Journal of Acupuncture and Meridian Studies
Volume 2, Issue 1 , Pages 75-78, March 2009

The Effect of Carum Copticum Extract on Acetylcholine Induced Contraction in Isolated Rat's Ileum

  • Seyed Hassan Hejazian-Y

      Affiliations

    • Herbal Medicine Research Center, School of Medicine, Shahid Sadughi University of Medical Sciences and Health Services, Yazd, Iran
    • ,
  • Mohammad Hossein Dashti-R

      Affiliations

    • Department of Physiology, School of Medicine, Shahid Sadughi University of Medical Sciences and Health Services, Yazd, Iran
    • Corresponding Author InformationCorresponding author. Department of Physiology, Shahid Sadughi School of Medicine, Booali St. Safaeyah, Yazd 8944157963, Iran
    • ,
  • Seyed Majid Mahdavi

      Affiliations

    • Department of Pharmacology, School of Medicine, Shahid Sadughi University of Medical Sciences and Health Services, Yazd, Iran
    • ,
  • Masood Anvar Qureshi

      Affiliations

    • Department of Physiology, Dow Medical College, Dow University of Health Sciences, Karachi, Pakistan

Received 5 August 2008; accepted 13 January 2009.

Article Outline

Abstract 

Aims

There are many biological investigations for determining an effective cure for the dysfunction of gastrointestinal tracts, using herbal medicine. It has been reported that Carum Copticum is a bactericidal agent and possesses anticholinergic, antihistaminic and b-adrenergic stimulatory effects in some tissues. However, these 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 effects of Carum Copticum on mechanical activity of isolated rat's 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 oscillographic 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.

Key Words:  acetylcholine , Carum Copticum , contraction , ileum

 

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1. Introduction 

Carum Copticum (L.) is a plant in Umbelliferae family with a white flower and small brownish seeds. This plant commonly grows in Iran, India, Egypt and Europe. The major component of its fruit is essential oil which is composed of g-terpinene, p-cymene, a-pinene, b-pinene, and other substances such as thymol and carvacol [1]. In the Iranian traditional medicine several therapeutic effects including expectorant, diuretic, anti-vomiting, analgesic, anti-asthma, anti-dyspnea, and anti-spasm, is postulated for Carum Copticum fruits [2]. Despite the availability 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 infections or increase in motility [4, 5]. Carum Copticum is traditionally used in treating many gastrointestinal disorders such as indigestion, colic, dyspepsia and diarrhea [6] and is suggested to be anti nociceptive [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 previous study we found that Carum Copticum extract reduces the motility of rat ileum [11]. Intestinal motility is controlled by circulatory hormones, intrinsic and extrinsic nerves that release a variety of transmitters. The most important excitatory transmitter in gastrointestinal tract is acetylcholine 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.

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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 accordance 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. 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, maintained at 37°C and bubbled with 95% O2 and 5% CO2[13]. Then, it was allowed to stabilize for 60 minutes 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 standard 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 extract in a cumulative manner to yield the minimum effective concentration which was 1% of total extract 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) following 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.

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3. Results 

Our findings showed that the Carum Copticum extract itself declines the amplitude of base line contractions, 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 acetylcholine conversely significantly (p = 0.000) elevated the amplitude of base line contractions in all assessments (19.57 ± 4.6 mm vs. 4.28 ± 1.11 mm). After pretreating the specimens with 1% Carum Copticum extract, the effective concentrations of acetylcholine 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 extract 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 an inhibitory effect on acetylcholine induced contractions.

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4. Discussion 

According to our findings Carum Copticum extract induced a relaxant effect on ileal smooth muscle contraction induced by acetylcholine. Different mechanisms are involved in gastrointestinal smooth muscle relaxation. These include the blocking action on excitatory agents and inducing anticholinergic [15] or antihistaminic [16] activity or act via their agonistic action on inhibitory modulators such as adrenergic [17], purinergic [18], GABAergic [19] agents or due to their nitric oxide mimetic action [20]. There are some evidences that Carum Copticum ingredients insert their action on tracheal smooth muscles via anticholinergic [21] antihistaminic [9] and calcium channel blocking activity [6]. In the present study the action of Carum Copticum extract on rat's ileal motility interfered with the contractile activity of acetylcholine on the specimens. The inhibitory effect of Carum Copticum extract on acetylcholine induced contraction seen in this study may be due to b-adrenergic stimulatory action or due to the inhibitory effect of this plant on cholinergic receptors. The inhibitory effect of the extracts of this plant were therefore, examined on isolated ileum preparations pretreated with acetylcholine, indicating a functional antagonistic effect of Carum Copticum extract on cholinergic receptors in rat's ileum. The inhibitory action of Carum Copticum extract on acetylcholine induced contraction was similar but slower than that of atropine sulfate. The major constituents of Carum Copticum extract mediating its relaxant effect are pinene components [22, 23, 24], thymol [25, 26] and carvacrol [27, 28]. From these components the anticholinergic activity of thymol is reported [25], so the anticholinergic effect of Carum Copticum extract in this study may be through the action of its thymol ingredient which needs further investigations for more details.

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Acknowledgments 

The authors give thanks to all people who have assisted the experimental procedure and to the research deputy of Yazd Shahid Sadughi Medical University as the sponsor of this research.

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References 

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PII: S2005-2901(09)60019-4

doi:10.1016/S2005-2901(09)60019-4

Journal of Acupuncture and Meridian Studies
Volume 2, Issue 1 , Pages 75-78, March 2009

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