COLIMIL ON UPPER GASTROINTESTINAL TRANSIT 999

Copyright © 2007 John Wiley & Sons, Ltd. Phytother. Res. 21, 999–1001 (2007)DOI: 10.1002/ptr

Copyright © 2007 John Wiley & Sons, Ltd.

PHYTOTHERAPY RESEARCHPhytother. Res. 21, 999–1001 (2007)Published online 20 June 2007 in Wiley InterScience(www.interscience.wiley.com) DOI: 10.1002/ptr.2192

SHORT COMMUNICATIONEffects of the Herbal Formulation ColiMil® onUpper Gastrointestinal Transit in Mice In Vivo

Raffaele Capasso1*, Francesco Savino2 and Francesco Capasso1

1Department of Experimental Pharmacology, University of Naples Federico II, 80131 Naples Italy2Department of Pediatrics, University of Turin, Ospedale Infantile Regina Margherita, Torino 10126, Italy

Clinical evidence suggests that the herbal formulation ColiMil® (which contains Matricaria recutita flowersextract, Foeniculum vulgare fruit extract and Melissa officinalis aerial parts extract) is effective in the treat-ment of breastfed colic in infants. Therefore the effect of this phytotherapeutic formulation and its herbalconstituents on upper gastrointestinal transit was investigated in mice in vivo. Oral administration of theherbal formulation (0.4–0.8 mL/mice) dose-dependently delayed upper gastrointestinal transit. Among theherbal components, Matricaria recutita extract (0.89 and 1.78 mg/mouse) and Melissa officinalis extract(6.46 and 12.92 mg/mouse), but not Foeniculum vulgare (8.21 and 16.42 mg/mouse), reduced motility signifi-cantly. These results suggest that ColiMil® reduces upper gastrointestinal motility in mice, with a majorcontribution by Matricaria recutita and Melissa officinalis. These experimental data may be important tobetter understand the observation that the herbal formulation ColiMil® improves colic in breastfed infants.Copyright © 2007 John Wiley & Sons, Ltd.

Keywords: Matricaria recutita; Melissa officinalis; Foeniculum vulgare; intestinal motility.

INTRODUCTION

Although infantile colic is one of the most commonproblems of infants within the first 3 months of life, thepathophysiology of this clinical entity characterized byexcessive crying is not completely understood and itsdiagnosis is still based on Wessel’s diagnostic criteria(Wessel et al., 1954). An hypothesis includes abnormalintestinal motility that can be treated using conven-tional spasmolytic therapies (Weissbluth et al., 1984;Savino et al., 2002). More recently Savino et al. (2005)have shown that colic in breastfed infants improveswithin a week of treatment with a herbal formulationcontaining three plant extracts: Matricaria recutita,Foeniculum vulgare and Melissa officinalis. Both chamo-mile and fennel are traditionally used for the sympto-matic treatment of gastrointestinal disturbances such asimpaired digestion, flatulence, epigastric bloating andas an adjunctive treatment of the painful component offunctional dyspepsia (Forster et al., 1980; Westpal andHorning, 1996). An extensive literature exists also onthe spasmolytic effect of lemon-balm (Forster et al.,1980; Soulimani et al., 1991; 1993). The experimentalpharmacology of the above mentioned herbal formula-tion is largely unexplored and, in particular, it is not

known if it affects intestinal motility. To evaluate apossible synergistic effect among the different plantextracts the effects of the herbal formulation ColiMil®

and of single plant extracts on upper gastrointestinaltransit in mice in vivo were compared, both in physio-logical and pathophysiological states. Some of theseresults have been presented at the European Academyof Paediatrics, Barcelona, 2006 (Capasso et al., 2006).

METHODS

Animals. Male ICR mice (Harlan Italy, S. PietroalNatisone, UD) (26–30 g) were used after 1 week ofacclimation (temperature 23 ± 2 °C; humidity 60%).Food was withheld 6 h before transit measurementand 18 h before the induction of chronic intestinalinflammation. All animal experiments complied withthe Italian D.L. no. 116 of 27 January 1992 and associ-ated guidelines in the European Communities CouncilDirective of 24 November 1986 (86/609/ECC).

Upper gastrointestinal transit. Gastrointestinal transitwas measured in control and in croton oil-treated mice.At this time, 0.1 mL/10 g mouse of a black marker (10%charcoal suspension in 5% gum arabic) was admini-stered orally to assess upper gastrointestinal transit aspreviously described (Puig and Pol, 1998). After 20 minthe mice were killed by asphyxiation with CO2 and thegastrointestinal tract removed. The distance travelledby the marker was measured and expressed as a per-centage of the total length of the small intestine frompylorus to caecum.

Received 26 March 2007Accepted 30 March 2007

* Correspondence to: Dr Raffaele Capasso, Department of ExperimentalPharmacology, University of Naples Federico II, Via D. Montesano 49,80131 Naples, Italy.E-mail: rafcapas@unina.itContract/grant sponsor: Milte Italia SpA.

Copyright © 2007 John Wiley & Sons, Ltd. Phytother. Res. 21, 999–1001 (2007)DOI: 10.1002/ptr

1000 R. CAPASSO ET AL.

Intestinal inflammation. Inflammation was induced aspreviously described (Puig and Pol, 1998). Mice receivedorally two doses of croton oil (20 μL/mouse) on 2 con-secutive days. Motility was measured 4 days after thefirst administration of croton oil. This time was selectedon the basis of previous work (Puig and Pol, 1998),which reported that the maximal inflammatory responseoccurred 4 days after the first treatment.

Drug administration. ColiMil® (0.4 and 0.8 mL/mouse),Matricaria recutita flower extract (0.89 and 1.78 mg/mouse), Foeniculum vulgare fruit extract (8.21 and16.42 mg/mouse), Melissa officinalis aerial part extract(6.46 and 12.92 mg/mouse) or loperamide (10 mg/kg)were given per os 60 min before charcoal admini-stration. In another set of experiments herbal ColiMil®

(0.4–0.8 mL/mouse) or loperamide (10 mg/kg) weregiven per os (60 min before charcoal administration) incroton oil-treated mice.

Drugs. ColiMil® is an herbal formulation containingMatricaria recutita flower methanol extract, standard-ized to apigenin (0.3%), Foeniculum vulgare fruitwater extract, standardized to essential oil (0.05–0.1%)and Melissa officinalis aerial part water extract, stand-ardized to rosmarinic acid (1.5%). Each millilitre ofColiMil® (a gift from Milte-Milan, Italy) contained:2.225 mg of Matricaria recutita extract, 20.537 mg ofFoeniculum vulgare extract and 16.15 mg Melissaofficinalis extract. Loperamide and croton oil were pur-chased from Sigma (Milan, Italy). All the drugs (withthe exception of croton oil) were dissolved in water.

Statistical analysis. Data are mean ± SEM. To deter-mine statistical significance, Student’s t-test for unpaireddata or one-way analysis of variance followed by Tukey-Kramer multiple comparisons test was used. A value ofp < 0.05 was considered significant.

RESULTS AND DISCUSSION

Charcoal meal administered intragastrically to controlanimals covered 49 ± 3% of the total length of thesmall intestine in 20 min. The herbal formulation tested(0.4 and 0.8 mL/mouse, orally) dose-dependently andsignificantly inhibited gastrointestinal transit (Fig. 1).Among the components, Matricaria recutita flowerextract (0.89 and 1.78 mg/mouse) and Melissa officinalis(6.46 and 12.92 mg/mouse) reduced significantly motil-ity. Foeniculum vulgare fruit extract (8.21 and 16.42 mg/mouse), did not show consistent responses (Fig. 1).

Administration of croton oil evoked a significantincrease in gastrointestinal transit which was reversedin mice treated with the herbal formulation (Fig. 2).Pretreatment with the herbal formulation (0.4 and0.8 mL/mouse per os) counteracted the increase ingastrointestinal transit induced by croton oil in adose-dependent fashion (Fig. 2).

Loperamide (10 mg/kg, orally), used as a referencedrug, inhibited the gastrointestinal transit both in con-trol mice (% transit: control: 52 ± 3; loperamide: 35 ± 4;n = 6, p < 0.01) and in mice treated with croton oil(% transit: croton oil: 65 ± 4; croton oil + loperamide:23 ± 4; n = 6, p < 0.001).

Figure 1. Inhibitory effect of Foeniculum vulgare fruit extract(FV, 8.21 and 16.42 mg/mouse); Matricaria recutita flower ex-tract (MR, 0.89–1.78 mg/mouse, per os); Melissa officinalis aerialpart extract (MO, 6.46–12.92 mg/mouse, per os); herbal formu-lation (ColiMil®: Matricaria recutita flower extract, Foeniculumvulgare fruit extract and Melissa officinalis aerial part extract;0.4–0.8 mL/mouse, per os) on gastrointestinal transit. Resultsare mean ± SEM of 14–16 animals for each experimental group.* p < 0.05, ** p < 0.01, *** p < 0.001 vs control.

Figure 2. Croton oil-treated mice: inhibitory effect of herbal for-mulation (ColiMil®: Matricaria recutita flower extract, Foeniculumvulgare fruit extract and Melissa officinalis aerial part extract;0.4–0.8 ml/mouse, per os) on gastrointestinal transit. Resultsare mean ± SEM of 10–11 animals for each experimental group.# p < 0.05 vs control; * p < 0.05 vs inflammation.

COLIMIL ON UPPER GASTROINTESTINAL TRANSIT 1001

Copyright © 2007 John Wiley & Sons, Ltd. Phytother. Res. 21, 999–1001 (2007)DOI: 10.1002/ptr

Based on the results, it can be seen that the animalgroup treated exclusively with Melissa officinalis orMatricaria recutita, but not with Foeniculum vulgare,showed a significant reduction of the motility comparedwith the control group. The decrease was greater in thegroup simultaneously treated with the herbal formula-tion. When the percent inhibition was calculated withreference to the control group, the results showed amean reduction of 28% in the group treated withchamomile, 36% in the group treated with lemon balm,9% in the group treated with fennel and 45% in thegroup treated with ColiMil®. To our knowledge, thisis the first systematic report directly demonstrating aneffect of a combination of herbal formulation on intesti-nal motility both in physiological and pathophysiological(inflamed) conditions. Possible interactions (synergisms)among the single constituents and with specific enzymesand mediators involved in gut motility may be taken

into consideration to explain the superiority of the phar-macological profile of the herbal formulation over thesingle extracts. Nevertheless, the precise mechanisminvolved in this synergy effect and the relative contri-bution of the different components of ColiMil® awaitfurther study. In conclusion the data demonstrate thatthe herbal formulation containing Matricaria recutita,Foeniculum vulgare and Melissa officinalis reduced gutmotility in mice. The inhibitory effect here observedmight explain, at least in part, the clinical efficacy ofthis herbal formulation in the treatment of breastfedcolicky infants (Weizman et al., 1993; Alexandrovichet al., 2003; Savino et al., 2005).

Acknowledgements

This work was supported by Milte Italia SpA.

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