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Journal of Ethnopharmacology 134 (2011) 878–883

Contents lists available at ScienceDirect

Journal of Ethnopharmacology

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rokinetic and laxative activities of Lepidium sativum seed extract with speciesnd tissue selective gut stimulatory actions

ajeeb-ur-Rehmana,b, Malik Hassan Mehmooda,b, Khalid M. Alkharfyc, Anwarul-Hassan Gilania,c,∗

Natural Product Research Division, Department of Biological and Biomedical Sciences, The Aga Khan University Medical College, Karachi 74800, PakistanDepartment of Pharmacology, Faculty of Pharmacy, University of Karachi 75270, PakistanDepartment of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia

r t i c l e i n f o

rticle history:eceived 19 November 2010eceived in revised form 11 January 2011ccepted 27 January 2011vailable online 3 February 2011

eywords:epidium sativumruciferaerokineticaxative

a b s t r a c t

Aim of the study: To provide ethnopharmacological basis for the medicinal use of Lepidium sativum seedsin indigestion and constipation.Materials and methods: The in vivo studies were conducted in mice, while isolated tissues of mouse,guinea-pig and rabbit were suspended in tissue bath to measure isotonic contractions.Results: The aqueous-methanolic extract of Lepidium sativum seeds (Ls.Cr) at 30 and 100 mg/kg showedatropine-sensitive prokinetic and laxative activities in mice, which were partially sensitive to atropine.In isolated gut preparations of mouse and guinea-pig, Ls.Cr (0.1–1 mg/mL) caused a concentration-dependent stimulatory effects both in jejunum and ileum, which was blocked in the presence of atropine.In rabbit jejunum, the stimulant effect of Ls.Cr remained unchanged in the presence of atropine, pyril-amine or SB203186, while in rabbit ileum, the stimulatory effect was partially blocked by atropine. The

holinergicleumejunumpecies-specificodents

Ls.Cr was more efficacious in gut preparations of rabbit than in guinea-pig or mouse. The phytochemicalanalysis of the plant extract detected alkaloids, saponins and anthraquinones as plant constituents.Conclusion: This study showed the prokinetic and laxative effects of Lepidium sativum in mice, whichwere partially mediated through a cholinergic pathway. The in vitro spasmodic effect of the plant extractmediated through a similar mechanism with species and tissue-selectivity, provides a rationale for themedicinal use of the seeds of Lepidium sativum in indigestion and constipation, and suggests studying the

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plant extracts on more th

. Introduction

Lepidium sativum Linn. (family: Cruciferae) is commonly knowns “Common cress”, “Garden cress” or “Halim”. It is a small smoothrect annual herb and is cultivated as a culinary vegetable allver Asia (Baquar, 1989). Its seeds are popularly used as gastroin-estinal (GI) stimulant, laxative, gastroprotective and digestive aidNadkarni, 1986; Fleming, 1998). In addition, the plant has beeneported to have other properties, such as antibacterial, antiasth-atic, diuretic, aphrodisiac, and abortifacient; its leaves are also

sed by Europeans in salads (Nadkarni, 1986; Duke et al., 2002).

The plant has been reported to contain alkaloids (imidazole, lep-

dine, semilepidinoside A and B) (Maier et al., 1998), �-carotenes,scorbic, linoleic, oleic, palmitic and stearic acids (Duke, 1992),ucurbitacins and cardenolides (Fleming, 1998). Moreover, a few

∗ Corresponding author at: Department of Biological and Biomedical Sciences, Thega Khan University Medical College, Karachi 74800, Pakistan. Tel.: +92 21 4864571;

ax: +92 21 4934294/2095.E-mail address: anwar.gilani@aku.edu (A.-H. Gilani).

378-8741/$ – see front matter © 2011 Elsevier Ireland Ltd. All rights reserved.oi:10.1016/j.jep.2011.01.047

e species to get the wider picture.© 2011 Elsevier Ireland Ltd. All rights reserved.

phenolic constituents, such as, sinapic acid and sinapin were iso-lated from its seed extract (Schultz and Gmelin, 1952).

Lepidium sativum has been studied for its medicinal use asantihypertensive (Maghrani et al., 2005), diuretic (Patel et al.,2009), antiasthmatic (Archana and Anita, 2006), antiinflammatory,hypothermic, analgesic and coagulant (Al-Yahya et al., 1994), hypo-glycemic (Patole, 1998) and bone fracture healing agent (Ahsanet al., 1989). However, there is no report in the literature validat-ing its medicinal use in constipation or other GI disorders. In thisinvestigation, we studied the seed extract of Lepidium sativum usingthe in vivo and in vitro assays, to provide a scientific base for itsmedicinal use in gut disorders, such as indigestion and constipation.

2. Material and methods

2.1. Preparation of the crude extract

The seeds of Lepidium sativum were purchased from a herbalstore (Bin Menqash, Riyadh, Saudi Arabia) in March, 2010. Theplant was authenticated by Dr. Mohammed Yusuf of King SaudUniversity and the specimen has been preserved at the herbarium

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f the College of Pharmacy, King Saud University, Riyadh, Saudirabia, and also at the Natural Product Research Division, Depart-ent of Biological and Biomedical Sciences, Aga Khan University,

arachi with voucher # Ls-SE-04-10-98. By following a previouslyescribed method (Williamson et al., 1998), the seeds of Ls.Cr wereoaked in 70% methanol for 3 d and filtered through muslin clothnd Whatman (Maidstone, UK) No. 1 filter paper simultaneously.his procedure was repeated three times, and all the filtrates wereooled and evaporated in rotary evaporator (model RE-111, Buchi,lawil, Switzerland) under a reduced pressure to obtain finally, therude extract of the seeds of Lepidium sativum (Ls.Cr). The yield ofhick dark brown pasty like mass was 15% (wt/wt).

.2. Phytochemical screening

Phytochemical analysis of the crude extract of Lepidium sativumas performed qualitatively for the presence of alkaloids, saponins

nd anthraquinones as plant constituents according to a standardethod (Evans, 2006). Briefly, alkaloids were detected by treat-

ng the plant material with Dragendor’s reagent, resulting in theppearance of a precipitate at the bottom of the test tube. Theresence of saponins was based in the appearance of froth uponigorous shaking of diluted samples. Lastly, anthraquinones werecreened, by treatment of the plant material with NH4OH reagentdded after dissolving it in 1% HCl and purifying with benzene.he appearance of a pink, violet, or red color demonstrated theirresence.

.3. Drugs

Acetylcholine perchlorate (ACh), atropine sulphate, carbamyl-holine (CCh), histamine hydrochloride, 5-hydroxytryptamine5-HT), pyrilamine maleate and hexamethonium chloride wereurchased from Sigma–Aldrich Chemicals Company (St Louis, MO,SA). SB203186 (1-piperidinylethyl-1H-indole-3-carboxylate) wasurchased from Tocris (Ballwin, MO, USA). Chemicals used foraking physiological salt solutions including potassium chloride,

alcium chloride, glucose, magnesium chloride, magnesium sulfate,otassium dihydrogen phosphate, sodium bicarbonate, sodiumihydrogen phosphate and sodium chloride were obtained fromerck (Darmstadt, Germany). All chemicals used were of the ana-

ytical grade available and solubilized in distilled water.

.4. Animals

A total of 90 BALB/c mice (weighing 20–25 g), 5 guinea-pigsweighing 400–600 g) and 7 local breed rabbits (weighing 1–1.5 kg)f either sex were used, which were housed at the animal housef The Aga Khan University under a controlled environment23–25 ◦C). The animals were kept in plastic cages (47 × 34 × 18)ith sawdust (changed at every 48 h) and were fasted for 24 h

efore starting the experiment. In routine, they were given tapater ad libitum and a standard diet (Harkness and Wagner, 1995)

onsisting of (g/kg): flour 380, fiber 380, molasses 12, NaCl 5.8,utrivet L 2.5, potassium metabisulfate 1.2, vegetable oil 38, fisheal 170 and powdered milk 150. The experiments were per-

ormed with the rulings of the Institute of Laboratory Animalesources, Commission on Life Sciences, National Research Coun-il (National Research Council, 1996) and approved by the Ethicsommittee of The Aga Khan University.

.5. In vivo experiments

.5.1. Charcoal meal GI transit testThe method described by Al-Qarawi et al. (2003) was followed

ith slight modifications. Mice fasted for 12 h were divided into 7

pharmacology 134 (2011) 878–883 879

different groups (6 animals in each). Two of the groups were treatedper oral (p.o.) with increasing doses of Ls.Cr 30 and 100 mg/kg, act-ing as the test groups. One group was taken as negative control,treated with saline (10 mL/kg). The next group was administeredCCh (1 mg/kg) as the positive control. After 15 min, the animalswere given 0.3 mL of charcoal meal of distilled water suspensioncontaining 10% gum acacia, 10% vegetable charcoal and 20% starch.The animals were sacrificed after 30 min and the abdomen wasopened to excise the whole small intestine. The length of the smallintestine and the distance between the pylorus region and the frontof the charcoal meal was measured to obtain the charcoal transportratio or percentage. In order to assess the involvement of acetyl-choline (ACh)-like prokinetic effect of the extract and CCh, furthergroups of mice were pretreated with intraperitonial (i.p.) injec-tion of atropine (10 mg/kg) 15 min prior the administration of theextract or CCh.

2.5.2. Laxative activity testIn accordance with the previous method (Haruna, 1997), mice

fasted for 6 h before the experiment were placed individually incages lined with clean filter paper. The animals were divided into7 groups (6 animals in each); the first group acting as the nega-tive control and administered saline (10 mL/kg, p.o.), while the nextgroup received CCh (1 mg/kg, i.p.), which served as the positive con-trol. The third and fourth groups received orally, 30 and 100 mg/kgof Ls.Cr, respectively. To determine the mechanism underlyingits laxative effect, separate sets of mice (group # 5–7) were pre-treated with atropine (10 mg/kg, i.p.) 1 h before administration ofthe extract or CCh. After 18 h, the feces production (total number offeces and total number of wet feces per group) in all animals wascounted, and the percentage increase in wet feces relative to thatof total fecal output was recorded, which was considered as thelaxative effect (Mehmood and Gilani, 2010).

2.6. In vitro experiments

By following the previously described methods (Ghayur andGilani, 2005; Ghayur et al., 2005; Gilani et al., 2005), differentgut preparations (jejunum and ileum) from mouse, guinea-pig andrabbit were obtained subsequent to cervical dislocation of the ani-mals; the abdomen was cut opened, required tissues were isolatedout. Tissue preparations (ileum or jejunum) of 2–3 cm long weremounted in 10 mL tissue baths containing Tyrode’s solution main-tained at 37 ◦C and aerated with a mixture of 5% carbon dioxide and95% oxygen (carbogen). The composition of Tyrode’s solution (mM)was KCl 2.68, NaCl 136.9, MgCl2 1.05, NaHCO3 11.90, NaH2PO4 0.42,CaCl2 1.8, and glucose 5.55 (pH 7.4). A preload of 1 g was applied toeach tissue, and the contractile responses were recorded using iso-tonic transducer 50-6360 (Harvard Apparatus, Holliston, MA, USA)coupled with either a student oscillograph (Harvard Apparatus) orPowerLab (ML-845) data acquisition system (AD Instruments; Syd-ney, Australia) and a computer using chart software (version 5.3).The tissues were allowed to equilibrate for a period of 30 min, andthen stabilized with sub-maximal concentration of acetylcholine(ACh, 0.3 �M). The tissues were presumed stable only after thereproducibility of the said responses. The Lepidium sativum seedextract was examined later for any spasmodic activity on the ileumand jejunum preparations of mouse, guinea-pig and rabbit at con-centrations ranged from 0.003 to 3.0 mg/mL.

2.7. Statistical analysis

All the data expressed are mean ± standard error of mean(S.E.M., n = number of experiments) and the median effectiveconcentrations (EC50 values) with 95% confidence intervals (CI).One-way analysis of variance (ANOVA) followed by Dunnett’s test

8 Ethnopharmacology 134 (2011) 878–883

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80 Najeeb-ur-Rehman et al. / Journal of

r unpaired t-test was used to assess the laxative activity, whilene-way ANOVA followed by Tukey’s test was employed for theffect of plant extract in charcoal meal transit. The concentration-esponse curves (CRCs) were analyzed by non-linear regression andwo-way ANOVA followed by Bonferroni’s post-test correction ornpaired t-test was used for multiple comparisons of CRCs withhe respective control. All the graphing, calculations and statisti-al analysis were performed using GraphPad Prism 4 for windowsGraphPad Software, San Diego, California, USA).

. Results

.1. Phytochemical analysis

Preliminary phytochemical analysis of the seed extract of Lep-dium sativum revealed the presence of alkaloids, saponins andnthraquinones.

.2. In vivo findings

.2.1. Effect of Ls.Cr on charcoal mealLepidium sativum seed extract dose-dependently

30–100 mg/kg) propelled charcoal meal through the smallntestine of mice (Fig. 1). The distance travelled by the salinereated group was 57.1 ± 2.6% (mean ± S.E.M., n = 6) of totalength of small intestine, while the positive control group ofCh (1 mg/kg) significantly enhanced the movement (p < 0.001s. saline) of charcoal meal to 96.3 ± 2.9%. The plant extract athe dose of 30 and 100 mg/kg moved charcoal meal to the levelf 73.9 ± 1.8% (p < 0.01) and 86.7 ± 2.8% (p < 0.001), respectively,hen compared with the saline treated group. When the plant

xtract (30 and 100 mg/kg) and positive control (CCh, 1 mg/kg)roups were restudied for their influence on transit of charcoaleal, in mice pretreated with atropine, all the excitatory effectsere markedly decreased as seen in Fig. 1.

.2.2. Laxative activityLs.Cr treatment produced 53.8 ± 9.2% and 63.2 ± 5.3% (n = 6) wet

eces in mice at 30 and 100 mg/kg, respectively. The positive con-rol, CCh (1 mg/kg) produced 73.6 ± 6.8% wet feces, while the salinereated group did not form any wet feces. When Ls.Cr (30 and00 mg/kg) was studied for its positive influence on wet feces inice pretreated with atropine, the effect declined to 4 ± 4% and

.8 ± 5%, respectively; further details are shown in Table 1.

.3. In vitro findings

.3.1. Effect of Ls.Cr on jejunum preparation of different animalsIn mouse jejunum, the seed extract of Lepidium sativum caused

concentration-dependent stimulant effect at 0.03–0.3 mg/mL,eing more effective than in mouse ileum. Pretreatment of theissue with atropine (0.1 �M) blocked (p < 0.001) the spasmogenic

able 1ffect of atropine on the laxative activity of the seed extract of Lepidium sativum (Ls.Cr) in

Group no. Treatment Dose (mg/kg) Mean defe

1 Saline (p.o., mL/kg) 10 1.4 ± 0.2 Carbachol (i.p.) 1 11.4 ± 0.3 Ls.Cr (p.o.) 30 6.8 ± 1.4 100 9.4 ± 1.5 Carbachol (i.p.) + Atropine (i.p.) 1 + 10 1.9 ± 0.6 Ls.Cr (p.o.) + Atropine (i.p.) 30 + 10 3.6 ± 0.7 100 + 10 4.8 ± 0.

alues shown are mean ± S.E.M. of 6 animals per group. *p < 0.05, **p < 0.01 and ***p < 0.00y Dunnett’s test), group #5 vs. group #2, group #6 vs. group #3 and group #7 vs. group #

njection.

the absence and presence of atropine. *p < 0.05, **p < 0.01 and ***p < 0.001, one-wayANOVA followed by Tukey’s test. Each bar shown represents mean ± S.E.M. of 6animals per group.

effect of Ls.Cr, while the presence of hexamethonium, pyrilamineor SB203186 did not alter (p > 0.05) its effect (Fig. 2A). The val-ues shown in Fig. 2A represent 4–6 isolated tissue preparationsobtained from 3 animals.

In guinea-pig jejunum, Ls.Cr (0.01–0.1 mg/mL) exhibited a mildspasmodic effect reaching its highest 17.4 ± 1.2% (mean ± S.E.M.;n = 5) of ACh maximum. The efficacy of achieved contractile effectof the extract in jejunum preparation was found lower thanobserved in guinea-pig ileum. When the spasmogenic effect ofLs.Cr was redetermined in the presence of different antagonists,it was fully blocked in the presence of atropine, while remainedunchanged (p > 0.05) in the presence of hexamethonium, pyril-amine or SB203186 (Fig. 2B). The values presented in Fig. 2Brepresent 4–5 isolated tissue preparations obtained from 2 animals.

When tested in spontaneously contracting rabbit jejunum, theplant exhibited a concentration-dependent (0.1–1 mg/mL) stimu-latory effect. The efficacy of the spasmogenic effect was 66.6 ± 2.7%(n = 7) of ACh maximum, observed at 1 mg/mL. The efficacy of Ls.Cr

was less in rabbit jejunum than in its ileum preparations (p < 0.001).The spasmodic effect of the plant remained unchanged (p > 0.05)when restudied in tissues pretreated with atropine, hexametho-nium, pyrilamine or SB203186, as seen in Fig. 2C. The values shown

mice.

cation/group Mean number of wet feces/group Mean % of wet feces

6 0 06** 8.4 ± 0.9** 73.6 ± 6.81* 3 ± 0.4** 53.8 ± 9.28** 4.8 ± 0.4** 63.2 ± 5.38*** 0.1 ± 0.09*** 4.0 ± 1.75* 0.2 ± 0.2** 4 ± 47** 0.4 ± 0.2*** 7.8 ± 5

1 show a comparison of group #2, 3 and 4 vs. group #1 (One-way ANOVA followed4 (unpaired t-test). The term p.o. represents per oral, while i.p. is for intraperitoneal

Najeeb-ur-Rehman et al. / Journal of Ethnopharmacology 134 (2011) 878–883 881

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ig. 2. The stimulatory effects of the seed extract of Lepidium sativum (Ls.Cr) without1 �M) in isolated jejunum and ileum preparations of (A and D) mouse, (B and E)xperiments conducted on the isolated tissue preparations from 5 to 7 differenttwo-way ANOVA, followed by Bonferroni’s post-test correction or unpaired t-test)

n Fig. 2C represent 4–7 isolated tissue preparations obtained from

animals.

.3.2. Effect of Ls.Cr on ileum preparation of different animalsIn mouse ileum, the seed extract of Lepidium sativum caused a

timulatory effect at 0.03–1 mg/mL in isolated mouse ileum with

ith atropine (0.1 �M), pyrilamine (1 �M), hexamethonium (0.3 mM) and SB203186a-pig and (C and F) rabbit. The values shown are mean ± S.E.M. of 4–7 individualls [mouse (6), guinea-pig (5) and rabbits (7)]. *p < 0.05, **p < 0.01 and ***p < 0.001

efficacy of 22.3 ± 1.1% (n = 5) of the ACh maximum obtained at

3 �M (Fig. 2D). Pretreatment of the tissue with atropine (0.1 �M)blocked the spasmogenic effect of Ls.Cr, while hexamethonium(0.3 mM), pyrilamine (1 �M) or SB203186 (1 �M) pretreatmentshowed no influence (Fig. 2D). The values presented in Fig. 2D aremean ± S.E.M. of 4–6 preparations obtained from 3 animals.

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In guinea-pig ileum, the plant extract exhibited concentration-ependent (0.01–1 mg/mL) contractions with efficacy of5.8 ± 2.7% (n = 6) of ACh maximum. Pretreatment of the tissueith atropine (0.1 �M), but not with hexamethonium, pyrilamine

r SB203186, abolished the spasmogenic effect of Ls.Cr (Fig. 2E).he values shown in Fig. 2E are mean ± S.E.M. of 4–6 preparationsbtained from 3 animals.

In rabbit ileum, Ls.Cr also showed a concentration-dependent0.1–3 mg/mL) stimulatory effect with efficacy (98.8 ± 0.9%) simi-ar to that ACh (p > 0.05). Pretreatment of the tissue with atropine0.1 �M) partially blocked the spasmogenic effect at higheroncentration (p < 0.001), while pretreatment of tissues with hex-methonium, pyrilamine or SB203186 had no effect (p > 0.05) ashown in Fig. 2F. The values presented in Fig. 2F are mean ± S.E.M.f 5–7 preparations obtained from 4 animals.

. Discussion

Keeping in view the medicinal use of Lepidium sativum in gutisorders, such as indigestion and constipation, its seed extract wasested in mice, where it propelled charcoal meal through the smallntestine and increased the production of wet feces, hence showingrokinetic and laxative activities, similar to the effect of carbachol,standard cholinergic agonist and accelerator of intestinal con-

ents (Brown and Taylor, 2006). These gut stimulatory actions ofhe extract were found partially sensitive to atropine, a muscariniceceptor blocker (Gilani et al., 1997), indicating the presence ofome ACh-like component(s) in addition to other gut stimulantonstituent(s). ACh is a neurotransmitter of the parasympatheticervous system and is known to cause gastrointestinal stimulationhrough the activation of muscarinic receptors (Brown and Taylor,006), hence, the presence of ACh-like constituents explains itsedicinal use in constipation and as digestive aid.To further study the possible mode of the observed prokinetic

nd laxative properties of the extract, we used isolated jejunum andleum preparations from different animals, such as mouse, guinea-ig and rabbit. In the gut preparations from mouse and guinea-pig,hough, efficacy for the spasmodic effect of the plant extract var-ed, but the stimulant effect was completely blocked by atropine inoth preparations, thus, showing a common mechanism, throughholinergic action. However, when tested in the gut preparations ofabbit, the stimulant effect in jejunum was not blocked by any of thentagonists studied including pyrilamine, a histaminic type-1 (H1)eceptor blocker (Sharif et al., 1994) or hexamethonium, a ganglionlocker (Wien et al., 1952) or SB203186, a serotonergic receptorntagonist (Sander-Bush and Mayer, 2006), while atropine causedpartial blockade in ileum, but no effect on jejunum. This indicates

hat the rabbit tissue behaves differently from those of mouse anduinea-pig in the nature of gut stimulant effect, and clearly sug-esting some additional mechanism(s), independent of histamine,icotine or 5-hydroxytryptamine (5-HT, serotonin) receptors acti-ation.

Taken together, these results demonstrate the presence oftropine-sensitive stimulatory constituent(s) in Lepidium sativumbserved in all tissues except rabbit jejunum, and atropine-nsensitive observed in both tissues of rabbit. The spasmogenicffect of Ls.Cr on rabbit tissues seems involving more than onetimulant components, hence, showed the highest efficacy. Otherechanisms known for their gut stimulant property, which have

ot been ruled out in this study includes certain prostaglandins

Beubler and Kollar, 1988), platelet activating factor (Izzo et al.,998), nitric-oxide-donating or releasing compounds (Mascolot al., 1994), dopaminergic antagonists (Briejer et al., 1995), chole-ystokinin (Briejer et al., 1999) and tachykinins (Severini et al.,002).

pharmacology 134 (2011) 878–883

Collectively, the data in jejunum and ileum preparations ofmouse, guinea-pig and rabbit indicate a species and tissue-selectivegut stimulatory effect of Lepidium sativum extract, which was evi-dent in the following ways: (1) in rabbit jejunum preparations,the stimulatory effect of the crude extract was insensitive toatropine and was greater (p < 0.01) than the achieved effect of Ls.Cron all other tissues, showing partially or fully atropine sensitiveeffect and (2) in ileum preparations, the stimulatory effect of theextract was partially sensitive to atropine and was more efficacious(p < 0.001) in rabbit as compared to its observed atropine-sensitiveeffect in guinea-pig and mouse. In different used animals, theefficacy of Ls.Cr for its stimulant effect was as rabbit > guinea-pig > mouse.

In imparting its spasmogenic effect, Ls.Cr was found more effi-cacious in rabbit and guinea-pig ileum, while in mouse, it was moreeffective in jejunum. Such types of tissue or species-selective effectshave also been noticed in various earlier studies with other plantextracts (McLeod et al., 1994; Ghayur and Gilani, 2005; Ghayuret al., 2005; Mehmood and Gilani, 2010; Mehmood et al., 2010).Based on these observations, a suggestion can be made that whenstudying the gut stimulatory effect of the plant materials using guttissues from more than one species may be advisable to know thebroader picture.

The presence of saponins and anthraquinones amongst thechemical constituents, which are known for their spasmogeniceffect (Akah et al., 1997; Pasricha, 2006), may explain the gut stim-ulant actions of Lepidium sativum.

5. Conclusion

This study shows that the prokinetic and laxative activities ofLepidium sativum in mice are partially mediated through mus-carinic receptors, and the in vitro findings demonstrated the speciesand tissue-selective spasmodic effect mediated through a similarmechanism. Thus, this study provides sound mechanistic basis forthe medicinal use of Lepidium sativum in gut disorders, such asindigestion and constipation, and the observed specifies and tissue-selective effect in this study suggest using more than one speciesfor a better picture of the pharmacological profile when studyingplant extracts.

Acknowledgements

This study was initiated during the visit of Prof. Anwarul-Hassan Gilani to King Saud University as a part of Visiting ProfessorProgramme, and partially supported by the Higher EducationCommission, Government of Pakistan, as an indigenous Ph.D. schol-arship to Mr. Najeeb-ur-Rehman.

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