European Journal of Pharmacology 569 (2007) 197 203 www.elsevier.

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Antidepressant-like effect of hyperfoliatin, a polyisoprenylated phloroglucinol derivative from Hypericum perfoliatum (Clusiaceae) is associated with an inhibition of neuronal monoamines uptake
Jean-Claude do Rego a,, Nama Benkiki b , Elizabeth Chosson c , Zahia Kabouche b , Elisabeth Seguin c , Jean Costentin a
a CNRS FRE 2735, Laboratory of Experimental Neuropsychopharmacology, European Institute for Peptide Research (IFRMP 23), Faculty of Medicine and Pharmacy, Institute of Biomedical Research, University of Rouen, 22 Boulevard Gambetta, 76183 Rouen Cedex, France b Laboratoire d'Obtention de Substances Thrapeutiques (LOST), Faculty of Sciences, University of Mentouri - Constentine, Campus Chaabet Ersas, 25000 Constantine, Algeria c CNRS UMR 6014, Laboratory of Pharmaceutical Chemistry, Faculty of Medicine and Pharmacy, University of Rouen, 22 Boulevard Gambetta, 76183 Rouen Cedex, France

Received 29 December 2006; received in revised form 2 May 2007; accepted 3 May 2007 Available online 13 May 2007

Abstract This study investigated, in mice, the antidepressant like effect of hyperfoliatin, a prenylated phloroglucinol derivative isolated from the aerial parts of Hypericum perfoliatum, as well as its action on monoaminergic systems. In the forced-swimming test, hyperfoliatin dose-dependently reduced immobility time. Immobility was interpreted as an expression of behavioural despair, which could be a component of depression syndrome. The effect of hyperfoliatin did not result from the stimulation of animal motor activity. Hyperfoliatin inhibited, in a concentrationdependent manner, the [3H]-dopamine, [3H]-serotonin and [3H]-noradrenaline synaptosomal uptakes, but did not prevent the binding of specific ligands to the monoamine transporters. These data suggest that the antidepressant-like effect of hyperfoliatin on the forced-swimming test is probably associated to monoamine uptake inhibition, due to a mechanism of action different from that of known antidepressants. 2007 Elsevier B.V. All rights reserved.
Keywords: Antidepressant drug; Monoaminergic system; Forced-swimming test; Hypericum perfoliatum; Hyperfoliatin

1. Introduction The genus Hypericum L. (Clusiaceae) includes numerous species which have been used as medicinal plants for centuries in the treatment of trauma, burns, rheumatism, neuralgia, gastroenteritis, ulcers, hysteria, bedwetting and depression (Miller, 1998). In the past two decades several studies have demonstrated that extracts of Hypericum species contain antidepressant properties, as the tricyclic antidepressant, in humans (Ernst, 1995; Linde et al., 1996; Volz, 1997; Stevinson and Ernst, 1999) and exert an antidepressant-like action in laboratory animals (Bhattacharya et al., 1998; Butterweck et al., 1998; Mller et al., 1998; Panocka et al., 2000; Daudt et al., 2000; Sanchez-Mateo et al., 2002;
Corresponding author. Tel.: +33 2 35148602; fax: +33 2 35148603. E-mail address: jean-claude.dorego@univ-rouen.fr (J.-C. do Rego). 0014-2999/$ - see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.ejphar.2007.05.008

Mller, 2003; Viana et al., 2005). However, neither the mechanisms of action nor the identity of the active constituents has yet been completely understood. Naphtodianthrone hypericin has been considered as the most active constituent of European St John's wort, Hypericum perforatum L. (Butterweck et al., 1997). In fact, some reports still provide evidence for the role of hypericin in antidepressant activity of H. perforatum (Butterweck et al., 1998) and the majority of the phytomedicines available are standardized on this naphthodianthrone. Nevertheless, currently most researchers consider that the antidepressant effects are due to a variety of constituents rather than a single one (Chatterjee et al., 1998a; Butterweck et al., 2000). Xanthones (Rocha et al., 1994), flavonoids (Butterweck et al., 2000) and in particular phloroglucinol derivatives (Chatterjee et al., 1998b; Mller et al., 2001; Viana et al., 2005) are also considered as pharmacologically relevant compounds. Among these extensively studied

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from both chemical and pharmacological points of view, their biological properties appear to be correlated with their prenylation, thus hyperforin (Bystrov et al., 1975; Beerhues, 2006) and adhyperforin (Maisenbacher and Kovar, 1992) have emerged as the constituents responsible for the antidepressant activity of H. perforatum (Chatterjee et al., 1998b; Barnes et al., 2001; Jensen et al., 2001; Mller et al., 2001; Roz et al., 2002). Recently, we reported the structure determination of hyperfoliatin, a novel prenylated phloroglucinol derivative, isolated from the aerial parts of Hypericum perfoliatum L. locally used in folk medicine for wound healing and in the treatment of various bacterial diseases (Benkiki et al., 2003). Hyperfoliatin was also extracted from aerial parts of Hypericum scabrum, an Usbekistan medicinal plant (Tanaka et al., 2004). In this study we further investigated about the hyperfoliatin antidepressant-like effect and its mechanism of action. For this purpose, we tested the hyperfoliatin potential antidepressant effect on an experimental model of depression, the forcedswimming test, then we attempted to assess its effects on the uptake complex of monoaminergic systems. 2. Materials and methods 2.1. Behavioural experiments 2.1.1. Animals Male Swiss albinos CD1 mice (IFFA-CREDO/Charles River, Saint-Germain sur L'Arbresle, France), weighing 2530 g, were housed 20 in Makrolon cages (L: 40 cm, W: 25 cm, H: 18 cm), with free access to water and food (U.A.R., Villemoisson sur Orge, France). The animals were kept in a ventilated room, at a temperature of 22 1 C, under a 12-h light/12-h dark cycle (lights on between 7:00 a.m. and 7:00 p.m.). All the experiments were carried out between 9:00 a.m. and 6:00 p.m., in testing rooms adjacent to the animal stabulation rooms. Animal manipulations were performed according to the European Communities Council Directive of 24 November 1986 (86:609:EEC) and conducted by authorized investigators. Each animal was used once, then immediately sacrificed. 2.1.2. Drug and solution Hyperfoliatin was diluted to a concentration of 80 mg/kg in saline with 2% cremophor and 2% DMSO, and then diluted in saline immediately before treatment. 2.1.3. Forced-swimming test The forced-swimming test was essentially similar to that described by Porsolt et al. (1977), but used an apparatus with a larger Plexiglas cylinder (14 cm in diameter, instead of 10 cm) similar to that employed by Semba and Takahashi (1988) and DoRego et al. (2002, 2005), since Sunal et al. (1994) have established that a cylinder with a higher diameter decreases the number of false positive responses. The apparatus consisted of two Plexiglas cylinders (20 cm height, 14 cm internal diameter), placed side-byside in a Makrolon cage (L: 38, W: 24, H: 18 cm) and separated by an opaque screen. The Makrolon cage was filled with water at 22

1 C, to a height of 12 cm instead of 6 cm as suggested by Porsolt et al. (1977), since, according to Petit-Demouliere et al. (2005), the depth of water is an important parameter to be considered as mice should not sense a limit under the level of water. The behaviour of the mice would indeed be altered if their tails touch the bottom of the cylinder. Fifteen minutes before the test the animals were isolated in small individual cages (L: 25, W: 9, H: 8 cm) at an ambient temperature (22 1 C). Thirty or 60 min after i.p. injection with vehicle (saline containing 0.05% cremophor + 0.05% DMSO) or graded doses of hyperfoliatin, four mice were tested simultaneously for a 6-min period. Total duration of immobility was measured during three consecutive periods of 2 min, each using an automated image analysis system (Videotrack MV 45). The method was approved by the Regional Ethical Committee for Animal Experimentation (Normandy; no. N/09-04-04-11). 2.1.4. Measurement of locomotor activity Locomotor activity was measured in a Digiscan actometer (Omnitech Electronics Inc, Colombus, OH, U.S.A.), which monitored the horizontal and the vertical movements of the animals. The animals were placed individually in 20 20 30 cm compartments, placed into a dimly lit and quiet room. The recording apparatus was connected to a Compaq Computer in order to process the data. The responses to hyperfoliatin, injected intraperitoneally immediately before the test, were expressed as the number of crossed beams, during four consecutive 15 min periods. 2.2. In vitro assays 2.2.1. Animals Male Sprague Dawley rats, weighting 180200 g, were purchased from IFFA-CREDO/Charles River Laboratories (Domaine des Oncins, Saint-Germain sur L'Arbresle, France). They were housed by 4 in Makrolon cages (L: 40 cm, W: 25 cm, H: 18 cm), with free access to water and food (U.A.R., France) and kept in a well ventilated room, at a temperature of 21 1 C, under a 12-h light dark cycle (lights on between 7-h and 19-h). The procedures used in this study are in compliance with the European Communities Council Directive of 24 November 1986 (86/609/EEC). 2.2.2. Drugs [3H]-dopamine (48 Ci/mmol) and [3H]-nisoxetine (86 Ci/ mmol) were purchased from Amersham (Les Ulis, France). [3H]noradrenaline (12.5 Ci/mmol), [3H]-serotonin (25.5 Ci/mmol), [3H]-citalopram (84.2 Ci/mmol) and [3H]-mazindol (24.5 Ci/ mmol) were purchased from Perkin Elmer-NEN Life Science Products (Paris, France). Cocaine hydrochloride was obtained from la Cooprative Pharmaceutique Franaise (Melun, France). Desipramine hydrochloride, fluoxetine hydrochloride and GBR 12783 were purchased from Sigma-Aldrich (Saint Quentin Fallavier, France). Details of extraction, isolation and identification of hyperfoliatin have previously been reported (Benkiki et al., 2003). Solutions of hyperfoliatin (10 410 10 M) were prepared in an incubation medium containing 0.05% cremophor and 0.05% DMSO. Millimolar solutions of GBR 12783 were prepared in

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distilled water. Subsequent dilutions and solutions of other agents were performed in the incubation medium. 2.2.3. Synaptosomal uptake of [3H]-dopamine, [3H]-serotonin and [3H]-noradrenaline All procedures necessary to prepare synaptosomal suspensions were performed at 02 C. Animals were sacrificed by decapitation; their striata (for [3H]-dopamine uptake), frontal cortex (for [3H]-serotonin uptake) and hypothalamus (for [3H]noradrenaline uptake) were dissected out and homogenized with 12 up and down strokes of a teflon-glass homogenizer (800 r.p.m.) in 10 volumes (w/v) of ice-cold 0.32 M sucrose solution containing 0.1 mM pargyline. The nuclear material was removed by centrifugation at 1000 g for 10 min, and the supernatant (crude synaptosomal fraction) was used in uptake experiments. Aliquots (100 l) of the crude synaptosomal fraction were preincubated for 5 min at 37 C in a KrebsRinger medium, containing (mM): NaCl 109, KH2PO4 1, CaCl2 1, NaHCO3 27, glucose 5.4, pH 7.4 0.1, in presence or absence of hyperfoliatin. The incubation was continued for 5 min in the same medium, in the presence of [3H]-dopamine, [3H]-serotonin or [3H]noradrenaline (10 nM; 1 ml final volume). The reaction was stopped by adding 3 ml of ice-cold incubation medium and an immediate centrifugation (7000 g, 10 min, 4 C). The pellet was washed with 1 ml of the latter medium and centrifuged under the same conditions. The final pellet was sonicated in 250 l distilled water and aliquots of the homogenate were used for the determination of radioactivity and protein concentrations. The radioactivity was determined by liquid scintillation spectrometry (Betamatic V, KONTRON, Trapes, France) in 4 ml of Optiphase Highsafe II with 3336% counting efficiency. Protein concentrations were determined according to the method of Lowry et al. (1951), using bovine serum albumin as standard. The specific uptake of dopamine, serotonin or noradrenaline was defined as the difference between the total uptake at 37 C and the non-specific accumulation observed at 0 C, respectively in the presence of 100 M cocaine for [3H]-dopamine, 1 M fluoxetine for [3H]-serotonin or 0.3 M desipramine for [3H]noradrenaline. [3H]-serotonin and [3H]-noradrenaline uptake assays were performed in the presence of 30 nM 1-[2(diphenylmethoxy)ethyl]-4-(3-phenyl-2-propenyl)-piperazine, dihydrochloride (GBR 12783) in order to block serotonin and noradrenaline transporters operated by dopamine nerve terminals present respectively in cortical and hypothalamic preparations. The monoamine synaptosomal uptake was analysed using the equation of the sigmoidal doseresponse curve (variable slope). IC50 and n Hill values were calculated using the equation of the curve-fitting programs Microcal Origin (Microcal Software), under-mentioned below: h  i Y Bottom Top Bottom= 1 X =IC50 nHill 2.2.4. Binding assays to monoamine transporters The binding to dopamine (DAT), noradrenaline (NAT) or serotonin (SERT) transporters was assessed, respectively, on striatal, cortex and hypothalamus membranes of rats. The crude

synaptosomal suspensions, prepared as described above, were centrifuged at 17,000 g for 30 min at 4 C. The pellet was resuspended by sonication in: 10 mM Na+ medium (0.30 mM NaH2PO4, 9.70 mM NaHCO3, pH 7.5 0.1) for the binding to DAT or 50 mM Tris HCl (containing 120 mM NaCl and 5 mM KCl, pH 7.4 0.1) for NAT and SERT; and centrifuged (50,000 g, 10 min, 4 C). The final pellet was resuspended by sonication in the same respective medium for the binding to DAT and SERT. For NAT it was resuspended in 50 mM Tris HCl (containing 300 mM NaCl and 5 mM KCl, pH 7.4 0.1). [3H]-Mazindol (2 nM final concentration) and membranes (100 g protein) were incubated at 0 C, for 2 h, to evaluate the binding to DAT. For the binding to NAT 300 g of protein were incubated with [3H]-nisoxetine (1 nM) at 25 C during 2 h; and for SERT, 200 g of protein were incubated with [3H]citalopram (1 nM) during 1 h at 25 C. All incubations were carried out in the presence of different hyperfoliatin

Fig. 1. Doseresponse effect of hyperfoliatin on the forced-swimming test. 30 min (A) or 60 min (B) after the treatment, mice were submitted to the swimming test, and the duration of immobility was measured during a 6-min period (three consecutive periods of 2 min each). Data represent mean S.E.M. of 10 mice per group. A two-way ANOVA failed to reveal significant interactions between treatment and time after treatment: F(3,72) = 0.58, P = 0.63 at 120 min; F(3,72) = 1.05, P = 0.37 at 240 min; F(3,72) = 0.59, P = 0.62 at 360 min. Student NewmanKeuls test: P b 0.05, P b 0.01, P b 0.001 vs. respective control.

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concentrations (1 10 63 10 11 M), in a final volume of 500 l. Incubations were stopped by dilution with 3 ml of icecold incubation medium and immediate filtration through GF/B filters, previously soaked, for at least 1 h, in 0.5% polyethyleneimine. Each tube and filter was rinsed once or twice with 3 ml of ice-cold incubation medium. Filters were counted for radioactivity by liquid scintillation spectrometry (Tri-carb 2100TR, Packard BioScience Co.) in 4 ml of Ultima Gold (Perkin Elmer). The non-specific binding was determined by incubation with cocaine (100 M, for DAT), desipramine (10 M, for NAT) or fluoxetine (10 M, for SERT). Protein concentrations were determined according to the method of Lowry et al. (1951), using bovine serum albumin as standard. 3. Results 3.1. Behavioural experiments As shown in Fig. 1, 30 or 60 min after treatment, hyperfoliatin (5 to 20 mg/kg i.p.) induced a dose-dependent decrease in the
Fig. 3. Effect of hyperfoliatin on the in vitro uptake of [3H]-dopamine, [3H]noradrenaline and [3H]-serotonin. Aliquots of synaptosomal suspensions obtained from rat striatum ([3H]-dopamine), hypothalamus ([3H]-noradrenaline) or frontal cortex ([3H]-serotonin) were incubated in the presence of increasing concentrations of hyperfoliatin as described in Materials and methods. Hyperfoliatin-induced uptake inhibition is expressed as percentage of respective basal values. Data are mean S.E.M. of 4 independent experiments performed in duplicate.

immobility time during each of the three 2-min periods of testing and, consequently, during the total duration of the test. The effect of hyperfoliatin was considered significant for dose 20 mg/kg (P b 0.05) at 30 min (Fig. 1A); and for doses 10 mg/kg (P b 0.05) and 20 mg/kg (P b 0.050.001) at 60 min (Fig. 1B) after i.p. treatment. As shown in Fig. 2, the acute treatment with hyperfoliatin (5 to 20 mg/kg) did not significantly modify either horizontal (Fig. 2A) or vertical (Fig. 2B) locomotor activities in any time period of the test. 3.2. Neurochemical experiments 3.2.1. Effects of hyperfoliatin on the uptake of biogenic amines Hyperfoliatin inhibited [3H]-dopamine, [3H]-serotonin and 3 [ H]-noradrenaline uptake operated by crude synaptosomal suspensions prepared from rat striatum (for dopamine), rat frontal cortex (for serotonin) and hypothalamus (for noradrenaline), in a concentration-dependent and monophasic manner (Fig. 3). The IC50 values with respective Hill coefficients (n Hill), which were closed to unity, are summarized in Table 1.

Table 1 Comparison of the inhibition of monoamine uptake by hyperfoliatin Uptake of [3H]-dopamine Fig. 2. Doseresponse effect of hyperfoliatin on horizontal (A) and vertical (B) locomotor activity. Mice were injected intraperitoneally with solvent or hyperfoliatin (520 mg/kg) and placed in the actimeters immediately after the injection. Horizontal displacements and vertical movements were measured for four consecutive periods of 15 min. Data represent mean S.E.M. of 10 mice per group. IC50 (M) 1.2 0.3 n Hill 1.4 0.2 [3H]-serotonin IC50 (M) 3.5 0.99 n Hill 1.2 0.2 [3H]-noradrenaline IC50 (M) 1.8 0.05 n Hill 0.9 0.2

Data were calculated using the equation of the curve-fitting programs Microcal Origin. IC50 and n Hill values represent the mean S.E.M. of 4 independent experiments, carried out in duplicate.

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Fig. 4. Effect of hyperfoliatin on the binding of [3H]-mazindol, [3H]-nisoxetine and [3H]-citalopram. Aliquots of membrane preparations obtained from rat striatum ([3H]-mazindol), hypothalamus ([3H]-nisoxetine) or frontal cortex ([3H]-citalopram) were incubated in the presence of increasing concentrations of hyperfoliatin (3 10 11 to 10 6 M) as described in Materials and methods. A specific binding was expressed as a percentage of that measured in respective control binding without hyperfoliatin. Data are mean S.E.M. of 4 experiments performed in duplicate. The average control specific binding was: 4130 505 fmol/mg prot. (for [3H]-mazindol), 112 16 mg/kg prot. (for [3H]nisoxetine) and 321 53 fmol/mg prot (for [3H]-citalopram).

3.2.2. Effects of hyperfoliatin on the binding of uptake inhibitors to monoamine transporters The different tested concentrations of hyperfoliatin did not inhibit, in a statistically significant manner, the binding of [3H]mazindol, [3H]-nisoxetine and [3H]-citalopram to their respective monoamine transporters (Fig. 4). 4. Discussion In the present study, we demonstrated that hyperfoliatin, a prenylated phloroglucinol derivative, recently isolated from the aerial parts of H. perfoliatum (Benkiki et al., 2003), developed, after an i.p. administration, antidepressant-like effects when assessed in the forced-swimming test. Indeed, it reduced significantly the immobility time in this test. In most behavioural models of depression animals are exposed to mildly aversive situations, from which there is no possibility to escape and which also induces recognizable behavioural changes. In forced-swimming test, a prolonged exposure to an aversive situation induces immobility, interpreted as an expression of behavioural despair, which could also be related to a depression syndrome (Porsolt et al., 1977). A wide variety of antidepressants, including tricyclics, serotonin-specific reuptake inhibitors and monoamine oxidase inhibitors, and other compounds with potential antidepressant-like activity reduce the duration of immobility in the forced-swimming test (Porsolt et al., 1977, 1978; Da Silva et al., 2000; Zomkowski et al., 2002; Cryan et al., 2002; Rosa et al., 2003). In our study, hyperfoliatin also significantly reduced the time of animal immobility in the forced-swimming test. This effect was dose-dependent and had a relatively slow onset. In fact, the

most significant response was observed 60 min after treatment. The magnitude of the effect induced by hyperfoliatin was comparable to that observed after the treatment with potential antidepressant-like activity in the forced-swimming test (PetitDemouliere et al., 2005). The present behavioural observation is in good agreement with the previous studies, showing that phloroglucinol derivatives such as: hyperforin from H. perforatum (Chatterjee et al., 1998b; Mller et al., 2001) and Hypericum triquetrifolium (Roz et al., 2002), as well as HCl from Hypericum caprifoliatum (Viana et al., 2005, 2006) possesses antidepressant-like activities. Furthermore, this anti-immobility effect was not related to a non-specific behavioural stimulation, since hyperfoliatin increased neither the horizontal nor vertical activities evaluated in a new environment. This is in good agreement with the classical observations that antidepressants reduce the immobility in the forced-swimming test at doses which did not stimulate locomotion (Porsolt et al., 1977) and even tended to decrease locomotor activity (Duterte-Boucher et al., 1988; Bourin, 1990). Some studies have indicated that Hypericum species and their phloroglucinol constituents act via a blockade of monoamines uptake into rat brain synaptic vesicles (Roz et al., 2002), as well as into rat synaptosomes (Chatterjee et al., 1998a,b; Mller et al., 1998; Jensen et al., 2001; Viana et al., 2005), and it has been suggested that this effect might explain its antidepressant properties. Based on these data, we demonstrated that hyperfoliatin potently inhibits [3H]-dopamine, [3H]-serotonin and [3H]noradrenaline in synaptosomal preparations, with approximately similar IC50. Contrary to all other known antidepressant drugs, the inhibition of monoamine uptake operated by hyperfoliatin is not due to binding or blockade of dopamine, serotonin or noradrenaline transporters, at the binding sites of [3H]-mazindol, [3H]-citalopram, [3H]-nisoxetine, respectively. These observations suggest that the mechanism of action of hyperfoliatin is probably not associated with a specific binding on each of the different monoamine transporters, but could depend on mechanisms involved in neurotransmitter transport in general. This assumption had been previously reported for extracts of various Hypericum species, such as H. perforatum (Mller et al., 1997, 1998; Chatterjee et al., 1998b), H. triquetrifolium (Roz et al., 2002) and H. caprifoliatum (Viana et al., 2005). In these reported study, several in vivo and in vitro studies have demonstrated that the antidepressant properties and the uptake inhibitory effects of the extracts obtained from these species were due to their main phloroglucinol derivatives such as hyperforin from H. perforatum and H. triquetrifolium (Chatterjee et al., 1998a,b; Mller et al., 1998; Jensen et al., 2001; Roz et al., 2002), and HCl from H. caprifoliatum (Viana et al., 2005). These reported studies have considered that hyperforin (Chatterjee et al., 1998b; Mller et al., 1998; Roz et al., 2002) and HCl (Viana et al., 2005) operate their uptake inhibitory effect in a nonspecific manner since they inhibit this uptake of each monoamine in a non-competitive manner. Thus, their mechanism of action does not seem to correspond to a direct interaction with the respective monoamine transporters. Taken together, the present study as well as those reported by other authors (Chatterjee et al 1998a,b; Mller, 2003; Roz and Rehavi, 2003; Viana et al., 2005) corroborate the idea that the

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mechanism of action for Hypericum extracts, likely differs from that of known antidepressants. In fact, all clinically available antidepressants directly interact with monoaminergic transmissions either by inhibitory enzymes of their catabolism, or blocking the transporters involved in their neuronal uptake, or by stimulation of receptors expressing their actions (Millan, 2004). It is commonly admitted that the increase of monoamine levels in the synaptic cleft induced by uptake inhibition, due to the blockade of plasma membrane transporters for monoamines, triggers the series of neurochemical modifications induced by antidepressants (Manji et al., 2001; Morilak and Frazer, 2004; Millan, 2004). The molecular mechanisms by which hyperfoliatin or other phloroglucinol derivatives induce antidepressantlike effect activity remain unclear. Recent reports, investigating H. caprifoliatum extract (Viana et al., 2005) or H. perforatum extract (Gobbi et al., 1999; Roz et al., 2002; Yoshitake et al., 2004), suggested the possibility that their effects could have resulted not from a modification of monoamine uptake transporters function, but more likely to an increase of the neurotransmitter levels in the synaptic cleft, due to a marked efflux of monoamine. In conclusion, the results obtained in our study are in accordance with the previous reported studies (Chatterjee et al., 1998a,b; Mller et al., 1998; Jensen et al., 2001; Roz et al., 2002; Viana et al., 2005) regarding the antidepressant-like effect of Hypericum phloroglucinol contents in the forced-swimming test, an animal model for selecting antidepressant drugs with a good predictive value (Porsolt et al., 1991). They further support recent studies suggesting that a mechanism of action of phloroglucinol derivatives could be related to the inhibition of neuronal monoamine uptake, probably not caused by a direct effect of hyperfoliatin on known binding sites on the transporters. Acknowledgements This work was supported by grants from the Centre National de la Recherche Scientifique (CNRS, France). The authors thank Richard Medeiros, Rouen University Hospital Medical Editor for his valuable advice in editing the manuscript. References
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