hejazian-Y et al

expectorant, diuretic, anti-vomiting, analgesic, anti- Preparations were suspended from the transducer
asthma, anti-dyspnea, and anti-spasm, is postulated lever in the axis of the longitudinal muscle with
for Carum Copticum fruits [2]. Despite the availa- fine thread, mounted in the 50 mL internal chamber
bility of modern medications the propensity towards of organ bath containing Tyrode’s solution, main-
the traditional medications is growing throughout tained at 37ºC and bubbled with 95% O2 and 5% CO2
the world [3] which needs scientific investigations [13]. Then, it was allowed to stabilize for 60 min-
for evaluating the therapeutic effects of medicinal utes prior to drug addition. Isotonic responses were
plants and their mechanisms of actions. Some of recorded using an isotonic transducer (T2) and an
these biological investigations have been focused oscillograph recording system (the bioscience 400
on gastrointestinal dysfunctions. One of the most Series Washington oscillograph). Acetylcholine (Sigma
important abnormalities in gastrointestinal tract is Aldrich Chemie Gmbh, Germany) was used as a stan-
diarrhea, which may be produced by some infec- dard contractile control at the end of each assay.
tions or increase in motility [4,5]. Carum Copticum After recording a base line as a control for ideal
is traditionally used in treating many gastrointesti- mechanical activity, the specimens were influenced
nal disorders such as indigestion, colic, dyspepsia by different concentrations of Carum Copticum ex-
and diarrhea [6] and is suggested to be anti nocic- tract in a cumulative manner to yield the minimum
eptive [7] as well. It has been found that Carum effective concentration which was 1% of total ex-
Copticum is very effective against some bacteria tract diluted in Tyrode’s solution. The effects of 1%
and its effect on digestive tract may be due to its Carum Copticum extract alone, after and before
antibacterial activity [8]. In other hand there are the infusion of acetylcholine (5 × 10−4 M) was assessed
evidences indicating the relaxant effects of Carum on the specimens.
Copticum on smooth muscles [9,10] and in our pre- The specimens were also influenced by 5 × 10−6 M
vious study we found that Carum Copticum extract atropine sulfate (Sigma Aldrich Chemie Gmbh) fol-
reduces the motility of rat ileum [11]. Intestinal lowing the acetylcholine infusion as a standard
motility is controlled by circulatory hormones, in- cholinergic antagonist. For each drug assay five sets
trinsic and extrinsic nerves that release a variety of experiments were done.
of transmitters. The most important excitatory The data were calculated as mean changes in
transmitter in gastrointestinal tract is acetylcho- specimen length from base line due to changes in
line which acts via muscarinic receptors in enteric contractile activity and analyzed by paired Student’s
smooth muscle cell membranes [12]. This study t-test. The statistical significance was considered
was conducted to understand the probable anti- as p ≤ 0.05.
cholinergic effect of Carum Copticum fruit extract
on isolated rat’s ileum.
3. Results

2. Materials and Methods Our findings showed that the Carum Copticum ex-
tract itself declines the amplitude of base line con-
In this study male Albino rats living in the standard tractions, in all five sets of experiments (3 ± 0.57 mm
environmental and feeding conditions were used vs. 4.28 ± 1.11 mm) which this relaxant effect was
for isolation of ileum. For experimental procedure significant (p = 0.022). The effective dose of acetyl-
permission of the animal ethics committee of Shahid choline conversely significantly (p = 0.000) elevated
Sadughi Medical University (Yazd, Iran), in accord- the amplitude of base line contractions in all as-
ance with the internationally accepted principles sessments (19.57 ± 4.6 mm vs. 4.28 ± 1.11 mm). After
for laboratory animal use and care mentioned by the pretreating the specimens with 1% Carum Copticum
European Community guidelines were obtained. extract, the effective concentrations of acetylcho-
For extraction preparation, 40 g air-dried fruits line only led to about 40% of its maximum response
of Carum Copticum (identified by botanists in the (Figure 1).
herbarium of Yazd herbal drug research center and The contractile effects of acetylcholine were
a sample specimen was stored and coded in this diminished by the infusion of atropine sulfate or
center) were gently grounded and mixed with 250 mL Carum Copticum extract.
of double distilled water (DDW) and subjected to The inhibitory effect of atropine sulfate (10−4 M)
steam distillation for 3 hours at atmospheric pressure. on acetylcholine induced contraction was observed
Experiments were performed as described in after a few second and completed in 30 seconds,
previous reports [13,14]. Adult male rats weighing while the inhibition due to 1% Carum Copticum ex-
200−250 g were sacrificed by cervical dislocation. tract was completed after 3.5 minutes (Figure 2).
Segments of ileum (2 cm in length), were obtained, Carum Copticum extract in concentrations less
flushed of their contents and trimmed of mesentery. 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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