Fitoterapia 77 (2006) 425 – 428 www.elsevier.com/locate/fitote Relaxant effect of aqueous extract of Cistus ladaniferus on rodent intestinal contractions Mohammed Aziz a,⁎, Naoual Tab a , Ahmed Karim a , Hassane Mekhfi a , Mohamed Bnouham a , Abderrahim Ziyyat a , Ahmed Melhaoui b , Abdelkhaleq Legssyer a a Laboratoire de Physiologie Ethnopharmacologie, UFR “Physiologie et Pharmacologie” Département de Biologie, Université Mohamed Premier, Faculté des Sciences, BP 717, 60000 Oujda, Morocco b Laboratoire de Phytochimie et Pharmacognosie, Département de Biologie, Université Mohamed Premier, Faculté des Sciences, BP 717, 60000 Oujda, Morocco Received 26 December 2005; accepted 12 May 2006 Available online 24 May 2006 Abstract The effects of the aqueous extract of Cistus ladaniferus leaves and stems were studied on the rodent isolated jejunum. The extract produced a reversible concentration dose-dependent (0.1–3 mg/ml) inhibition of the spontaneous motility of the rabbit jejunum. The inhibitory effects of the extract were not affected by pretreatment with the inhibitors of the α and β adrenergic receptors yohimbine, prazosin or propranolol. The extract also inhibited K+-induced contractions in rabbit and rat jejunum at a similar concentration range. This result suggests that the antispasmodic action of the extract is mediated through calcium channel blockade. The results confirm the traditional use of C. ladaniferus in treating intestinal ache. © 2006 Elsevier B.V. All rights reserved. Keywords: Cistus ladaniferus; Jejunum; Antispasmodic 1. Introduction In numerous countries medicinal plants were largely used by the population due to the low cost, easy access and ancestral experience . In these countries and in the last recent times numerous studies were realized to test the effect of the extracts of these plants on the intestinal contractions [2–6]. The present study was realized in order to valorize the plants of the region of our university with the aim to identify and evaluate commonly available natural drugs as alternative to currently used antispasmodic drugs which are not completely free from adverse effects . Cistus ladaniferus, a Cistaceae named rockrose, is present in the perimeter of Mediterranean sea. It is a medium sized tree locally known as “Touzal” in North-East of Morocco and/or “Targala and Bu-zgzaw” in the other regions of Morocco . It is present in the mountainous regions of the West Rif, central zone and the Beni-Znassen of Morocco. This plant is widely used in herbal medicine by the local population of the North-east of Morocco as anti-diarrhoeal, antiacid and ⁎ Corresponding author. Fax: +212 56 50 06 03. E-mail address: [email protected] (M. Aziz). 0367-326X/$ - see front matter © 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.fitote.2006.05.015 426 M. Aziz et al. / Fitoterapia 77 (2006) 425–428 Fig. 1. Effects of the C. ladaniferus (CL) aqueous extract on spontaneous contractions of isolated rabbit jejunum. ⁎P < 0.05, ⁎⁎P < 0.005 statistically significant difference from control. antispasmodic. Essential oil, absolute and resinoid of this plant had been shown to exhibit antifungal and antibacterial effect  and it is used in cosmetic industry [10–12]. Its aqueous extract exhibits an antiaggregant effect . The purpose of this study is to verify the antispasmodic effect of this plant on rodent isolated jejunum. 2. Experimental 2.1. Plant C. ladaniferus L.(Cistaceae), collected from Oujda region in the North-east of Morocco in April–May 2002 was identified by B. Haloui from the Biology Department of Oujda Sciences Faculty (Morocco). A voucher specimen (No rab 502/63) was previously deposited in Scientific Institute of Rabat. 2.2. Extraction C. ladaniferus leaves and stems (50 g) boiled in 1 l water and evaporated to dryness gave a crude residue (yield: 18%). Fig. 2. Effect of the C. ladaniferus (CL) aqueous extract on the carbachol (CCh)-induced contraction of isolated rat jejunum. M. Aziz et al. / Fitoterapia 77 (2006) 425–428 427 Fig. 3. A: Effect of noradrenaline (Nadr) on rabbit jejunum contractions. B: Effect of the pretreatment with adrenergic receptors antagonists (prazosin, yohimbine and propranolol) on the inhibitory effect of the aqueous extract of C. ladaniferus (CL) on rabbit jejunum contractions. 2.3. Animals Male Wistar rats weighing 250–350 g and New Zealand male rabbits (1.5–2 kg) were kept in a animal room and maintained in standard environmental conditions of temperature, humidity and light. Animals had free access to water and food until the experiment. Food was withdrawn 24 h prior to experiment. Fig. 4. Effect of the C. ladaniferus (CL) aqueous extract on K+-induced contraction of rabbit jejunum. 428 M. Aziz et al. / Fitoterapia 77 (2006) 425–428 2.4. Antispasmodic effect Animal was killed by cervical dislocation followed by exsanguinations. Segments of jejunum about 2–2.5 cm were suspended in oxygenated Tyrode's solution and maintained at 37 °C. It was allowed to equilibrate for at least 30 min before the addition of any drugs. The intestinal movement and response to stimulation were recorded. A concentration response was obtained by cumulative addition of aqueous extract. The effect of the extract against carbamylcholine chloride (carbachol, CCH), noradrenaline (NA) at 10− 6 M, yohimbine, prazosin, propranolol at 5 10− 5 M and KCl at 40 mM was evaluated. 2.5. Statistical analysis Student's t-test was used for statistical analysis and P < 0.05 was considered to be significant. 3. Results and discussion The jejunum of rabbit was used to study the spontaneous basal contractions because their amplitude was easier to evaluate than the rat jejunum. The aqueous extract of C. ladaniferus has exhibited a reversible inhibitory effect in a dose-dependent fashion (Fig. 1). The extract inhibited carbachol-induced contractions of jejunum of both rats and rabbit. In Fig. 2 we show only an effect on rat jejunum. This effect tends to suggest some anticholinergic activity likely mediated through cholinergic receptors. As envisaged we found that noradrenaline inhibits the rabbit intestinal basic contraction (Fig. 3A). The blocking of all adrenergic receptors, α1 by the prazosin and α2 by the yohimbine and β by the propranolol, shows that noradrenaline does not have any effect on the contractions; while the aqueous extract at the concentration of 3 mg/ml, the dose previously shown to produce a clear antispasmodic effect, causes the inhibition of these contractions (Fig. 3B). These results suggest that these effects were not mediated through the adrenergic receptors. The Ca++ plays an essential role in the cellular mechanisms regulating smooth muscle contraction . The contractions induced by high K+ depend on flux of Ca++ into the cells through voltage dependent Ca++ channels  and a substance, which inhibits K+ induced contractions, is considered as a calcium channel blocker. Thus, inhibition of high K+induced contractions of rabbit and rat jejunum by plant extract (Fig. 4) may be considered as an outcome of restricted Ca++ entry via voltage dependent Ca++ channels. These results tend to support the ethnomedical use of these plants for the treatment of gastrointestinal disorders. Acknowledgments Mustapha Badraoui and Karim Ramdaoui are acknowledged for technical support and animal breeding. This work was supported by grants from the Centre National de Recherche (CNR, Morocco), project PARS Médecine 081. References                Marin-Bettolo GB. 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