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Journal of Ethnopharmacology 128 (2010) 15–19

Contents lists available at ScienceDirect

Journal of Ethnopharmacology

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creening of analgesic and anti-inflammatory activities of Citrullus colocynthisrom southern Tunisia

elsem Marzouka, Zohra Marzoukb, Ehsen Halouib, Nadia Feninab,bderrahman Bouraouic, Mahjoub Aounia,∗

Laboratoire des Maladies Transmissibles et Substances Biologiquement Actives, Faculté de Pharmacie, Monastir, Rue AVICENNE, 5000 Monastir, TunisiaUnité de Pharmaco-économie et Développement des Médicaments, Laboratoires de Biologie Végétale et Laboratoire de Pharmacologie, Faculté de Pharmacie, Monastir, TunisiaUnité URSAM, Laboratoire de Pharmacologie, Faculté de Pharmacie, 5000 Monastir, Tunisia

r t i c l e i n f o

rticle history:eceived 24 June 2009eceived in revised form 3 November 2009ccepted 26 November 2009vailable online 3 December 2009

eywords:itrullus colocynthis Schrad.queous extractcute toxicitynalgesic activitynti-inflammatory activityunisian medicinal plant

a b s t r a c t

Ethnopharmacological relevance: Inflammations and immune-related diseases such as rheumatoid arthri-tis are growing global concerns. Most of the drugs from plants which have become important in modernmedicine had a folklore origin and are traditional in systems of medicine. Citrullus colocynthis Schrad.(cucurbitaceae), endemic in Southern Tunisia, is used in folk medicine to treat many inflammationdiseases.Aim of study: To evaluate the acute toxicity of different parts of Citrullus colocynthis and then to screenthe analgesic and anti-inflammatory activities of aqueous extracts from roots and stems of the plant andfrom fruits and seeds at different maturation stages.Materials and methods: After identification and acute toxicity assay Citrullus colocynthis Schrad. aqueousextracts were screened for analgesic and anti-inflammatory activities using, respectively, the acetic acidwrithing test in mice and the carrageenan-induced paw edema assay in rats.

Results: All extracts displayed analgesic and anti-inflammatory activities at different doses without induc-ing acute toxicity. Topic results were obtained with immature fruits followed by seeds. The stem androot extracts were shown to possess the less significant inhibitory activity against analgesic and anti-inflammatory models.Conclusions: Based on this study, we confirmed that Citrullus colocynthis Schrad. is a potentially usefuldrug suitable for further evaluation for rheumatoid arthritis, and its folk medicinal use as an analgesicand anti-inflammatory agents is validated.

. Introduction

For the most part, modern science has neglected botanicalource of knowledge and the pharmacological potential of oldemedies have not been systematically evaluated. Neverthelessany species used in traditional medicine have the potential

o provide pharmacologically active natural products. Citrullusolocynthis Schrad. is a wild native plant growing in arid areasPottier-Alapetite, 1981). This cucurbitaceae is widely used inunisian folk medicine and it possesses therapeutic activities

gainst a wide range of ailments including inflammatory disorders,rthritis and gout (Le Flock, 1983; Boukef, 1986; Marzouk et al.,009). Nevertheless, a human overdose of plant immature fruits isazard. Intoxication is manifested by colitis, gastro-intestinal irri-

∗ Corresponding author. Tel.: +216 73 461 000; fax: +216 73 461 830.E-mail address: mahjoub.ouni@fphm.rnu.tn (M. Aouni).

378-8741/$ – see front matter © 2009 Elsevier Ireland Ltd. All rights reserved.oi:10.1016/j.jep.2009.11.027

© 2009 Elsevier Ireland Ltd. All rights reserved.

tations, delirium, hypothermia and cerebral congestions (Meyer,1989; Bellakhdhar, 1999).

In Tunisia, as in other Mediterranean countries (Al-Rawi andChaakravarty, 1964), the parts of plants most often used for medic-inal purposes are fruits and/or seeds, though other parts of theplants can be used, for example roots to treat urinary infection(Nadkami, 1954) or leaves (Batanouny, 1999). Traditional heal-ers seem to not pay attention to the plant’s degree of maturity.The literature rarely mentions if seeds are present in preparationsinvolving ground fruit/pulp. Modes of preparation and administra-tion vary, even for similar indications. Common preparations usefresh, warmed or dried plant material (often ground), as well asextracts used mostly in a liquid form. Extracts are prepared either

in water or in aqueous mixtures containing more lipophilic com-pounds (hot milk extractions, water/olive oil at various ratios) ata temperature ranged from tepid to boiling. Ground plant materialcan be mixed with honey for ingestion or topical gynaecologicalapplication or with other plants for poultices (for example with

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awsonia inermis and Capparis spinosa). Methods of administrationre topical, rectal or vaginal (fruit), enema, cervico-vaginal douche,nd by ingestion (Boukef, 1986; Marzouk et al., 2009). Extreme cau-ion should be exercised with ingestion, due to the plant’s drasticaxative properties, and with contact with leaves, due to risks ofyncope (for all mammals including domestic animals) (Marzoukt al., 2009). Use is contraindicated during pregnancy as the plant isbortifacient (Pottier-Alapetite, 1981; Delazar et al., 2006). Many ofoday’s traditional medicinal uses of the plant are found through-ut history (see the Eber’s papyrus in ca. 1550 BCE Egypt) (Riddle,999) over a large geographical zone from Mauritania to India, evenxtending outside of the plant endemic zone, to Europe (Adams etl., 2009).

The present investigation was undertaken to establish the acuteoxicity study and to evaluate the analgesic and anti-inflammatoryctivities of the reconstituted lyophilized extracts from vegeta-ive organs (roots, stems and leaves) and two different maturationtates of reproductive organs (fruits and seeds).

. Materials and methods

.1. Sampling and identification

Citrullus colocynthis Schrad. plants were collected in August nearedenine, Tunisia in the municipality of Sidi Makhlouf (33◦33N,

0◦27W). The identification was performed according to the floraf Tunisia (Pottier-Alapetite, 1981) and a voucher specimen (C.C-1.01) deposited in the biological laboratory of the Faculty ofharmacy of Monastir.

.2. Extraction protocol

The extraction was performed on 100 g of each fresh organ:oots, stems, leaves and two different stages of maturation of fruitsnd seeds (immature and ripe). Yields of prepared extracts areravimetrically determined. For preparing the aqueous extract,00 g of each fresh organ were ground with a mixer and addedo 500 ml of distilled water. The mixture was allowed to refluxor 30 min, after which the solution was allowed to cool (4 h at◦C). The mixture was then filtered using filter paper (Whatmano.1) under the vacuum of a water pump. The filtrate obtained was

yophilized, yielding the lyophilized aqueous extract.

.3. Animals

Male adult Wistar rats weighing 160–180 g and Swiss albinosice (weighing 18–25 g) of both sex were obtained from Pas-

eur institute (Tunis, Tunisia). They were housed in polypropyleneages and were left for 2 days for acclimatization to animal roomaintained under controlled condition (a 12 h light–dark cycle at

2 ± 2 ◦C) on standard pellet diet and water ad libitum. Before theay of assay, only the Wistar rats were fasted overnight with theree access to water. Housing conditions and in vivo experimentspproved according to the guidelines established by the Europeannion on Animal Care (CFE Council (86/609)). The rats were used

or the anti-inflammatory evaluation of the aqueous extracts whilehe mice were used for the analgesic investigation and for the acuteoxicity testing.

Animals were divided into drug-treated ‘test’ and saline-treatedcontrol’ groups of six or eight animals per group.

.4. Acute toxicity

For acute toxicity, mice were divided into groups of eight ani-als each. One group served as a control and received 0.9% NaCl

lone (10 ml/kg) given intraperitoneally (i.p.), while the remaining

armacology 128 (2010) 15–19

groups were treated with increasing doses of the aqueous extract:50,100, 250, 500, 750, 1000, 1500, 2000, 3000 and 4000 mg/kg (i.p.),respectively.

The mortality rate within a 48 h period was determined and theLD50 was estimated according to the method described by Millerand Tainter (1944).

According to the results of acute toxicity test, doses were chosenfor pharmacological evaluations. After the last observation the micewere killed and the liver, lungs, heart, spleen and kidneys werewithdrawn, weighed and stoked for next evaluations.

2.5. Analgesic activity

Analgesic activity was performed according to the method ofKoster et al. (1959) and assessed by the acetic acid abdominalconstriction test (writhing test)—a chemical visceral pain model.Swiss albinos mice were selected 1 day prior to each test andwere divided into groups of six mice each. One group served asthe control and was pretreated under cutaneously with 10 ml/kgof saline. Another group was pretreated with the reference drug,acetyl salicylate of lysine (ASL), 200 mg/kg, by the same route.The remaining groups were injected intraperitoneally (i.p.) with10 ml/kg of 1% acetic acid solution 30 min after the administrationof different extracts at the doses of 2, 4 and 8 mg/kg except imma-ture fruits to which we decrease doses to 0.1 mg/kg. After acetic acidadministration, the number of writhes was counted during 30 min.Antinociceptive activity was expressed as inhibition percent ofthe usual number of writhes observed in control animals. Thepercentages of inhibition were calculated according to the follow-ing formula: % inhibition = ((number of writhes)control − (number ofwrithes)treated group) × 100/(number of writhes)control.

2.6. Anti-inflammatory activity

The anti-inflammatory activity was assessed on the basis ofinhibition of paw edema induced by the injection of carrageenan(an edematogenic agent) into the subplantar region of the righthind paw of the rat (Winter et al., 1962). Male Wistar rats weredivided into different groups of 8 animals. The control groupreceived 2.5 ml/kg of saline, the standard group received the ref-erence drug (acetyl salicylate of lysine (ASL), 300 mg/kg) and thetest groups received different organ extracts of Citrullus colocynthisat a dose of 1 and 4 mg/kg. Thirteen minutes after intraperitonealadministration of different substances, 0.05 ml of 1% of carrageenansuspension was injected to all animals in the right hind paw.

The paw volume, up to tibiotarsal articulation, was measuredusing a plethysmometer. The measures were determined at 0 h (V0:before edematogenic agent injection) and 1, 2, 3, 4, 5, 6 and 24 hintervals later (VT). The difference between VT (1, 2, 3, 4, 5, 6 and24 h) and V0 was taken as the edema value. The percentages of inhi-bition were calculated according to the following formula: % inhibi-tion = ((VT − V0)control − (VT − V0)treated group) × 100/(VT − V0)control.

2.7. Statistical analysis

Data obtained from animal experiments were expressed asmean ± S.E.M. and as percentage. Results were statistically evalu-ated by ANOVA and using Student’s t-test. p ≤ 0.05 were consideredsignificant.

3. Results

3.1. Toxicity studies

Swiss albinos mice were observed during 48 h and morbid-ity and/or mortality were recorded, if happens, for each group at

B. Marzouk et al. / Journal of Ethnopharmacology 128 (2010) 15–19 17

Table 1Extraction yields (w/w, %) and LD50 of Citrullus colocynthis Schrad. organ aqueous extracts.

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Extraction yields 2.94 2.21 2.7LD50 (mg/kg) 744.1 884.2 553.7

he end of observation period. Due to death index, the LD50 ofll extracts were determined (Table 1). This value is in relationith organs but also with the maturation states. The LD50 were

anged from 95.8 to 3903.2 mg/kg. Stems were evaluated as theess toxic part of Citrullus colocynthis (LD50 = 3903.2 mg/kg). For theruit extracts, immature fruits had the lowest LD50. Immature seedsere as toxic as mature ones. Leaves seem to be the most toxic

rgan with a LD50 equal to 95.8 mg/kg; all the animals treated withhis extract presented feebleness, hypothermia and insufficiencyespiratory. Looking to the higher toxicity of this vegetative organleaves), it was discarded from the analgesic and anti-inflammatorynvestigations.

.2. Analgesic activity

The inhibition percentages of writhing for all extracts are shownn Table 2. The reference drug inhibited 61.88% of the number of

rithing elicited by acetic acid. The analgesic effect was testedor concentrations ranging from 2 to 8 mg/kg, except immatureruits; to which we decrease the concentration to 0.1 mg/kg. Thedministration of all tested extracts induced a dose-dependentntinociceptive activity. The immature fruits and seeds possess the

ighest analgesic properties; the most active of them were imma-ure fruits as well as at 0.1 mg/kg (93.52%). Stem extracts were asctive as those of roots. The lowest activity was observed for riperuits.

able 2nalgesic activity of different Citrullus colocynthis Schrad. aqueous extracts and ref-rence drug (ASL).

Extract Concentration(mg/kg)

Number of writhes Inhibition ofwrithing (%)

Control – 74.33 ± 4.64 –

Immature seeds 2 2.67 ± 0.51*** 96.414 2.00 ± 0.63*** 97.318 0.50 ± 0.54*** 99.33

Ripe seeds 2 4.50 ± 0.54*** 93.954 2.67 ± 0.51*** 96.418 1.00 ± 0.63*** 98.65

Immature fruits 0.1 3.33 ± 0.51*** 93.520.5 2.67 ± 0.81*** 96.412 1.67 ± 0.51*** 97.764 1.33 ± 0.51*** 98.218 0.33 ± 0.51*** 99.55

Ripe fruits 2 66.50 ± 2.25*** 10.594 48.83 ± 1.94*** 30.428 33.50 ± 1.37*** 54.93

Stems 2 55.83 ± 1.47*** 55.534 21.17 ± 1.16*** 71.528 8.83 ± 0.75*** 88.12

Roots 2 7.33 ± 0.51*** 90.134 5.17 ± 0.75*** 93.058 3.67 ± 0.51*** 95.07

Reference drug (ASL) 200 28.33 ± 2.06*** 61.88

alues are expressed as mean ± S.E.M. (N = 6); ASL: acetyl salicylate of lysine.*** p ≤ 0.001 significant from control.

ruits Ripe fruits Leaves Stems Roots

3.04 12.89 11.05 4.56487.6 95.8 3903.2 833.3

3.3. Anti-inflammatory effect

In carrageenan-induced rat paw edema, all extracts produceda reduction of the edema throughout the entire period of obser-vation. The intraperitoneal administration of the aqueous extractsof reproductive organs (fruits and seeds) reduced significantlythe paw edema induced by the noxious agents. This inhibition isless remarkable after the two vegetative organ (roots and stems)aqueous extract injections. For this preliminary assessment ofthe anti-inflammatory activity, the obtained results demonstratethat the reduction of the paw edema vary in a dose-dependentfashion with a maximum attend at 4 mg/kg. On the contrary,at 1 mg/kg, only a moderate (vegetative organs) and appreciably(reproductive organs) activities were noted. With all extracts, 3 hafter carrageenan injection, the anti-inflammatory activity insti-gate unambiguous increasingly to attend the maximum at 6 and24 h, after a little decrease at 5 h. In terms of plant organs, imma-ture organs (fruits and seeds) showed better activity than ripe ones.The inhibition percentages were ranged, respectively, from 90.00%to 98.84% and from 88.33% to 98.06% for immature organs, andranged from 78.33% to 97.69% and from 65.00% to 97.90%, respec-tively, for ripe fruits and seeds. The highest activity was found forimmature fruits. Stem aqueous extracts were either as stronglyanti-inflammatory as root aqueous extract (1–6 h) or a close secondafter 24 h from the carrageenan injection. Standard drug decreasedpaw edema by a maximum of 74.40% after 4 h (Table 3).

4. Discussion

This is the first study evaluating the in vivo acute toxicity, theantinociceptive and the anti-inflammatory activities of extractsfrom different Citrullus colocynthis organs.

Based on the LD50 calculated, the acute administration dosesof all organ aqueous extracts are estimated (1 and 4 mg/kg). Thesedoses which are very slighter than the LD50 were demonstrated effi-cient. The leaf toxicity and its symptoms are confirmed by peoplesfrom Sidi Makhlouf municipality since they suffer from feeblenessand insufficiency respiratory every time they were considerablyprojected to this organ. This expressed toxicity was absolutely notconformed to the traditional use of this plant organ (Batanouny,1999). This contradiction may be attributed to the used doses andpreparations as folkloric medicine. Bruneton (1999) showed thatthe leaf toxicity appear on sheep at a dose of 250 mg/kg (>obtainedLD50 = 95.8 mg/kg). So plant toxicity is in relation with many condi-tions like the geographical distribution and the collect season whichconsiderably influenced the plant composition, and then its toxicityand its pharmacological activity. In addition, it is known that themetabolism and the pharmacological activities are very differentaccording to the animal models (Bertrand, 1976; Rico, 1978).

In acetic acid-induced writhing in mice, all tested samplesextracts reduced significantly the number of writhing which areassociated with the release of endogenous substances includingserotonin, histamine, prostaglandin and bradykinin (Collier et al.,1968). The results obtained in this test thus suggest that whileCitrullus colocynthis Schrad. organs possess peripheral analgesic

properties, this particular activity is probably linked to their anti-inflammatory effects.

Carrageenan has been widely used as a noxious agent able toinduce experimental inflammation for the screening of compoundspossessing anti-inflammatory activity. This phlogistic agent, when

18B.M

arzouket

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128 (2010) 15–19

Table 3Effects of different Citrullus colocynthis Schrad. organ aqueous extracts and reference drug on carrageenan-induced paw edema.

Extract Dose (mg/kg) Mean swelling thickness (10−2) ± S.E.M. (% inhibition)

1 h 2 h 3 h 4 h 5 h 6 h 24 h

Control 1 – 15.00 ± 1.76 24.00 ± 2.66 49.00 ± 4.77 57.50 ± 5.56 59.50 ± 5.25 64.50 ± 8.20 32.50 ± 3.51

Immature seeds 1 14.50 ± 2.89ns (3.33) 19.25 ± 2.21** (19.79) 33.25 ± 2.21*** (32.14) 32.50 ± 2.38*** (43.48) 39.00 ± 3.91*** (34.45) 29.25 ± 3.86*** (54.65) 14.25 ± 0.96*** (56.15)4 1.75 ± 0.96*** (88.33) 2.00 ± 0.82*** (91.66) 3.50 ± 0.58*** (92.86) 1.50 ± 1.29*** (97.40) 2.50 ± 1.29*** (95.80) 1.25 ± 0.50*** (98.06) 0.75 ± 0.95*** (97.69)

Ripe seeds 1 15.00 ± 0.82ns (0.00) 23.75 ± 1.25ns (1.04) 42.00 ± 6.16* (14.28) 37.50 ± 3.87*** (34.78) 42.25 ± 2.22*** (28.99) 29.50 ± 3.41*** (54.26) 14.50 ± 1.29*** (55.38)4 5.25 ± 1.26*** (65.00) 2.25 ± 0.50*** (90.62) 1.75 ± 0.96*** (96.43) 1.25 ± 0.50*** (97.83) 1.50 ± 0.58*** (97.48) 1.50 ± 0.58*** (97.67) 1.25 ± 0.50*** (96.15)

Immature fruits 1 13.25 ± 2.22ns (11.66) 16.75 ± 3.30** (30.21) 29.00 ± 2.16*** (40.82) 31.75 ± 3.86*** (44.78) 34.50 ± 1.29*** (43.70) 21.75 ± 3.95*** (66.02) 16.75 ± 2.22*** (48.46)4 1.50 ± 0.58*** (90.00) 1.75 ± 0.96*** (92.71) 2.75 ± 0.50*** (94.39) 1.75 ± 0.95*** (97.65) 2.75 ± 0.96*** (95.38) 0.75 ± 0.96*** (98.84) 0.50 ± 0.58*** (98.46)

Ripe fruits 1 14.00 ± 1.41ns (6.66) 21.75 ± 1.50* (9.37) 42.75 ± 2.63** (12.75) 43.75 ± 3.09*** (23.91) 50.25 ± 1.70*** (15.55) 43.00 ± 3.83*** (33.33) 17.75 ± 2.05* (45.38)4 3.25 ± 0.96*** (78.33) 2.50 ± 0.57*** (89.58) 3.5 ± 0.57*** (92.86) 4.25 ± 2.63*** (92.61) 4.75 ± 0.50*** (92.02) 1.75 ± 0.50*** (97.29) 0.75 ± 0.50*** (97.69)

Control 2 – 15.00 ± 1.76 34.50 ± 4.33 57.00 ± 6.22 61.50 ± 6.42 67.00 ± 7.37 71.00 ± 6.39 40.50 ± 5.84

Stems 1 14.00 ± 1.41ns (6.66) 31.75 ± 1.25* (7.97) 44.25 ± 2.75*** (22.37) 46.75 ± 3.09*** (23.98) 53.75 ± 2.99*** (19.77) 47.00 ± 1.82*** (33.80) 26.75 ± 1.5*** (33.95)4 13.75 ± 2.21ns (8.33) 30.50 ± 1.91* (11.59) 33.25 ± 2.98*** (41.66) 27.00 ± 3.37*** (56.10) 36.00 ± 2.16*** (46.27) 32.00 ± 2.16*** (54.93) 13.00 ± 1.15*** (67.90)

Roots 1 15.00 ± 2.45ns (0) 34.25 ± 1.70ns (0.72) 55.00 ± 3.74ns (3.51) 51.50 ± 1.29*** (16.26) 59.50 ± 1.29*** (11.19) 49.50 ± 1.91*** (25.00) 28.00 ± 0.82*** (30.86)4 13.75 ± 2.22ns (8.33) 31.50 ± 2.38ns (8.69) 46.50 ± 2.08*** (18.42) 47.00 ± 3.16*** (23.58) 53.00 ± 1.82*** (20.89) 53.25 ± 3.20*** (30.28) 18.25 ± 1.71*** (54.94)

ASL 300 7.25 ± 0.96*** (51.66) 14.25 ± 1.71*** (58.69) 17.50 ± 2.38*** (69.30) 15.75 ± 2.22*** (74.40) 19.50 ± 2.08*** (70.89) 19.50 ± 1.29*** (72.53) 23.75 ± 1.71*** (41.36)

Values are expressed as mean ± S.E.M. (N = 8); ns: not significant from the control; ASL: acetyl salicylate of lysine.* p ≤ 0.05 significant from the control.

** p ≤ 0.01 significant from the control.*** p ≤ 0.001 significant from the control.

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njected locally into the rat paw, produced a severe inflammatoryeaction, which was discernible within 30 min (John and Nodine,999). The development of edema induced by carrageenan is aiphasic event: the early phase (90–180 min) of the inflammation

s due to the release of histamine, serotonin and similar substances.he later phase (270–360 min) is associated with the activation ofinin-like substances and the release of prostaglandins, proteasesnd lysosome (Olajide et al., 1999). All organ extracts inhibitedind paw edema and showed a dose-depending anti-inflammatoryctivity but the results were different for each organ depend-ng on the early/later phases. Extracts from reproductive organsseeds and fruits: immature and ripe ones) inhibited the bothhases of the carrageenan-induced edema by reducing the releasef histamine and serotonin and also the kinin-like substances androstaglandins; while the extracts from vegetative organs (rootsnd stems) inhibited particularly the later phases (4–24 h) byestraining the kinin-like substances and prostaglandins produc-ions. This pharmacological property may be attributed to the plantrgan composition and to a possible molecular mechanism by effec-ively decreasing the production of the pro-inflammatory cytokinesf IL-6 and IL-1� and the expression of COX-2 and simultane-usly elevating the level of anti-inflammatory cytokine IL-4 in thearrageenan-injected rat paw tissues (Moulin and Coquerel, 2002).

At any rate, these results indicate that the analgesic and anti-nflammatory activities could not be imputed to one family ofhytochemicals only (or its absence). Like for the antibacterialnd the anti-candidal activities, analgesic and anti-inflammatorynes may be attributed, possibly in combination, to various phy-ochemicals detected (alkaloids, iridoids, flavonoids, steroids, etc.)Marzouk et al., 2009). These compounds might be present in vari-us concentrations according to the maturation stage of seeds andruits (Marzouk et al., 2009). Alkaloids are commonly found to havenalgesic and anti-inflammatory properties (Moulin and Coquerel,002); therefore their absence in roots could account for the lack ofctivity of this organ. Iridoids, witch have anti-inflammatory effectsMesia-Vela et al., 2004), are detected in the aqueous root extractut this extract showed no overall activity. However, alkaloids and

ridoids cannot be solely responsible for the activity. Flavonoidsdetected in all the seed extracts) are known to have analgesic andnti-inflammatory properties (Borgi et al., 2008); the same coulde said about steroids witch contribute to the better performances an anti-inflammatory agent (Bames and Adcock, 2009).

With these analgesic and anti-inflammatory properties, Citrul-us colocynthis Schrad. can be considered an effective agent toreat inflammation diseases. This plant, namely its seed and fruitxtracts, demonstrated a high activity at very low aqueous extractoses (1 and 4 mg/kg). The study corroborated the analgesic effectsf this specie, justified and supported scientifically its ethnophar-acological use as an anti-inflammatory agent to treat pain and

heumatoid arthritis. Therefore it could account for some of theariations observed in the Ethnopharmaceutical preparation meth-

ds. From now the use of this plant is validated by the resultsbtained in this work. Additional studies are ongoing to confirm thisitrullus colocynthis Schrad. properties with some ecological vari-bilities. Further attempts to isolate and define the active analgesicnd anti-inflammatory fractions and its components.

armacology 128 (2010) 15–19 19

Acknowledgements

Thanks to Dr. Zohra MARZOUK for the specimen identifica-tion and to Dr. Wahida BORGI and Dr. Rachel DECOR for theirhelp.

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